NOVEL LIPOLYTIC ENZYMES

Abstract: Novel lipolytic enzymes are disclosed which are capable of removing substantial amounts of lard from a lard stained swatch in a one cycle wash. Preferred lipolytic enzymes are variants of the Humicola lanuginosa lipase which may be prepared by recombinant DNA techniques. The enzymes are advantageously used in detergent compositions.

Description: NOVEL LIPOLYTJC ENZYMES FIELD OF THE INVENΗON The present invention relates to novel lipolytic enzymes which are capable of removing a substantial amount of fatty matter during a one cycle wash. BACKGROUND OF THE INVENΗON For a number of years lipolytic enzymes have been used as detergent enzymes, i.e. to remove lipid or fatty stains from clothes and other textiles. For instance, various microbial lipases have been suggested as detergent enzymes. Examples of such lipases include a Humicola lanuginosa lipase, e.g. described in EP 258 068 and EP 305 216, a Rhizomucor miehei lipase, e.g. as described in EP 238 023 and Boel, et al., Lipids 23, 701-706, 1988, , Absidia sp. lipolytic enzymes (WO 96/13578), a Candida lipase, such as a C antarctica lipase, e.g. the C. antarctica lipase A or B described in EP 214 761, a Pseudomonas lipase such as a Ps. alcaligenes and Ps. pseudoalcaligenes lipase, e.g. as described in EP 218 272, a Ps. cepacia lipase, e.g. as described in EP 331 376, a Pseudomonas sp. lipase as disclosed in WO95/14783, a Bacillus lipase, e.g. a B. subtilis lipase (Dartois et al., 1993), a B. stearothermophilus lipase (JP 64/744992) and a B. pumilus lipase (EP 91 00664). Furthermore, a number of cloned lipases have been described, including the Penicillium camembertii lipase described by Yamaguchi, S. et al., 1991, the Geot ή cum candidum lipase (Schimada, Y. et al., 1989, Schrag, J.D et al (1991) Nature 351, p. 761), and various Rhizopus lipases such as a R. delemar lipase (R.D. Joerger and MJ.Hass (1993), Lipids 28 p. 81-88), a R. niveus lipase (W. Kugimiya et al. (1992), Biosci. Biotech. Biochem. 5, p. 716- 719), R. javinicus (W. Uyttenbroeck et al. (1993) Biol. chem. Hoppe-Seyler 374, p. 245-254) and aR. oryzae (Haas, M.J., Allen, J. and Berka, T.R. (1991) Gene 109, p.107-113) which has a substantially identical sequence to the other Rhizopus lipases. Other types of lipolytic enzymes having been suggested as detergent enzymes include socalled cutinases, e.g. derived from Pseudomonas mendocina as described in WO 88/09367, or a cutinase derived from Fusarium solanipisi (e.g. described in WO 90/09446). In recent years attempts have been made to prepare lipase variants having improved properties for detergent purposes. For instance, WO 92/05249 discloses lipase variants with improved properties, in which certain characteristics of wild-type lipase enzymes have been changed by specific, i.e. site-directed modifications of their amino acid sequences. More specifically, lipase variants are described, in which one or more amino acid residues of the so¬ called lipid contact zone of the parent lipase has been modified. WO 94/01541 describes lipase variants with improved properties, in which an amino acid residue occupying a critical position vis a vis the active site of the lipase has been modified. EP 407225 discloses lipase variants with improved resistance towards proteolytic enzymes, which have been prepared by specifically defined amino acid modifications. EP 260 105 describes hydrolases in which an amino acid residue within 15 A from the active site has been substituted. WO 95/35381 discloses Pseudomonas sp. lipase variants, in particular P. glumae and P. pseudoalcaligenes lipase variants which have been modified so as to in¬ crease the hydrophobicity at the surface of the enzyme. WO 96/00292 discloses Pseudomonas sp. lipase variants, in particular P. glumae and P. pseudoalcaligenes lipase variants which have been modified so as to improve the en¬ zyme's compatibility to anionic surfactants, WO 95/30744 discloses mutant lipases such as Pseudomonas sp. lipases which have been modified to an increased surfactant resistance WO 94/25578 discloses mutant lipases comprising at least a substitution of the me¬ thionine corresponding to position 21 in the P. pseudoalcaligenes lipase, in particular to leucine, serine or alanine. All of the above mentioned lipase variants have been constructed by use of site-directed mutagenesis resulting in a modification of specific amino acid residues which have been chosen either on the basis of their type or on the basis of their location in the secondary or tertiary structure of the parent lipase. An altemative approach for constructing mutants or variants of a given protein has been based on random mutagenesis. For instance, US 4,898,331 and WO 93/01285 disclose such techniques. WO 95/22615 discloses variants of lipolytic enzymes having an improved washing performance, the variants having been prepared by a method involving subjecting a DNA sequence encoding the parent lipolytic enzyme to random mutagenesis and screening for variants having a decreased dependence to calcium and/or an improved tolerance towards a detergent or one or more detergent components as compared to the parent lipolytic enzyme. WO 95/09909 discloses, inter alia, chemically modified lipases or lipase mutants which has a higher pi than the corresponding modified enzyme. A drawback of all detergent lipolytic enzymes described until now is that they exert the best fat removing effect after more than one wash cycle, presumably because the known lipolytic enzymes, when deposited on the fatty stain to be removed, are more active during a certain period of the drying process than during the wash process itself (Gormsen et al., in Proceedings of the 3rd World Conference on Detergents, AOCS press, 1993, pp 198-203). This has the practical consequence that at least two wash cycles (separated by a sufficient drying period) are required to obtain a substantial removal of fatty stains. Some lipolytic enzymes have been described as allegedly being capable of removing fatty matter during the first wash cycle. Thus, WO 94/03578 discloses a detergent composition which in addition to various detergent components an enzyme which is alleged to be capable of exhibiting a substantial lipolytic activity during the main cycle of a wash process. Examples of lipolytic enzymes allegedly exhibiting the above activity include stem- specific cutinases such as the cutinase from Fusarium solanipisi, Fusarium roseum culmorum, Rhizoctonia solani and Alternaria brassicicola. However, when tested under realistic washing conditions none of these enzymes are capable of removing substantial amounts of a fatty stain during a one cycle wash process (cf the examples hereinafter). Thus, a need exists for lipolytic enzymes which under realistic wash conditions are capable of removing substantial amounts of fatty matter during one wash cycle. SUMMARY OF THE INVENTION The present inventors have now suφrisingly identified and constructed a novel class of lipolytic enzymes which are capable of removing substantial amounts of a fatty material during a one cycle wash performed under realistic washing conditions. The present invention is based on this finding. Accordingly, in a first aspect the invention relates to a lipolytic enzyme which, when present in detergent composition A and/or B defined herein, is capable of removing at least 15% more lard from a lard stained swatch than the same detergent composition without the enzyme, in a one cycle wash assay comprising subjecting 7 lard-stained cotton swatches (9 x 9 cm) per beaker to a one cycle wash in a thermostated Terg-O-to-Meter (TOM), each beaker containing 1000 ml of water comprising 3.2 mM Ca 2+ /Mg 2+ (in a ratio of 5: 1) and 5 g/1 of said detergent composition, pH 10, and comprising 12500 LU/1 of the lipolytic enzyme, the wash treatment being carried out for 20 minutes at a temperature of 30°C, followed by rinsing for 15 minutes in running tap water and ovemight linedrying at room temperature, subsequent extraction and quantification of fatty matter on the swatches by Soxhlet extraction. The Detergent Composition A and/or B and the one cycle wash assay are further described in the Materials and Methods section herein. The present invention constitutes the first true demonstration of the suφrising fact that it is possible to develop (identiiy and/or create) first wash lipolytic enzymes. Thus, what hitherto has been considered impossible (based on several years intensive research by a number of research teams throughout the world (as reflected by the number of hopeful patent applications filed in this field as mentioned above)) has now been shown to be possible. The present inventors have developed very convenient and succesful methods for creating first wash lipolytic enzymes. Accordingly, in a second important aspect the invention relates to a method of preparing a first wash mutated lipolytic enzyme, which method comprises at least the following steps: (a) subjecting a DNA sequence encoding a parent lipolytic enzyme to mutagenesis, conveniently random mutagenesis to form a variety of mutated DNA sequences; (b) expressing the mutated DNA sequences in host cells; (c) screening for host cells expressing a mutated lipolytic enzyme which has a decreased dependence on calcium and/or an improved tolerance towards a detergent or a detergent component as compared to the parent lipolytic enzyme; and (d) selecting a mutated lipolytic enzyme among those resulting from step (c) which, when present in the detergent composition A and or B in a concentration of 12500 LU/1, is capable of removing at least 15% more lard from a lard stained swatch, than the same detergent composition without the enzyme, in the one cycle wash assay described above. In a further aspect the invention relates to a method of preparing a first wash mutated lipolytic enzyme which method comprises at least the following steps: (a) constmcting mutated DNA sequences by combining a DNA sequence encoding a first parent lipolytic enzyme and a DNA sequence encoding a second parent lipolytic enzyme and optionally fiirther DNA sequences encoding a third (and optionally further) parent lipolytic enzymes, the DNA sequences being sufficiently homologous to allow for recombination between parts of or the entire DNA sequences to take place, (b) expressing the resulting mutated DNA sequences in host cells, and (c) selecting a mutated lipolytic enzyme encoded by a mutated DNA sequence which, when present in detergent composition A or B in a concentration of 12500 LU/1, is capable of removing at least 15 % more lard from a lard stained swatch than the same detergent composition without the enzyme, in the one cycle wash described above. In a prefened embodiment the methods according to the second and third aspects of the invention are combined, i.e. a mutated lipolytic enzyme resulting from the method of the second aspect is used as a parent enzyme in the method according to the third aspect. In a further aspect the invention relates to a DNA constmct comprising a DNA sequence encoding a first wash lipolytic enzyme as defined above. In a still further aspect the invention relates to a recombinant expression vector carrying the DNA constmct, a cell which is transformed with the DNA constmct or the vector as well as a method of producing a first wash lipolytic enzyme by culturing said cell under conditions conducive to the production of the enzyme, after which the enzyme is recovered from the culture. In final aspects the invention relates to the use of a first wash lipolytic enzyme as a detergent enzyme, in particular for washing or dishwashing, and to a detergent additive and a detergent composition comprising the enzyme. BRIEF DESCRIPTION OF THE DRAWING Fig.1 shows the plasmid pYESHL, Fig. 2 the plasmid pAOl, Fig. 3 the plasmid pAHL, Figs 4 and 5 a graphical illustration of a PCR mutagenesis method, Fig. 6 the plasmid pJSO37; Fig. 7 the constmction of the plasmid pDM177, Figure 8 the constmction of Aspergillus vector pCaHj485 Figure 9 shows the original sequence of the Absidia reflexa ATTC 44896 lipase. The trip¬ led; coding for the first amino acid serine of the mature NL127 as well as the stop codon are underlined. Figure 10 shows the Absidia reflexa ATTC 44896 sequence in the context of the yeast expression vector pTiK05. Figure 11 shows the mating factor αl signal sequence DEFINITIONS AND BACKGROUND ON LIPASE STRUCTURE Definitions of terms used in the present application In the present context the term "lipolytic enzyme" is intended to indicate an enzyme classified under the Enzyme Classification number E.C. 3.1.1 (Carboxylic Ester Hydrolases) in accordance with the Recommendations (1992) of the Intemational Union of Biochemistry and Molecular Biology (IUBMB)). Lipolytic enzymes thus exhibit hydrolytic activity towards at least one of the types of ester bonds present in at least one of the following lipids: mono-, di- and triglycerides, phospholipids (all classes), thioesters, cholesterol esters, wax-esters, cutin, suberin, synthetic esters, etc. (cf. the different types of esterbonds mentioned in the context of E.C. 3.1.1). Thus, the lipolytic enzyme may, e.g., be what has conventionally been termed a lipase, a phospholipase, an esterase or a cutinase. The term "lipolytic enzyme" is intended to embrace naturally-occurring enzymes as well as enzymes, which as compared to a naturally-occurring enzyme, have been modified, e.g. by modification of one or more amino acid residues of the enzyme or by chemical modification. In the present context the latter type of a lipolytic enzyme is termed a variant. In the present context the capability of the enzyme in removing a substantial amount of fatty matter during a one cycle wash is also refened to as a first wash effect, a one cycle wash effect, a through-the-wash effect, and the like. Analogously, lipolytic enzymes of the invention, which are capable of effecting removal of a substantial amount of a fatty material during a one cycle wash, are called first wash lipolytic enzymes, through-the-wash lipolytic enzymes, one cycle wash lipolytic enzymes, and the like. In the present context the term "Detergent Composition A and/or B" as used to define the lard removing capability of a given lipolytic enzyme of the invention is intended to indicate that the lipolytic enzyme has the indicated lard removing capability when present in either or both of Detergent Compositions A and B. In the present context the term "parent enzyme" is intended to include a naturally- occurring enzyme as well as a variant of such enzyme. Accordingly, the term is used to identify the starting material to be modified in accordance with a method of the invention for preparing first wash lipolytic enzymes, irrespectively of whether said starting material is a naturaUy-occurring enzyme or a variant of such enzyme. The term "a variety of mutated sequences" as used about the method according to the second aspect is intended to be understood to indicate that at least two, but preferably a much higher number of different sequences, such as at least 10, at least 50 at least 100, at least 1000 sequences have resulted from the mutagenesis. The term "random mutagenesis" is intended to be understood in a conventional manner, i.e. to indicate an introduction of one or more mutations at random positions of the parent enzyme or introduction of random amino acid residues in selected positions or regions of the parent enzyme. The random mutagenesis is normally accompanied by a screening which allows the selection of mutated lipolytic enzymes which, as compared with the parent enzyme, have improved properties. Suitable techniques for introducing random mutations and screening for improved properties are discussed in detail below. The term "satisfactory wash performance" as used about lipolytic enzymes disclosed herein is intended to indicate that the enzyme has an improved performance when tested in a suitable wash assay or a wash related assay (such as the assays described in the Materials and Methods and Example 5 below) as compared to the commercially available lipolytic enzymes (Lumafast and Lipomax from Genencor, Lipolase and Lipolase Ultra from Novo Nordisk and Liposam (from Showa Denko). The improved performance may be in terms of lipid stain removing capability and/or a decreased calcium dependency, an improved tolerance towards a detergent or detergent component, an increased hydrophobicity, an interesting substrate specificity, or the like. In the present context, the term "decreased dependence on calcium" as used in connection with the screening for mutated lipolytic enzymes, in particular lipolytic enzymes exhibiting enzymatic activity towards lipase substrates having hydrocarbon chains (ffa-part) of a length exceeding approx. 6-8 C-atomes, is intended to mean that the mutated lipolytic enzyme requires lower amounts of Ca 2+ for exhibiting the same degree of activity and/or stability as the parent enzyme when tested under similar conditions. In other words the stability and/or activity of the enzyme is/are increased in the absence of calcium as compared to that of the parent enzyme. The stability may, e.g. , be assayed by a determination of residual activity upon preincubation under Ca-free conditions and/or DSC (Differential Scanning Calorimetry) in the absence/presence of free Ca2+. Preferably, the mutated lipolytic enzyme of the invention is substantially independent of the presence of calcium for exhibiting enzymatic activity, in particular at a pH higher than 8. The term "improved tolerance towards a detergent or detergent component" as used in connection with the screening for mutated lipolytic enzymes is intended to mean that the mutated lipolytic enzyme is active at higher concentrations of the detergent or detergent component than the parent lipolytic enzyme. In the present context the term "detergent" is intended to indicate a mixture of detergent ingredients normally used for washing or dishwashing. Analogously, a "detergent component" is intended to indicate a component or ingredient normally found in detergent or dishwashing compositions, specific examples of which are given in the section further below entitled "Detergent compositions". Background on lipolytic enzyme structure and definition of structure terminology The 3D stmcture of a number of lipolytic enzymes has been determined. It has been found that the stmctures have a common motif in the core of the protein consisting of a central β-sheet, one of the strands ending in a nucleophil elbow including the active serine residue (Ollis et al, 1992). Lipolytic enzymes comprise a lipid contact zone which is a surface with increased surface hydrophobicity which interacts with the lipid substrate at or during hydrolysis. For lipolytic enzymes containing a lid the lipid contact zone is typically formed when the enzyme is activated by substrate (and the lid thereby displaced). For lipolytic enzymes which do not contain a lid there is generally little or no conesponding substantial movement leading to the creation of the lipid contact zone. The lipid substrate is a conglomerate of single lipid substrate molecules. The lipid contact zone contains a binding area to which a single lipid substrate molecule binds before hydrolysis. This binding area contains an acyl-binding hydrophobic cleft and a so-called hydrolysis pocket, which is situated around the active site Ser, and in which the hydrolysis of the lipid substrate is believed to take place. The lipid contact zone includes one or more protein secondary stmcture elements, i.e. , loop sequences, the amino acid residues of which contact, bind to and/or interact with the substrate during hydrolysis when the lipolytic enzyme is activated. The lipid contact zone may be recognized, e.g. from a three-dimensional stmcture of the lipolytic enzyme in question created by suitable computer programs. The lipid contact zone may be identified by searching the stmcture for the relevant features defining the zone, including a zone positioned on top of the active site residues and containing a lid stmcture (for lipolytic enzymes containing a lid) which when opened creates a hydrophobic surface containing a narrow hydrophobic binding pocketThe conformation of the inactive and activated H. lanuginosa lipolytic enzyme, respectively, is shown in Figs. 1 and 2 of WO 5 92/05249. In terms of amino acid residues the lipid contact zone of the H. lanuginosa lipolytic enzyme is defined by amino acid residues 21-25, 36-38, 56-62, 81-98, 110-116, 144-147, 172-174, 199-213 and 248-269. These residues have been identified on the basis of computer model simulations of the interaction between the lipolytic enzyme and a lipid substrate. For 10 lipolytic enzymes having substantially the same stmcture as the H. lanuginosa lipolytic enzyme, e.g. the lipolytic enzymes produced by Rhizomucor miehei, by the Rhizopus oryzae, by Penicillium camembertii and by Absidia sp. (cf the "Background of the Invention" section above) the lipid contact zone is constituted by amino acid residues occupying homologous positions to those given above for the H. lanuginosa enzyme. The homologous positions may 15 be identified by an alignment of the relevant amino acid sequences (e.g. using the UWGCG GAP programme) looking for groups of sequence similarity, but may more conveniently be done by comparing the stmctures or stmcture models of the relevant enzymes. More specifically, the lipid contact zone of these enzymes is constituted of the following residues (the numbering used refers to the amino acid residue in the mature enzyme, the sequence of 20 which is apparent from the references disclosed in the Background of the Invention section above unless otherwise indicated): Penicillium camembertii: 21-25, 36-38, 56-62, 81-98, 109-115, 143-146, 172-174, 198-212, 247-280; Rhizopus oryzae: 29-33, 39-41, 57-62, 81-98, 109-115, 143-146, 175-177, 202-216, 25 245-269. Rhizomucor miehei: 29-33, 39-41, 56-61, 80-97, 108-114, 142-145, 174-176, 201-215, 245- 269; Absidia sp. lipase: 29-33, 39-41, 56-61, 80-97, 108-114, 142-145, 171-173, 198-212, and 239-263, the numbering based on that the mature enzyme has the following N-terminal 30 sequence: SSKQDYR. The entire sequence is apparent from (SEQ ID NO 98). As an altemative or in addition to the homology based identification of the lipid contact zone, the lipid contact zone may be identified by a) calculating the hydrophobic vector of the 3-D molecular stmcture of the activated enzyme; b) making a cut perpendicular to the vector through the CA-atom (Cα-atom) of the second amino acid residue after the active site serine in the linear sequence; c) including all residues with at least one atom on that side of the cut to which the vector 5 points; and d) selecting from those residues, those which have at least one atom within 5 Angstrom of the surface of the protein. The hydrophobic vector is calculated from the protein stmcture by summing up all residue vectors for residues having a surface accessibility (Lee et al., Mol. Biol. 55, pp. 379- 10 400 (1971)) of at least 15 % . The starting point of the residue vector is defined as the CA- atom of the residue and its direction is through the mass centre of the sidechain. The magnitude of each residue vector is defined as the residues relative free energy of transfer between water and a more hydrophobic solvent (see, e.g., Creighton, Protein, W. Freeman & Co., p. 151 (1984)). The surface accessibility of each residue is calculated using the Connolly 15 program (Lee et al., op. cit.). Using the above method and/or the alignment of the various sequences, which is apparent from Svendsen et al, Biochimica et Biophysica Acta, 1259 (1995) 9-17, the following lipid contact zones of lipolytic enzymes isolated from various Pseudomonas sp. have been identified (the numbering used refers to the amino acid residues of the mature 0 enzyme as presented in the above mentioned publication (Svendsen et al. (1995))): Pseudomonas cepacia lipase: 15-36, 110-167, 209-266, 281-304; Pseudomonas pseudoalcaligenes lipase: 15-35, 106-163, 200-232, 250-271; Pseudomonas glumae: 15-36, 110-167, 209-266, 281-304; Pseudomonas mendocina (SD702) lipase: 19-39, 111-166, 213-244, 258-279 (the sequence is 5 apparent from WO 95/14783), Pseudomonas s/?.(Liposam ® ) lipase: 17-37, 109-161, 208-239, 253-274 (SEQ ID NO 99). Pseudomonas wisconsinensis lipase: 13-34, 106-161, 200-242, 250-270 (the sequence is apparent from WO96/12012) The lipid contact zone for lipolytic enzymes which do not contain a lid structure may be 0 determined from the topology of the core as evaluated in a stmcture or model of the three- dimensional stmcture of the lipolytic enzyme. In this manner the lipid contact zone of the Fusarium solanipisi lipolytic enzyme has been determined to amino acid residues 40-50, 78- 91, 119-121, 147-154, 171-193 (as evaluated on the basis of the mature enzyme). Some lipolytic enzymes also comprise a surface loop stmcture, i.e. , a lid, which is part of the lipid contact zone. The surface loop stmcture covers the active serine when the lipolytic enzyme is in inactive form. When the enzyme is activated, the surface loop stmcture shifts to expose the active serine residue. The surface loop stmcture has a predominantly hydrophobic inner surface facing the binding pocket and a predominantly hydrophilic outer surface. Example of lipolytic enzymes which have a surface loop stmcture are those produced by Humicola lanuginosa, Rhizjomucor miehei, Rhizopus sp., Penicillium camembertii and Absidia sp., a number of Pseudomonas sp., such as Ps. cepacia, Ps. aeroginosa, Ps. fragi (cf. the "Background of the Invention" section above), Candida rugosa (Grochulski.P et al (1993) J. Biol. Chem. 268, p. 12843) and the human pancreatic lipase described in Winkler et al., Nature 343, pp. 771-74 (1990). The surface loop stmcture of the lipolytic enzyme produced by Humicola lanuginosa DSM 4109 is defined by amino acid residues at positions 82-96. The surface loop stmcture of lipolytic enzymes with substantially the same three-dimensional stmcture (cf above) is defined by the amino acid residues occupying homologous positions to those of the H. lanuginosa lipolytic enzyme, i.e.81-98 (for the Penicillium camembertii lipase), 82-99 (for Rhizopus oryzae), 80-97 (for Rhizomucor miehei), 80-97 (foτ Absidae sp lipase). The surface loop stmcture of a representative number of lipolytic enzymes produced by Pseudomonas sp. are: Ps. glumae: 135-155, Ps. cepacia 135-155, Ps pseudoalcaligenes 132- 152), Pseudomonas sp lipase (SD705) (Liposam ® ) 129-149. shown in SEQ ID NO 99. DETAILED DESCRIPTION OF THE INV-ENΗON The first wash lipolytic enzyme of the invention Preferably, the lipolytic enzyme of the invention is capable of effecting an even higher lard removing capability than that stated above. Accordingly, in a prefened embodiment Detergent Composition A and/or B comprising the lipolytic enzyme of the invention is capable of removing at least 15%, such as at least 20% more lard, than Detergent Composition A and/or B, respectively, which does not comprise the lipolytic enzyme, when tested in the one cycle wash assay described herein in a concentration of 12500 LU/1. In a more preferred embodiment the lipolytic enzyme is one, which, when present in Detergent Composition A and/or B allows the detergent composition to remove at least 25% such as at least 30% or 35% or 40% or 50% more lard than Detergent Composition A and/or B without the lipolytic enzyme, when tested in the one cycle wash assay as described herein. The concentration of lipolytic enzyme used in the above described one cycle wash assay (i.e. 12500 LU/1) may be considered high for practical applications, but has been chosen for assay purposes in order to minimize the analytical variation. A more realistic concentration is 1250 LU/1 which in an altemative embodiment may be used to define the lard removing capability of a lipolytic enzyme of the invention. Accordingly, in a further embodiment the lipolytic enzyme is one which is capable of removing at least 15 % , such as at least 20% more lard, than Detergent Composition A and or Detergent Composition B which does not comprise the lipolytic enzyme, when used in the one cycle wash assay described herein in a concentration of 1250 LU/1. In an even more preferred embodiment the lipolytic enzyme, when present in Detergent Composition A and/or B in a concentration of 1250 LU/1, allows the detergent composition to remove at least 25% such as at least 30% or 35% more lard than Detergent Composition A and/or B without the lipolytic enzyme, when used in a one cycle wash assay as described herein. In prefened embodiments the first wash lipolytic enzyme of the invention is capable of removing: ■ when present in Detergent composition A in a concentration of 1250 LU/1 at least 15 % more lard from a lard stained swatch than Detergent composition A without the enzyme, ■ when present in Detergent A in a concentration of 12500 LU/1 at least 40% more lard from a lard stained swatchthan Detergent Composition A without the enzyme, ■ when present in Detergent composition B in a concentration of 1250 LU/1 at least 15% more lard from a lard stained swatch than Detergent composition B without the enzyme, ■ when present in Detergent B in a concentration of 12500 LU/1 at least 15% more lard from a lard stained swatchthan Detergent Composition B without the enzyme, when tested in a one cycle wash assay as described herein. In Example 5 herein a comparison is shown between the fat removing capability of lipolytic enzymes of the invention and that of lipolytic enzymes described in WO 94/03578 alleged to have a through-the-wash-effect. It is seen that the enzymes of the invention removed substantially more lard in a one cycle wash than the prior art enzymes. The comparison between the enzymes has been done by use of the same assay. While the lipolytic enzyme of the invention may be of any of the above mentioned types of lipolytic enzymes such as a hydrolase exhibiting activity towards ester and/or phospholipid bonds, it is particularly prefened that the enzyme is a lipolytic enzyme which exhibits activity towards esterbonds in mono-, di- and/or tri-glycerides and/or which exhibits activity towards cutin. Such enzymes are generally considered to be of high interest as detergent enzymes. In the Materials and Methods section and in Example 5 below suitable assays for identifying first wash lipolytic enzymes are given. These assays may be used to identify naturally-occurring first wash lipolytic enzymes. More specifically, in order to identify a naturally-occurring first wash lipolytic enzyme according to the invention candidate enzymes are recovered from suitable organisms expected to produce lipolytic enzymes, such as organisms which are taxonomically related to the ones given in the "Background of the Invention" section above or discussed later on in the "Parent Lipolytic Enzymes"section, or organisms which are found in an environment which require the organism to produce lipolytic enzymes in order to prevail. Subsequently, the recovered enzymes are subjected to the first wash lipolytic enzyme assays disclosed herein. Although the first wash lipolytic enzyme of the invention may be a novel naturally- occurring enzyme (identified on the basis of its first wash performance) it is presently preferred that the enzyme is a modified enzyme, i.e an enzyme which has been prepared by subjecting a parent lipolytic enzyme to mutagenesis and/or to chemical modification so as to result in a modified lipolytic enzyme which has a first wash activity. The parent lipolytic enzyme may be one which has a first wash activity (which may thus be improved by the mutagenesis or chemical modification) or may be without any first wash activity as defined herein. In one embodiment it is considered advantageous that the parent enzyme has a satisfactory wash performance itself or even a first wash performance, the latter property then being improved by the mutation(s). Parent enzymes with a satisfactory (but not necessarily a first wash performance) may be selected using the assay described in Example 6 hereinafter. The chemical modification of amino acid residues of the parent enzyme may e.g. be performed in accordance with the principles disclosed in WO 95/09909 the content of which is incorporated herein by reference. For instance, the chemical modification may be accomplished by coupling an amine ligand (such as an aminated sugar, aminated alcohol or EET aminated glucosamine or isomeric forms thereof) to the carboxyl group of glutamic acid or aspartic acid residues in the enzyme. The chemical modification may be performed by methods known in the art, such as those described in WO 95/09909. The chemical modification may be done on acid groups so as to remove negative charges. The mutagenesis of the parent lipolytic enzyme is preferably done so as to improve the substrate binding affinity of the parent enzyme. More specifically, it has been found that an improved substrate binding affinity may result in a first wash activity being obtained. It is presently contemplated that an improved substrate binding affinity may be achieved by making the surface of the parent enzyme less negative. Accordingly, the mutagenesis may be performed so as to replace at least one neutral amino acid residue located at the surface of the parent enzyme with a positively charged amino acid residue, deleting a negatively charged amino acid residue located at the surface of the parent enzyme or replacing a negatively charged amino acid residue located at the surface of the parent enzyme with a neutral (including hydrophobic) or positively charged amino acid residue. Amino acid residues located at the surface of the enzyme may be identified by use of the Conolly program refened to in the Definitions section above. In a prefened embodiment the mutagenesis is performed so as to remove the amino acid residue D and/or E, and/or to insert, conveniently by replacement, of R, K, W, F, Y, I, L. A suitable test for an improved substrate binding affinity is described in example 11 hereinafter. The 1st wash effect of the above changes from negative towards positive surface may be improved and/or stabilized by introduction of exchanges optimizing the stmcture or stability. Thus, for instance introduction of a proline residue into the enzyme surface may lead to an increased proteolytic and/or thermal stability; introduction of hydrophilic amino acid residues, e.g. Glu and/or Asp, may increase the anionic detergent stability, and the introduction of hydrophobic amino acid residues may increase the adrøφtioiVaffinity of the enzyme. The introduction of the above type of amino acid residues may either be accomplished by simply inserting the amino acid residues into a suitable location at the surface of the enzyme or by replacing amino acid residue(s) located at such position(s). It is presently believed that a first wash lipolytic enzyme of the invention is a variant of a parent lipolytic enzyme which comprises at least one mutation, but typically more mutations, preferably located at the surface of the enzyme. The variant may comprise more mutations such as at least 2, 3, 4 or 5 mutations, e.g. in the range of 1-20, 1-15, 1-12, 1-10, 1-9, 1-8, 1-7, 1-6, 1-5 or 1-4 mutations, or any number of mutations which does not impair the enzymatic activity of the enzyme. It has been found that mutations within as well as outside the lipid contact zone of the parent H. lanuginosa lipase disclosed herein may be of importance for achieving a first wash activity. Accordingly, the first wash lipolytic enzyme of the invention carrying a mutation may be constmcted from a parent lipolytic enzyme by modification of at least one amino acid residue outside the lipid contact zone of the parent enzyme and/or by addition of at least one amino acid residue outside said zone, and/or by modification of at least one amino acid residue within the lipid contact zone of the parent enzyme and/or by addition of at least one amino acid residue within said zone. Accordingly, in another embodiment the lipolytic enzyme of the invention is one, which has been prepared from the parent enzyme by modification, deletion or substitution of at least one amino acid residue in the lipid contact zone of the parent enzyme or addition of at least one amino acid residue to said zone. In a still further embodiment the lipolytic enzyme is one which has been prepared from the parent enzyme by modification, deletion or substituion of at least one amino acid residue outside the Upid contact zone or addition of at least one amino acid residue to said zone, the amino acid residue preferably being located at the surface of the parent enzyme. The mutations within or outside the lipid contact zone are preferably conducted to as to improve the substrate binding affinity of the resulting modified enzyme, conveniently be removal of negative charges as described above. Although site-directed mutagenesis following the above principles (and combined with testing of the resulting enzyme variants for first wash activity) may be used for the creation of first wash lipolytic enzymes it is presently prefened to use other methods of creating first wash lipolytic enzymes. Random mutagenesis, in particular localized random mutagenesis, as well as in vivo recombination of homologous genes have been found to be of particular interest for that purpose - these methods are described in detail further below. First wash lipolytic enzyme modified in a non-structural part of its C- or N-terminus It has suφrisingly been found that it is possible to confer a first wash effect to a parent lipolytic enzyme or to significantly enhance the first wash effect of a parent lipolytic enzyme by applying at least one N-terminal and/or C-terminal peptide addition at or within a non- stmctural part of the parent enzyme in its mature form or by introducing other changes in a non-stmctural part of the C-terminal and/or N-terminal end of the parent mature enzyme. Accordingly, in a further highly prefened embodiment the first wash lipolytic enzyme of the invention is a variant of a parent lipolytic enzyme which, as compared to the parent enzyme, has been modified at or within a non-stmctural part of the N- and/or C-terminal end of the parent enzyme. In the present context the term "modified" is intended to indicate that i) an appropriate peptide addition has been applied to the parent enzyme or ii) one or more amino acid residues within the non-stmctural part of the C-terminal and/or N-terminal part of the parent mature enzyme has/have been deleted or replaced by other amino acid residues, or iii) the parent enzyme has been modified by a combination of i) or ii). In the present context first wash lipolytic enzymes of the invention which have been modified in this way may be termed a modified enzyme of the invention. In the present context the term "peptide addition" is intended to indicate that a stretch of one or more consecutive amino acid residues has been added to either or both of the N- and/or C-terminal end(s) of the parent enzyme (i.e. fused to the first and/or last amino acid residue ofthe parent enzyme) or inserted within the non-stmctural part of the N- and/or C-terminal end(s) of the parent enzyme. The modified enzyme may comprise a peptide addition at either the N-terminal or the C-terminal end or both in the N- and the C-terminal ends of the parent lipolytic enzyme. If a peptide addition is applied to both the N- and the C-terminus of the parent enzyme, the peptide additon at either terminus may have the same amino acid sequence or different amino acid sequence. Multiple copies of the same or different peptide additions may be inserted or added. The term "an appropriate peptide addition" is used to indicate that the peptide addition to be used is one which is capable of effecting a first wash performance or an improved first wash performance. The "appropriateness" of the peptide addition may be checked by a com¬ parative analysis of the first wash performance of a modified enzyme to which the peptide ad¬ dition has been applied and of the conesponding parent enzyme, respectively. The wash per¬ formance may, e.g., be determined by the one cycle wash assay described in the Materials and Methods section. It is presently contemplated that the improved first wash performance effected by the peptide addition is, at least in part, due to an increased affinity of the modified lipolytic en- zyme towards its lipid substrate (although this may not be the only reason). Accordingly, in a prefened embodiment the peptide addition is one which confer an increased affinity of the modified enzyme towards its lipid substrate. The term "mature enzyme" is used in its conventional meaning, i.e. to indicate the active form of the enzyme resulting after expression and posttranslational processing (to remove pro and/or pre-sequences) by the producer organism in question. When the enzyme is a secreted enzyme, the mature enzyme will normally be the form of the enzyme resulting after secretion. More specifically this means that the pre- and pro-peptide sequences, if present, have been removed from the initially translated enzyme, i.e. the unprocessed enzyme The term "non-stmctural part" is intended to indicate the part of the N- and C-terminal end, respectively, which is outside the first or last, respectively, stmctural element, such as an α-helix or β-sheet stmcture, of the folded mature enzyme. The non-stmctural part may easily be identified in a three-dimensional stmcture or model of the enzyme in question. Typically, the non-stmctural part comprises the first or the last about 1-20 amino acid residues of the amino acid sequence constituting the enzyme. For enzymes having a similar three-dimensional stmcture to that of the H. lanuginosa lipolytic enzyme the insertion may be made in the part of said enzyme which conesponds to a "non-stmctural part" of the H. lanuginosa lipolytic enzyme. The non-stmctural part of the H. lanuginosa lipolytic enzyme normally comprises the first or the last about 1-20 amino acid residues of the mature enzyme. It is presently believed that the capability of the peptide addition of providing the desired first wash effect depends on, e.g., the identity of the parent enzyme to be modified, the stmcture (including length) of the peptide addition, the impact of the peptide addition on the stmcture of the entire lipolytic enzyme, the nature or functionality of amino acid residues of the peptide addition, etc. A prerequisite for the peptide addition being capable of providing the desired effect is, of course, that the modified enzyme containing the peptide addition is expressible in a suitable host organism. The following general considerations may be of rele¬ vance for the design of a suitable peptide addition: Length of peptide addition: It has been found that peptide additions containing varying numbers of amino acid residues are capable of providing the desired first wash effect and thus, it is not possible to specify an exact number of amino acid residues to be present in the peptide addition to be used in accordance with the present invention. It is contemplated that EET the upper limit of the number of amino acid residues is determined, inter alia, on the impact of the peptide addition on the expression, the stmcture and/or the activity of the resulting modified enzyme. It is believed that the peptide addition may comprise a substantial number of amino acid residues, however, without all of these amino acid residues need to contribute to the desired first wash effect (even if the peptide addition contains a substantial number of amino acid residues only a small number of these need to provide the desired function, this small number may be termed the functional part of the peptide addition). The main considera¬ tion in relation to the lower Umit of the number of amino acid residues of the peptide addition will normaUy be that the number should be sufficient to provide the desired first wash effect. The peptide addition may thus comprise a single amino acid residue or an amino acid chain of from 2 and 500 amino acids, such as from 1 to 200, or from 2 to 100, preferably from 2 to 50, such as 3 to 50, even more preferably from 7-45 and still more preferably be¬ tween 1 and 15, such as between 1 and 10 or 1 and 7, especially between 4 and 10, such as 4 and 7 amino acids. StabiUty: The peptide addition should preferably be chosen so as to provide a modified Upolytic enzyme with a stable peptide addition and an acceptable stmctural stabiUty of the par¬ ent enzyme. For instance, a peptide addition which in itself forms a stmctural element, such as an α-helix or a β-sheet, may stabilize the resulting modified enzyme and thus be used in the context of the present invention. Peptide sequences capable of forming such stmctures are known in the art. Altematively, an improved stmctural stability may be provided by introduc¬ tion of cystein bridges in the modified Upolytic enzyme of the invention. For instance, a cys¬ tein bridge between the peptide addition and the mature part of the enzyme may be estabUshed if at least one of the amino acid residues of the peptide addition is a cystein residue which is located so as to be able to form a covalent binding to a cystein residue in the mature part of the enzyme. If no suitable cystein is present in the mature enzyme, a cystein may be inserted at a suitable location of said parent enzyme, conveniently by replacing an amino acid of the parent enzyme, which is considered unimportant for the activity. In addition, it may be desirable that at least one of the amino acid residues of the peptide addition is chosen so as to make the peptide addition less susceptibiUty to proteolytic degrada- tion by proteolytic enzymes of the host ceU used for expressing the modified Upolytic enzyme. For instance, the peptide addition may comprise at least one, and preferably at least two proline residues. Preferably, the peptide addition comprises 1-5, such as 1-4 or 1-3 or two or one proline residues. The proline residue(s) is(are) preferably placed at the proteolytic cleav¬ age site or close thereto. Altematively, the peptide addition may be one which provides a protease stable loop to the modified Upase, e.g. as described in EP 407225 or WO 93/11254. Nature of amino acid residues of the peptide addition: As stated above and without being Umited to any theory, it is presently beUeved that the improved performance may at least partly be due to an increased affinity of the modified Upolytic enzyme toward the substrate provided by the peptide addition. In particular, it is beUeved that favourable electrostatic inter¬ actions may be obtained between the negatively charged Upid surface and positively charged and/or hydrophobic amino acid residues present in the modified enzyme. Accordingly, it is particularly prefened that the modified enzyme of the invention comprises a peptide addition with at least one positive charge, such as at least 2, 3, 4 or more positive charges or expressed differently, in which a substantial number of the amino acid residues of the peptide addition is positively charged and/or hydrophobic. Analogously, and in order to reduce the negative charge in a non-stmctural end of the parent enzyme it is prefened to remove at least one such as two or more negatively charged amino acid residues from a non-stmctural N-terminal or C-terminal part of the parent enzyme of choice, in particular from the part of the parent Upase being constmcted of the 1-5 first or last N-terminal or C-terminal amino acid residues, such as 1-4, or 1-3 or 1-2. The negatively charged amino acid residue may either be removed or replaced by a neutral, a positively charged or a hydrophobic amino acid residue. For instance, the negatively charged amino acid residue to be removed may be an E or D which may be replaced with either of the positively charged amino acid residues R, K or H, the neutral amino acid residues S, T, G or Q, or the hydrophobic amino acid residues A, I, W F or L. Similarly, a neutral amino acid residue of a non-stmctural N-terminal or C-terminal part of the parent enzyme may be replaced with a positively charged or hydrophobic amino acid residue as defined above. Accordingly, the modified Upolytic enzyme of the invention in addition or as an altema¬ tive to a N-terminal and/or C-terminal extension may comprise a mutation in the non- stmctural C-terminal and/or N-terminal end of the parent enzyme, which mutation has in¬ volved deleting or replacing a negatively charged amino acid residue of said non-stmctural part with a positively charged or neutral amino acid residue or with a hydrophobic amino acid residue. If a peptide addition is present in both the N- and the C-terminal of the parent enzyme, the peptide addition at or within each of the terminals may have the same or a different amino acid sequence. Test of suitabiUty of peptide addition: the effect of using a given peptide addition, e.g. , designed on the basis of the above principles may be tested by constmcting a modified Upolytic enzyme containing the peptide addition and testing the properties of the resulting en¬ zyme for first wash activity. The peptide addition can be generaUsed in the foUowing way. The first residue (counted from the outer residue) is named "a", the second is named "b", the third "c" etc. Thus, in case of an N-terminal addition the first amino acid residue is termed "a", in case of a C-terminal addition the last amino acid residue is termed "a". In an important embodiment of the invention the peptide addition consists of from 1 to 7 amino acids. Such peptide addition, which can be appUed to both the N- and/or C-terminal of the parent enzyme, can be refened to as: a (one amino acid peptide addition) a-b (two amino acids peptide addition) a-b-c (three amino acids peptide addition) a-b-c-d (four amino acids peptide addition) a-b-c-d-e (five amino acids peptide addition) a-b-c-d-e-f (six amino acids peptide addition) a-b-c-d-e-f-g (seven amino acids peptide addition) Each letter defines an amino acid residue. a, b, c, d, e, f and g may independently be any amino acid including Alanine (A), Valine (V), Leucine (L), Isoleucine (I), Proline (P), Phenylalanine (F), Tryptophan (W), Methionine (M), Glycine (G), Serine (S), Threonine (T), Cysteine (C), Tyrosine (Y), Asparagine (N), Glutamine (Q), Aspartic acid (D), Glutamic acid (E), Lysine (K), Arginine (R), and Histidine (H). In specific embodiments a, b, c, d, e, f, and g are independently one of the foUowing amino acids: a: Leu, He, Val, Tφ, Phe, Ser, Arg, Cys, or Lys, b: Leu, lie, Val, Tφ, Phe Ser, Pro, Arg, Lys, Cys or His, c: Leu, lie, Val, Tφ, Phe, Ser, Pro, Arg, Cys, or Lys. d: Leu, lie, Val, Tφ, Phe Ser, Pro, Arg, Cys, or Lys. e: Leu, Be, Val, Tφ, Phe, Pro, Arg, Lys, Ala, Glu, Cys, or Asp, f: Leu, He, Val, Tφ, Phe, Pro, Arg, Lys, Ala, Glu, Cys, or Asp, g: Leu, -Qe, Val, Tφ, Phe, Pro, Arg, Lys, Cys, or Met. In a prefened embodiment at least one such as one, two, three or four of a, b, c, d, e, f, or g is a positively charged amino acid, i.e. Arg (R) or Lys (K) or a hydrophobic amino acid, i.e. Leu, Be, Val, Tφ or Phe. As stated further above, and dependent on the host ceU of choice it is generaUy beUeved that it is important that the peptide addition comprises at least one proline residue in order to protect the modified Upolytic enzyme against proteolytic degradation during the processing of the enzyme by the host ceU of choice. It may be desirable that the proline residue occupies position two (i.e. b) and/or three (i.e. c) of the peptide addition or a position close to the de¬ sired cleavage point (i.e. the point where processing by the host ceU in question is beUeved to occur). Accordingly, in one embodiment b and optionaUy c of the peptide addition is Pro In another embodiment of the invention a-b is SP (Ser-Pro), A-P or Q-P. If the peptide addition contains more amino acid residues, e.g. between 4 and 7 amino acids the peptide ad¬ dition has the general formula SPcd, SPcde, SPcdef, SPcdefg or APcd, APcde, APcdef, Ap- cdefg or QPcd, QPcde, QPcdef, QPcdefg. In each of these formulae c, d, e , f, and g may be any amino acid. However, preferred are the above mentioned group of amino acids. In another embodiment a-b comprise at least one positive amino acids (i.e. Arg and Lys) or hydrophobic amino acid residue (i.e. Leu, He, Val, Tφ and Phe). SpecificaUy, the peptide addition appUed to the parent Upolytic enzyme may advanta¬ geously be one of the foUowing amino acid residues or peptides: Arg (R), or Lys (K), or Leu (L), or Ile (I), or Val (V), or Tφ (W) or Phe (F), or Arg-Pro (RP), or Lys-Lys (KK), or Arg-Lys (RK), or Lys-Arg (KR), or Arg-Arg (RR), or Arg-Arg-Pro (RRP), or Arg-Pro- Val-Ser-Gln-Asp (RPVSQD) Ser-Pro-fle-Arg-Met (SPIRM), or Ser-Pro-Ile-Arg-Ala-Arg (SPIRAR), or Ser-Pro-He-Arg-Pro-Arg (SPIRPR) or Ser-Pro-De-Arg-Glu-Arg (SPIRER), or Ser-Pro-He-Arg-Lys (SPIRK), or Ser-Pro-fle-Lys-Lys (SPIKK), or Ser-Pro-Ile-Arg-Arg-Pro (SPIRRP), or Ser-Pro-Pro-Arg-Arg (SPPRR), or Ser-Pro-Iso-Pro-Arg (SPIPR), or Ser-Pro-Arg-Pro-Arg (SPRPR), or Ser-Pro-Ile-Arg (SPIR), or Ser-Pro-ne-Arg-Arg (SPIRR), or Ser-Cys-ile-Arg-Arg, (SCIRR), or Ser-Pro-Ile-Arg-Pro-Arg-Pro (SPIRPRP), or Ser-Cys-Ile-Arg-Pro-Arg-Pro (SCPIRPRP), or Ser-Pro-Arg-Arg-Pro-Arg-Thr (SPRRPRT), or Ser-Pro-Phe-Arg-Pro-Lys-Leu (SPFRPKL), or Ser-Pro-Pro-Arg-Arg-Pro (SPPRRP), or Ser-Pro-πe-Arg-Arg-Glu (SPIRRE), or Ser-Pro-Pro-Arg-Pro-Pro (SPPRPP), or Ser-Pro-Pro-Arg-Pro-Arg (SPPRPR), or Ser-Pro-Pro-Trp-Tip-Pro (SPPWWP), or Ser-Pro-Pro-Trp-Arg-Pro (SPPWRP), or Ser-Pro-Pro-Arg-Trp-Pro (SPPRWP), or Ser-Pro-Pro-Arg-Trp-Pro (SPPRWP), or Ser-His-Trp-Arg-Arg-Tφ (SHWRRW), or Ser-His-Tφ-Arg-Lys (SHWRK), or Ser-His-Tφ-Arg-Arg (SHWRR), or Thr-Ala-Ile-Arg-Pro-Arg-Lys (TAIRPRK), Ser-Thr-Arg-Arg-Pro-Arg-Pro (STRRPRP), Gly-Pro-He-Arg-Pro-Arg-Pro (GPIRPRP), or Leu-Pro-Phe-Arg-Gln-Arg-Pro (LPFRQRP), or HEET Ser-Arg-Ser-Arg-His-Asn-Ala (SRSRHNA), or Ile-Pro-Ile-Arg-Pro-Arg-Arg (IPIRPRR), or Ser-Thr-Arg-Arg-Pro-Arg-Pro (STRRPRP), or Thr-Ala-He-Arg-Pro-Arg-Lys (TAIRPRK), or Trp-Arg-Tφ-Arg-Trp-Arg (WRWRWR), or Gln-Pro-Ile-Arg-Arg (QPIRR), or Ser-His-Tip-Gln-Gln (SHWQQ), or Ser-Ala-Leu-Arg-Pro-Arg-Lys (SALRPRK). Also contemplated according to the invention is additions comprising more than 7 amino acids, such as from 8 to 15 amino acids. Such peptides can be generaUsed as: a-b-c-d-e-f-g-h (8 amino acid peptide) a-b-c-d-e-f-g-h-i (9 amino acid peptide) a-b-c-d-e-f-g-h-i-j (10 amino acid peptide) a-b-c-d-e-f-g-h-i-j-k (11 amino acid peptide) a-b-c-d-e-f-g-h-i-j-k-1 (12 amino acid peptide) a-b-c-d-e-f-g-h-i-j-k-l-m-(13 amino acid peptide) a-b-c-d-e-f-g-h-i-j-k-l-m-n(14 amino acid peptide) a-b-c-d-e-f-g-h-i-j-k-1-m-n-o (15 amino acid peptide). a to o may be any of the twenty amino acids mentioned above. The a-g stretch may be as defined above in relation to a peptide addition comprising 1 to 7 amino acid residues. h, i, j, k, 1, m, n, o may as mentioned above be any amino acid, preferably any of the foUowing amino acids: Arg, Lys, Ala, Val, Tφ, De, Phe, Ser or Pro. Specific examples of such additions are Usted below: Arg-Pro-Arg-Pro-Arg-Pro-Arg-Pro (RPRPRPRP), or Ser-Ser-Thr-Arg-Arg-Ala-Ser-Pro-πe-Lys-Lys (SSTRRASPIKK), or Ala-Tφ-Tφ-Pro-Ser-Pro-Ile-Arg-Pro-Arg-Pro (AWWPSPIRPRP), or Ala-Pro-Pro-Pro-Arg-Pro-Arg-Pro-Arg-Pro-Arg-Pro (APPPRPRPRPRP), or Ala-Pro-Pro-Pro-Arg-Thr-Arg-Pro-Arg-Pro-Arg-Ser(APPPRTRPRPRS), or Ser-Pro-Lys-Arg-Lys-Pro-Arg-Pro (SPKRKPRP), or Ser-Gln-Arg-πe-Lys-Gln-Arg-Ile-Lys (SQRIKQRIK), or Ser-Pro-Pro-Pro-Arg-Pro-Arg-Pro (SPPPRPRP), or Ser-Pro-De-Arg-Pro-Arg-Pro-Arg-Pro-Arg SPD PRPRPR, or Ser-Pro-πe-Arg-Lys-Ala-Trp-Trp-Pro (SPIRKAWWP), or Ala-Pro-Pro-Pro-Lys-Ala-Ser-Pro-Arg-Gln-Arg-Pro (APPPKASPRQRP), or Ser-Pro-Ile-Arg-Pro-Arg-Pro-Ser-Pro-Ile-Arg-Pro-Arg-Pro-Arg (SPIRPRPSPIRPRP), or Ser-Pro-Pro-Arg-Trp-Pro-Arg-Arg (SPPRWPRR), or Ser-Pro-Pro-Arg-Trp-Pro-Arg-Tφ (SPPRWPRW), or Ser-Pro-Pro-Arg-Trp-Pro-Trp-Arg (SPPRWPWR), or Ser-Pro-Pro-Trp-Arg-Pro-Arg-Arg (SPPWRPRR), or Ser-Pro-Pro-Trp-Trp-Pro-Arg-Tφ (SPPWWPRW, or Ser-Pro-Pro-Tφ-Tφ-Pro-Tφ-Arg (SPPWWPWR), or Ser-Pro-Pro-Tφ-Tφ-Pro-Tφ-Tφ (SPPWWPWW), or Ser-Pro-Pro-Trp-Pro-Arg-pro-Arg-Pro (SPPWPRPRP), or Ala-Pro-Pro-Pro-Arg-Pro-Arg-Leu-Leu-Pro-Ile-Ser (APPPRPRLLPIS), or Ala-Pro-Pro-Pro-Thr-Arg-Gln-Arg-Gln-Ser-Pro (APPPTRQRQSP), or Ala-Pro-Pro-Pro-Arg-Thr-Ile-Pro-Arg-Ser-Ser-Pro (APPPRTIPRSSP). In any of the above specified peptide additions (whether comprising 1 to 7 or 1 to 15 amino acid residues) in which the position "a" is a Ser, Ala, Arg, Lys or Pro, the Ser may be replaced with an Ala, Arg, Lys or Pro, the Ala with a Ser, Arg, Lys or Pro and the Arg, Lys or Pro with a Ala or Ser. It is to be emphasised that the above peptide addition may be at either the N-terminal and/or the C-terminal. Examples of modified Upolytic enzymes with both a N- and a C- terminal peptide addition include aU combinations of the peptide additions SpecificaUy men¬ tioned above. Two specific examples of such are the N-terminal addition SPIRPRP together with the C-terminal addition RRP or RR. In addition to the above specified peptide additions it has been found that a suitable pep¬ tide addition may simply be constituted by or comprise a part of or the entire propeptide se¬ quence normaUy associated with the parent Upolytic enzyme in question. Thus, for instance in relation to first wash H. lanuginosa Upolytic enzyme variants a suitable peptide addition may comprise or be constituted of SPIRR - i.e. part of the normal propeptide sequence of the H. lanuginosa Upolytic enzyme sequence. T If the peptide addition is inserted into the non-stmctural part of the parent enzyme, it may replace one or more of the amino acid residues of said non-stmctural part. For instance, the peptide addition may replace one or more amino acid residues occupying the first, e.g. 1-5, amino acid residues of the N-terminal end and/or the last, e.g. 1-5, amino acids of the en- zyme (i.e. the 1-5 amino acid residues of the C-terminal end). For instance, the peptide addi¬ tion may replace amino acid residue(s) 1 and/or 2 and/or 3 and/or 4, and/or 5, etc. from ei¬ ther end of the parent enzyme. When the parent enzyme is a variant of the H. lanuginosa Upase which comprises amino acid modifications in its stmctural, mature part it has been of particular interest to combine any of the above peptide additions (appUed in the N-terminal) with a deletion of the first amino acid residue of the mature parent enzyme, i.e. the IE. Methods of applying a peptide addition to a parent lipolytic enzyme Although a first wash Upolytic enzyme of the invention may be obtained by adding (fusing or inserting) a syntheticaUy produced peptide addition into the parent Upolytic enzyme in question, it is presently prefened that the modified enzyme of the invention is prepared by i) modifying the nucleotide, preferably DNA, sequence encoding the parent enzyme so as to encode the desired peptide addition appUed to the N- and/or the C-terminal end(s) of the par¬ ent enzyme (e.g. by inserting a nucleic acid (preferably DNA) sequence encoding the peptide addition at the relevant location in the nucleic acid (preferably DNA) sequence encoding the parent enzyme), u) expressing the resulting modified nucleic acid (preferably DNA) sequence in a suitable expression system, and in) recovering the resulting modified enzyme. In the present context, the term "appUed to" is intended to indicate that the addition is fused to the N- and/or C-terminal end (e.g. to the first or last amino acid residue) of the ma- ture enzyme or inserted into a non-stmctural part of the N-terminal and/or C-terminal end of the mature enzyme. Many enzymes are expressed as "prepro-enzymes", i.e. as enzymes consisting of the mature enzyme, a secretory signal peptide (i.e. prepeptide) and a pro-peptide. The prepro- enzyme is processed intraceUularly to be secreted into the fermentation medium, from which the mature enzyme can be isolated and/or purified. The peptide addition to the parent enzyme can be carried out by applying nucleic acid sequences encoding the desirable peptide additions upstream (for N-terminal peptide additions) and/or downstream (for C-terminal peptide addi¬ tions) to the DNA sequence encoding the parent enzyme. The insertion should be performed in such a way that the desired modified enzyme (i.e. having the desired peptide addition(s)) is expressed and secreted by the host ceU after tran- scription, translation, and processing of the enzyme. The term "processing" means in this context removal of pre- and pro-peptides (except, of course, when the pro-peptide is identical to the desired peptide addition. This wiU be dealt with further below). Downstream sequences (encoding a C-terminal addition) can be inserted between the DNA sequence encoding the parent enzyme and the terminating codon. However, if the un- processed DNA sequence comprises a pro-peptide encoding DNA sequence at the C-terminal end the insertion addition of the DNA sequence encoding the peptide addition can also take place between the DNA sequences encoding the pro-peptide and the mature enzyme, respec¬ tively. In most cases it is possible to extend the parent enzyme upstream by inserting a DNA se- quence encoding the peptide addition between the DNA sequence encoding the pro-peptide or the prepeptide (if no prosequence is present) and the DNA sequence encoding the mature en¬ zyme. The DNA sequence encoding the peptide addtion in question shaU, of course, be chosen so as to match the codon preferences of the expression system intended for the production of the first wash Upolytic enzyme of the invention. The insertion addition of a DNA sequence encoding the peptide addition can be canied out by any standard techniques known by any skilled person in the field of molecular biology, cf., e.g. Sambrook et al., 1989). This include, e.g., the polymerase chain reaction (PCR) us¬ ing specific primers, for instance described in US patent 4,683,202 or R.K. Saiki et al., (1988), Science, 239, 487-491. How to provide for the expression and secretion of adjacent DNA sequence(s) will be described below. In connection with the present invention it has been found that some host ceUs may be less suited for the production of a first wash Upolytic enzyme comprising a peptide addtion, in that part or aU of the peptide addition(s) may be cut off during the posttranslational or other proc- essesing performed by the host ceU. Accordingly, the term "suitable expression system" is intended to indicate an expression system (host ceU and optionaUy expression vector) which aUows for at least a portion of the intact desired first wash Upolytic enzyme comprising the peptide addition to be produced, i.e. an expression system which does not, e.g. as part of the posttranslational or other processing by the host ceU of choice, remove part or aU of the pep¬ tide addition (and thereby produce the enzyme without the desired peptide addition). Ex¬ pressed differently, the expression system (including the host ceU, cultivation conditions and/or recovery conditions) are preferably selected so that at the most a partial processing of the pre, pro or prepro-form of the Upolytic enzyme occur resulting in that at least 5 % , such as at least 10%, such as at least 15%, such as at least 20%, such as at least 25%, such as at least 50% , such as at least 75 % of the produced enzyme molecules comprise the desired peptide addition, e.g. the entire pro-sequence or a substantial part thereof. TypicaUy, the expression system to be used is devoid of or reduced in one or more proteolytic activities exerting the undesired posttranslational processing. The choice of expression system and thus host ceU will depend on the Upolytic enzyme to be produced as wiU be discussed in detaU further below. While care must be exerted to select a proper expression system for producing a first wash Upolytic enzyme of the invention which comprises a peptide addition at its N- and/or C- terminus (in particular when a modified DNA sequence is used for the production), it has been found that when the peptide addition is the propeptide or part thereof normaUy associated with the parent Upolytic enzyme in question, the peptide addition (thus constituting a part of or the entire propeptide sequence) may be appUed by expressing a DNA sequence encoding the parent Upolytic enzyme in question in an expression system which is incapable of proc- essing the translated polypeptide in the normal manner, and thereby results in the production of an enzyme which comprises a part of or the entire propeptide or a siπύlar peptide sequence associated with the mature protein prior to its processing. In this case, the propeptide or simi¬ lar peptide sequence constitutes the peptide addition. The pro-peptide or simUar peptide se¬ quence may be heterologous or homologous to the parent enzyme and can be present in both the N- and C-terminal of the parent enzyme. Accordingly, if a suitable stretch of amino acids is already encoded in the prepro form of the parent enzyme and this stretch of amino acids is cut off in the processing of the enzyme by a given expression system, the peptide addition can be appUed by changing the expression host system to a system in which said processing of said stretch of amino acids does not occur or modify the gene sequence to eUminate the post-translation processing, e.g. by saturating the processing enzyme(s) with one or more copies of a pro-like peptide (such as one of the pep¬ tide additions shown herein) or by changing the pro-peptide sequence, e.g. to remove a post- translational processing site. In such a case the secretory signal pre-peptide will be cut off during or after the secretion, resulting in a modified enzyme consisting of the parent enzyme comprising the pro-peptide or part thereof or a similar peptide sequence encoded by the corre¬ sponding DNA sequence, i.e. a Upolytic enzyme being extended at either its N-terminal or C- terminal end. In other words, the first wash Upolytic enzyme of the invention which comprises a peptide addtion may be designed and/or produced by a method comprising: cultivating a host ceU transformed with a DNA sequence encoding the parent Upolytic en¬ zyme including its (pre)pro sequence under conditions suitable for production of the enzyme comprising at least a part of the entire pro(pre)-sequence, the host ceU being one which is in¬ capable or inefficient in the processing of the pro-enzyme to be expressed into the mature en¬ zyme, and recovering and optionaUy purifying the resulting modified enzyme. The DNA sequence encoding the parent Upolytic enzyme may be present on an expression vector, when transformed into the host ceU. The host ceU may be of a different origin than the parent enzyme, e.g. of another genus than the one from which the parent enzyme is derived, or may have another posttranslational processing machinery than the source of the parent enzyme. Yeast ceUs have been found of particular use for applying peptide additions (in the form of the propeptide or a part thereof) to parent filamentous fungal Upolytic enzymes, in particular to H. lanuginosa Upolytic enzyme variants, due to the different processesing system of the yeast ceUs as compared to the fila¬ mentous fungal ceUs. Examples of suitable yeast ceUs for said purpose are ceUs derived from a strain of Saccharomyces sp., in particular Saccharomyces cerevisiae, or a strain of Han¬ senula sp.. In an altemative and highly prefened embodiment the first wash Upolytic enzyme of the invention comprising a peptide addition has been designed and/or produced by a method com¬ prising the foUowing steps: a) subjecting a DNA sequence encoding the parent Upolytic enzyme with a peptide addition to localized random mutagenesis in the peptide addition or in a non-stmctural part of the C- terminal or N-terminal end of the parent enzyme, b) expressing the mutated DNA sequence obtained in step a) in a host ceU, c) screening for host ceUs expressing a mutated Upolytic enzyme which has an improved per¬ formance as compared to the parent Upolytic enzyme, d) selecting a mutated Upolytic enzyme among those resulting from step c) which, when pres¬ ent in detergent composition A and or B with 12500 LU/1 detergent, is capable of remov¬ ing at least 15% more lard from a lard stained swatch, than the same detergent composition without the enzyme, in a one cycle wash assay as disclosed herein. By this approach a number of highly advantageous first wash Upolytic enzymes with different peptide additions have been created. The peptide addition present on the DNA sequence to be mutagenized may be constituted by or comprise the prosequence or a part thereof normaUy associated with the parent Upolytic enzyme or may be any other peptide addition, e.g. one of the peptide additions exempUfied above. Each of steps a)-d) may be carried out as described in the sections further below entitied "Random mutagenesis" and "Localized Random Mutagenesis". The parent lipolytic enzyme The parent Upolytic enzyme to be modified in accordance with the invention may be of any origin. Thus, the enzyme may be of mammaUan, plant, vertebrate or any other origin. However, it is presently prefened that the enzyme is of microbial origin in that a number of microbial strains have been found to produce enzymes of particular use for detergent purposes. More SpecificaUy, the parent Upolytic enzyme may be derived from a fungus, i.e. a yeast or a filamentous fungus. For instance, the enzyme may be derived from a filamentous fungus of the class of Plectomycetes, preferably the order of Eurotiales and more prefera¬ bly the family like Eremascaceae, Monoascaceae, Pseudoeurotiaceae and Trichoco- maceae, the latter containing genera like Emericella, Aspergillus, Penicillium, Eupenicil- lium, Paecilomyces, Talaromyces, Thermoascus and Sclerocleista. More specifically, the parent enzyme may be one which is derivable from a strain of a Humicola sp. , e.g. H. brevis- pora, H. lanuginosa, H. brevis var. thermoidea and H. insolens (US 4,810,414), a strain of a Rhizomucor sp., e.g. Rh. miehei (EP 238023), a strain of a Rhizopus sp., e.g. R. delemar (Hass et al., (1991), Gene 109,107-113), R. niveus (Kugimiya et al., (1992) Biosci.Biotech. Biochem 56, 716-719) or R. oryzae, a strain of a Candida sp., e.g. C. cylindracea (also caUed C. rugosa) or C. antarctica (WO 88/02775) or C. antarctica Upase A or B (EP 214 761), a strain of n Fusarium sp., e.g. F. oxysporum (EP 130,064) or F. solanipisi (WO 90/09446) or variants thereof (WO 94/14964), F. solanipisi (GB 2 296011) a strain of a Venturia spp., e.g. V. inaequalis, a strain of a Colletotrichum spp., e.g. C. gloeosporioides, or C. lagenarium, a strain of Geotricum, e.g. G. candidum (Schimada et al., (1989), J.Biochem., 106, 383-388), a strain of Aspergillus, e.g. A. niger, or an Aspergillus sp. Upolytic enzyme variant (EP 167,309), or a strain of a Penicillium spp., e.g. P. spinulosum or P. camembertii (Yamaguchi et al., (1991), Gene 103, 61-67). In the present context, "derivable from" is intended not only to indicate an enzyme produced by a strain of the organism in question, but also an enzyme encoded by a DNA sequence isolated from such strain and produced in a host organism transformed with said DNA sequence. Furthermore, the term is intended to indicate an enzyme which is encoded by a DNA sequence of synthetic and/or cDNA origin and which has the identifying characteris¬ tics of the enzyme in question. FinaUy, the term is intended to embrace variants of the enzyme, e.g. carrying one or more mutations as compared to the naturaUy occurring enzyme, or homologous enzymes which may be naturaUy-occuπing enzymes produced by other strains or organisms, which, e.g. may be isolated by hybridization to oUgonucleotide probes prepared on the basis of the amino acid or DNA sequence of any of the above enzymes (the hybridization conditions involving presoaking in 5xSSC and prehybridizing for 1 h at ~40°C in a solution of 20% formamide, 5xDenhardt's solution, 50 mM sodium phosphate, pH 6.8, and 50 g of denatured sonicated calf thymus DNA, foUowed by hybridization in the same solution supplemented with 100 M ATP for 18h at ~40°C, or other methods described by Sambrook et al., 1989) or which is immunologicaUy cross-reactive with said enzymes (e.g. as determined by the method of Hudson et al., 1989). Of particular interest as a parent Upolytic enzyme is one derivable from a strain of H. lanuginosa, e.g. the H. lanuginosa strain DSM 4109, e.g. the mature form of the enzyme described in EP 305 216 or a variant thereof as described in WO 92/05249, WO 94/01541 , WO 94/14951, WO 94/25577, PCT/DK94/00079 (aU from Novo Nordisk A/S), which are hereby incorporated by reference. Throughout the present application the name Humicola lanuginosa has been used to identify one preferred parent enzyme, i.e. the one mentioned immediately above. How¬ ever, in recent years H. lanuginosa has also been termed Thermomyces lanuginosus (a species introduced the first time by Tsiklinsky in 1989) since the fungus show moφho¬ logical and physiological similarity to Thermomyces lanuginosus. Accordingly, it will be understood that whenever reference is made to H. lanuginosa this term could be replaced by Thermomyces lanuginosus. The DNA encoding part of the 18S ribosomal gene from Thermomyces lanuginosus (or H. lanuginosa) have been sequenced. The resulting 18S sequence was compared to other 18S sequences in the GenBank database and a phyloge¬ netic analysis using parsimony (PAUP, Version3.1.1, Smithsonian Institution, 1993) have also been made. This clearly assigns Thermomyces lanuginosus to the class of Plec¬ tomycetes, probably to the order of Eurotiales. According to the Entrez Browser at the NCBI (National Center for Biotechnology Information), this relates Thermomyces lanugi¬ nosus to families like Eremascaceae, Monoascaceae, Pseudoeurotiaceae and Trichoco- maceae, the latter containing genera Hke Emericella, Aspergillus, Penicillium, Eupenicil- Hum, Paecilomyces, Talaromyces, Thermoascus and Sclerocleista. The parent Upolytic enzyme to be modified in accordance with the present in¬ vention may be derivable from a bacterium. For instance, the DNA sequence encoding the parent Upolytic enzyme may be derivable from a strain of Pseudomonas spp., such as Ps. ce¬ pacia, Ps. alcaligenes, Ps. pseudoalcaligens, Ps. mendocina (also termed Ps. putida), Ps. syringae, Ps. aeroginosa, Ps. wisconsinensis (WO 96/12012) or Ps.fragi, a strain of Bacillus spp., e.g. B. subtilis or B. pumilus or a strain of Streptomyces sp., e.g. S. scabies. In connection with the Pseudomonas sp. Upases it has been found that Upases from the foUowing organisms have a high degree of homology, such as at least 60% homology, at least 80% homology or at least 90% homology, and thus are contemplated to belong to the same family of Upases: Ps. ATCC21808, Pseudomonas sp. Upase commerciaUy avaUable as Liposam ® , Ps. aeruginosa EF2, Ps. aeruginosa PAC1R, Ps. aeruginosa PAO1, Ps. aerugi¬ nosa TE3285, Ps. sp. 109, Ps. pseudoalcaligenes Ml, Ps. glumae, Ps. cepacia DSM3959, Ps. cepacia M-12-33, Ps. sp. KWI-56, Ps. putida IFO3458, Ps. putida IFO12049 (GUbert, E. J., (1993), Pseudomonas Upases: Biochemical properties and molecular cloning. Enzyme Microb. Technol., 15, 634-645). The species Pseudomonas cepacia has recently been reclas- sified as Burkholderia cepacia, but is termed Ps. cepacia in the present apphcation. Specific examples hereof include a Pseudomonas Upolytic enzyme, e.g. Ps. fragi, Ps. stutzeri, Ps. cepacia and Ps. fluorescens (WO 89/04361), or Ps. plantarii or Ps. gladioli (US 4,950,417) or Ps. alcaligenes and Ps. pseudoalcaligenes (EP 218 272, EP 331 376, or WO 94/25578 (disclosing variants of the Ps. pseudoalcaligenes Upolytic enzyme with the mutation M21S, M21L or M21A), the Pseudomonas sp. variants disclosed in EP 407 225, or a Pseudomonas sp. Upolytic enzyme, such as the Ps. mendocina Upolytic enzyme described in WO 88/09367 and US 5,389,536 or variants thereof as described in US 5,352,594. Other specific examples include a Bacillus Upolytic enzyme, such as the Upolytic enzyme from B. subtilis (Dartois et al., (1993) Biochemica et Biophysica acta 1131, 253-260) or B. stearothermophilus (JP 64/7744992) or B. pumilus (WO 91/16422) and a Chromobacterium Upolytic enzyme (especiaUy one derivable from C. viscosum). Specific examples of readUy avaUable commercial Upolytic enzyme which may serve as parent Upolytic enzymes according to the invention include Lipolase®, Lipolase® Ultra (avaUable from Novo Nordisk A S). Examples of other Upolytic enzymes SpecificaUy contemplated to be modifiable according to the invention are Lumafast®, i.e. a Ps. mendocina Upolytic enzyme and Lipomax®, i.e. a Ps. alcaligenes Upolytic enzyme, a Fusarium solani Upase (cutinase) from UnUever, a Bacil¬ lus sp. Upase from Solvay enzymes (US 5427936, EP 528828); and Liposam ® , (a Ps. men¬ docina Upase from Showa Denko) and further the Pseudomonas sp. Upase described in WO 95/06720 which have been sequenced and found to have the amino acid sequence shown in SEQ ID NO 99. It is to be emphasized that the parent Upolytic enzyme to be modified according to the invention may be any of the above mentioned Upolytic enzymes and any variant, modification, or truncation thereof. Examples of such parent enzymes which are SpecificaUy contemplated include the enzymes described in WO92/05249, WO 94/01541, WO 94/14951, WO 94/25577, WO 95/22615 and a protein engineered Upase variants as described in EP 407 225; a protein engineered Ps. mendocina Upase as described in US 5,352,594; a cutinase variant as described in WO 94/14964; a variant of an Aspergillus Upolytic enzyme as described in EP patent 167,309; and Pseudomonas sp. Upase described in WO 95/06720. Specific first wash H. lanuginosa Upolytic enzyme variants For ease of reference specific variants of the invention are described by use of the fol¬ lowing nomenclature: Original amino acid(s):position(s): substituted amino acid(s) According to this nomenclature, for instance the replacement of aspartic acid by valine in position 96 is shown as: Asp 96 Val or D96V a deletion of aspartic acid in the same position is shown as: T Asp 96 * or D96* and insertion of an additional amino acid residue such as lysine is shown as: Asp 96 ValLys or D96VK Multiple mutations are separated by pluses, i.e.: Asp 96 Val + Glu 87 Lys or D96V+E87K representing mutations in positions 96 and 87 replacing aspartic acid and glutamic acid by valine and lysine, respectively. When one or more altemative amino acid residues may be inserted in a given position it is indicated as D96V,N or D96V or D96N Furthermore, when a position suitable for modification is identified herein without any specific modification being suggested, it is to be understood that any amino acid residue may be substituted for the amino acid residue present in the position. Thus, for instance, when a modification of an aspartic acid in position 96 is mentioned, but not specified, it is to be understood that the aspartic acid may be deleted or replaced by any other amino acid, i.e. any one of R,N,A,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V, or a further amino acid residue inserted at that position. FinaUy, when a mutation of the parent H. lanuginosa Upolytic enzyme is identified herein, it is intended to be understood to include a mutation of an amino acid residue occupying a homologous position in a Upolytic enzyme which has substantiaUy the same stmcture as or a stmcture or amino acid sequence which can be aUgned with that of the H. lanuginosa enzyme (e.g. Rhizopus oryzae, Rhizomucor miehei, Absidia sp. and Penicillium camembertii Upolytic enzymes mentioned herein). The homologous position can easUy be identified by comparison between the stmctures The H. lanuginosa Upolytic enzyme variants may be characterized by being a combination of at least two different parent variants which indviduaUy have been found or indicated to have a good wash performance or otherwise interesting properties as described above. The good wash performance may e.g. be determined as described in Example 6. The combination between the parent variants may be random or specific.In connection with the present invention it has been found that particularly interesting results are obtained when the parent variants to be combined contains a mutation in at least one, but preferably more of the Mowing positions: 1, 2, 3, 4, 5, 19, 49, 53, 56, 57, 59, 62, 83, 85, 90, 94, 96, 97, 99, 101, 102, 111, 116, 126, 127, 137, 167, 170, 181, 187, 210, 221, 225, 234, 239, 249, 252 256, 263, 264, 267, such as at least one or preferably more of the foUowing mutations: ElK, EIS, V2G, S3T, Q4P, D5E, A19T, A49P ,Y53C, E56K, D57G, G59V, D62R, S83T, S85F, I90F, N94K, F95L, D96A, D96H, D96L, L97M, E99K, N101S, D102Y, Dil IN, S116P, Q126R, K127C, D137G, D167G, S170P, F181L, V187A, E210K, E210V, W221L, W221A, G225P, D234R, D234Y, E239C, Q249R, I252L, P256T, G263A, L264Q, T267R. It wiU be understood that the above mutations or mutated positions may be present on the same parent Upolytic enzyme, but preferably on various of the different parent Upolytic enzymes to be combined. In particular, it has been found that a first wash H. lanuginosa Upolytic enzyme of the invention may be a combmation of at least two of the foUowing parent H. lanuginosa Upolytic enzyme variants or parts of these variants: (a) E56R+D57L+I90F+D96L+E99K (b) E56R+D57L+V60M+D62N+S83T+D96P+D102E (c) D57G+N94K+D96L+L97M (d) E87K+G91A+D96R+I100V-l-E129K+K237M+I252L+P256T-l-G263A+L264Q (e) E56R+D57G+S58F+D62C+T64R+E87G+G91A+F95L+D96P+K98I (f) E210K (g) S83T+N94K+D96N (h) E87K+D96V (i) N94K+D96A (j) E87K+G91A+D96A (k) D167G+E210V (1) S83T+G91A+Q249R (m) E87K+G91A (n) S83T+E87K+G91A+N94K+D96N+D11 IN (o) N73D+E87K+G91A+N94I+D96G (p) L67P+I76V+S83T+E87N+I90N-l-G91A-l-D96A+K98R (q) S83T+E87K+G91A+N92Η+N94K+D96M (s) S85P+E87K+G91A+D96L+L97V (t) E87K+I90N+G91A+N94S+D96N+I100T (u) D4V+S54P+F80L+S85T+D96G+R108W+G109V+DlllG+S116P+L124S+ V132M+V140Q+V141A+F142S+H145R+N162T+I166V+F181P+F183S+R205G+A 243T+D254G+F262L, (v) N94K, D96A, Q249R, 5 (w) E87K, G91A, D96W, D102N Methods suitable for combining different parent variants are described below in the section in entitied "Combination of DNA sequences encoding Upolytic enzymes". A particular suitable method is the one described in Materials and Methods section herein. In another embodiment the first wash Upolytic enzyme of the invention is a variant of the H. 10 lanuginosa Upolytic enzyme (the amino acid sequence of which is shown in SEQ ID No.1) which comprises a mutation in at least one, but preferably more of the foUowing positions: 1, 2, 3, 4, 5, 19, 49, 53, 56, 57, 59, 62, 83, 85, 90, 94, 96, 97, 99, 101, 102, 111, 116, 126, 127, 137, 167, 170, 181, 187, 210, 221, 225, 234, 239, 249, 252 256, 263, 264, 267, such as at least one or preferably more of the foUowing mutations, ElK, EIS, V2G, S3T, Q4P, 15 D5E, A19T, A49P, Y53C, E56K, D57G, G59V, D62R, S83T, S85F, I90F, N94K, F95L, D96A, D96H, D96L, L97M, E99K, N101S, D102Y, Dil IN, S116P, Q126R, K127C, D137G, D167G, S170P, F181L, V187A, E210K, E210V, W221L, W221A, G225P, D234R, D234Y, E239C, Q249R, I252L, P256T, G263A, L264Q, T267R In a more specific embodiment the first wash Upolytic enzyme of the invention is a variant of 20 the H. lanuginosa Upolytic enzyme (the amino acid sequence of which is shown in SEQ ID No.1), in which at least one of the following amino acid residues has been replaced with another amino acid residue: A49, G59, S85, 190, S116, Q126, D137, S170 or W221. Although the above identified amino acid residues may be replaced by any other of the 25 19 possible amino acid residues it is preferred that the amino acid residue is replaced as foUows: A49P, G59V, S85F, I90F, S116P, Q126R, D137G, S170P or W221L or by an amino acid residue belonging to the same charge group (cf the definition below) as that of the inserted amino acid residue, e.g. A49T instead of A49P. If a negatively charged amino acid residue is 30 replaced, e.g. D137, it is preferred that it is replaced by an amino acid residue belonging to the positive charge group or the neutral group, e.g. D137G,N,K, as defined below: Negative charge group: D,E Positive charge group: K,R,H Neutral group: I,C,S,T,P,W,M,G,A,P,N,Y,Q,L,V It is contemplated that a variant comprising a mutation in the foUowing positions is capable of exhibiting first wash activity or improved wash performance: D57X+N94(K or R)+D96X+L97X+Q249(K or R) N94(K or R)+D96X+L97X+Q249(K or R) N94(K or R)+D96X+Q249(K or R) D137X+D167X+E210X+W221X D137X+D167X+E210X I90X+D96X+E99X+V187X I90X+D96X+E99X I90(F or W or Y)+D96X+E99X E56X+D57X+D62X+S85X+D96X+D102X+E210X N94(K or R)+F95L+D96X+D234X, in which X, may be any amino acid residue and may be identical, pairwise identical or different. A H. lanuginosa Upolytic enzyme variant of the invention may comprise the foUowing mutations: D57G+N94K+D96L+Q249R D57G+N94K+D96L+S116P+Q249R D57G+G59V+N94K+D96L+Q249R D57G+N94K+D96L+S 116P+S 170P+Q249R D57G+G59V+N94K+D96L+S170P+Q249R D57G+N94K+D96L+S170P+Q249R D167G+E210V+Q249R E56K+D167G+E210V D137G+D167G+E210V+Q249R D167G+E210V+W221L+Q249R D57G+N94K+F95L+D96H,L+Q249R D57G+N94K+D96L+E210K D57G+G59V+N94K+D96L+S116P+S170P+Q249R S3R+D137G+D167G+E210V+W221L D137G+D167G+E210V+W221L+N233R S3R+I90F+D96L+E99K+V187A+Q249R I90F+D96L+E99K+V187A+D233R I90F+D96L+E99K+V187A+D234Y I90F+D96L+E99K+V187A+T231R I90F+D96L+E99K+V187A D62R+I90F+D96L+E99K+V187A I90F+D96L+E99K+V187A+N200R+R209A I90F+D96L+E99K+V187A+T199R+N200R+R209A D57G+D62R+N94K+D96L+Q249R D57G+N94K+D96L+N200R+R209A+Q249R D57G+N94K+D96L+T199R+N200R+Q249R I90F+D96L+E99K+V187A+T199R D57G+N94K+D96L+T199R+R209A+Q249R I90F+D96L+E99K+V187A+Q249R I90F+D96L+E99K+V187A+P253R I90F+D96L+E99K+D137G+D167G+V187A+Q249R I90F+D96L+E99K+D137G+V187A+Q249R D96L+E99K+V187A+Q249R V2P+N94K+D96L+Q249R V2W+S3R+N94K+D96L+Q249R V2R+S3R+N94K+D96L+Q249R V2R+S3R+N94K+D96L+Q249R V2R+S3W+N94K+D96L+Q249R V2W+S3R+N94K+D96L+Q249R N94K+D96L+Q249R V2G+S3T+D57G+N94K+D96L+L97M+Q249R V2G+S3T+Q4P+D5E+D57G+N94K+D96L+L97M+Q249R V2G+D5Q+L6M +D57G+N94K+D96L+L97M+Q249R The foUowing variants are of particular interest: D57G+G59V+N94K+D96L+L97M+S116P+S170P+Q249R A49P+D167G+E210V E56K+D57G+D62R+S83T+S85F+D96L+D102Y+E210K D57G+N94K+D96L+L97M+Q249R D137G+D167G+E210V+W221L N94K+F95L+D96H+N101S+F181L+D234Y+E52L+P256T+G263A+L264Q I90F+D96L+E99K+V187A N94K+D96A+Q249R A19P+D167G+E210V+W221L N94K+D96L+L97M+Q249R D57G+N94K+D96L+Q249R I90F+D96L+E99K+D137G+V187A N94K+D96L+E99K+Q249R N94K+D96L+E99K+T231R+N233R+D234R+Q249R N94K+D96L+E99K+D111N+F211A+G225P+Q249R+T267R N94K+D96L+E99K+D111N+F211A+G225P+T231R+N233R+D234R+Q249R+ T267R E1K+N94K+D96L+E99K+Q249R N94K+D96L+K223R+Q249R N94K+D96L+E99K+N233R N94K+D96L+E99K+T231R+N233R+Q249R N94K+D96L+E99K+N233R+Q249R N94K+D96L+E99K+D234R+Q249R The variant of the invention may advantageously comprise an additional mutation in position El , the mutation being a deletion of El or a replacement of E by any other amino acid residue, in particular P or S. In addition the above specific variants may comprise any of the N-terminal or C- terminal peptide extensions discussed herein, specific examples of which are SPIRR, TAIRPRK, SPIRPRP, SPPRRP, RP, GPIRPRP, SRSRHNA, SALRPRK, STRRPRP, SPRRPRT, APPPRPRPLLPIS, SPIRK, SPPRPRP, WP, SPPPRPRP, SPIRRP, APPPRPRPRPR or SPIRPR. An N-terminal extension is e.g. appUed to the amino acid residue El of the mature parent Upase or is appUed to amino acid residue 2-20, such as 2, 3, 4 or 5 of the mature parent enzyme, the residue El (and optionaUy more amino acid residues of the non-stmctural part of the parent enzyme, e.g. amino acid residues within the 2-20 N- terminal part of the mature parent enzyme) being deleted. In addition, the peptide addition may be appUed so that the one or more of the last amino acid residues of the peptide extensions mentioned herein replaces the amino acid residue(s) of the mature parent enzyme occupying position 1, and optionaUy 2 and further positions. For instance, the peptide extension "SPPRRP" may be appUed by substituting El of the mature parent H. lanuginosa Upase with the last "P" of the peptide addition and substituting the wUdtype propeptide "SPIRR" with "SPPRR". When no replacements are to be performed in the N-terminal part of the mature parent enzyme, the N-terminal addition may be appUed either as a result of the variants having been expressed in S. cerevisiae (if the N-terminal extension is identical to (a part of) the propeptide of the parent enzyme, or more preferably by the relevant modification of the part of the DNA sequence encoding the parent enzyme, which encodes the (pre)pro sequence or another sequence downstream of the codon encoding amino acid residue 1 of the mature parent enzyme. The presently most preferred variants of the inventions include: SPIRPRP-I-D57G+N94K-I-D96L+Q249R SPPRRP+I90F+D96L+E99K+D137G+V187A SPERPRP+N94K+D96L+L97M-I-Q249R SPPPRPRP+N94K+D96L+L97M+Q249R SPIRPRP+D57G-I-N94K-I-D96L+L97M+Q249R SPPRRP+I90F+D96L+E99K+V187A SPIRPRP+D137G+D167G+E21V-I-W221L E1SPIRPRP+I90F+D96L+E99K+V187A E1SRKRKRK+I90F+D96L+E99K+V187A E1SPRKPPJK+I90F+D96L+E99K-I-V187A E1SPPRRP+D62R+I90F+D96L+E99K+V187A E1SPPRRP+I90F+D96L+E99K+V187A+N200R+R209A E1SPPRRP+I90F+D96L+E99K+V187A+T199R+N200R+R209A E1SPIRPRP+D57G+D62R+N94K-I-D96L+Q249R E1SP1RPRP+D57G+N94K+D96L+N200R+R209A-I-Q249R E1SPIRPRP+D57G-I-N94K+D96L+T199R+N200R+Q249R E1SPPRRP+I90F+D96L+E99K+V187A+T199R E1SPIRPRP+D57G+N94K+D96L+T199R+R209A+Q249R E1SPIRPRP-I-I90F+D96L+E99K+V187A+Q249R E1SPPRRP+I90F+D96L+E99K+V187A+P253R E1SPPRRP+I90F+D96L+E99K+D137G+D167G+V187A+Q249R E1SPPRRP+I90F+D96L+E99K+D137G+V187A+Q249R E1SPPRRP+D96L+E99K+V187A+Q249R E1SPPRPR+V2P+N94K+D96L+Q249R E1SPPWWP+V2W+S3R+N94K+D96L+Q249R E1SPPWRP+V2R+S3R+N94K+D96L+Q249R E1SPPRWP+V2R+S3R+N94K+D96L+Q249R E1SPPWWP+V2R+S3W+N94K+D96L+Q249R E1SPPRWP+V2W+S3R+N94K+D96L+Q249R E1SPPRWP+V2R+S3W+N94K+D96L+Q249R E1SPPRWP+N94K+D96L+Q249R E1SPPRRP+N94K+D96L+Q249R E1APPPRPRPRPRP+ V2G+S3T+D57G+N94K+D96L+L97M+Q249R E1APPPRTRPRPRS+V2G+S3T+Q4P+D5E+D57G+N94K+D96L+L97M-I-Q249R E1APPPKASPRQRP+V2G+D5Q+L6M+D57G+N94K+D96L+L97M+Q249R SCIRR-I-N94K-I-D96L+E239C+Q249R E1SPPRRP+D57G+N94K+D96L+Y53C+K127C+Q249R E1SPPRRPR+V2R+S3P+N94K+D96L+Q249R E1SPPWPRP+V2R+S3P+N94K+D96L+Q249R E1SPPRRP+N94K+D96L+E99K E1SPPRRP+N94K+D96L+E99K+Q249R E1SPPCGRRP+N94K+D96L+E239C+Q249R E1SPCRPRP+N94K+D96L+E239C+Q249R SPPCRRRP+N94K+D96L+E239C+Q249R and the variants disclosed in the Examples hereinafter. When an N-terminal extension is present in the above specified variants the nomenclature "El...." is intended to indicate that the E in position 1 has been replaced by the last amino acid residue of the peptide addition Usted after "El", the remaining residues having been fused to the amino acid residue occupying position 1. For instance, "EISPPEQP" is intended to indicate that amino acid residue El has been replaced by "P" and that the remaining residues "SPPEQ" have been fused to EIP. In practice, such variants are conveniently constmcted by replacing amino acid residues (-5)-(-l) of the unprocessed parent enzyme with the relevant amino acid residues of the peptide extension and replacing El with the relevant amino acid residue, the replacements being performed by introducing the relevant mutations in the corresponding DNA sequence, and subsequently produce the resulting variant in an expression system aUowing at least a portion of the expressed variants to maintain their N-terminal extension (as is further disclosed herein). Where no replacements of El are indicated the N-terminal peptide addition is simply fused to the amino acid residue occupying position 1 of the mature parent enzyme. The above variants have initiaUy been constmcted using the random mutagenesis and/or gene shuffling methods of the invention and subsequently characterized with respect to the thereby introduced mutations. It will be apparent that an altemative method of constmcting these variants would be based on site-directed mutagenesis using suitable ohgonucleotide probes in accordance with methods known in the art. It is contemplated that the good washing performance/first wash activity wiU be maintained when one of the above specific individual mutations is replaced by a mutation to an amino acid residue belonging to the same charge group as the above suggested mutation. For instance, the mutation N94K may be replaced by N94R, H, the "G" in mutation D137G or D167G may be replaced by one of the other amino acid residues belonging to the neutral group, etc. Furthermore, it may be advantageous to replace an amino acid residue of the neutral group with one belonging to the positive charge group, e.g. the "G" in D137G and D167G, resjpectively, may be replaced with a K, R or H resulting in the mutations D137K,R,H and D167K, R,H, respectively. As already mentioned the H. lanuginosa Upolytic enzyme is structuraUy closely related to other Upolytic enzymes such as those derivable from Rhizomucor miehei, Penicillium camembertii, Absidia sp. and the various Rhizopus sp. disclosed herein. Accordingly, it is beUeved that modifications conesponding to those mentioned above in H. lanuginosa Upase and introduced into homologous positions in other structuraUy related Upolytic enzyme wiU also be functional with respect to the first wash performance. Accordingly, in a further aspect the invention relates to a first wash variant of a parent Upolytic enzyme which has an amino acid sequence or a three-dimensional stmcture which can be aUgned with the H. lanuginosa Upase amino acid sequence or stmcture (with e.g. at least 20% sequence identity aUowing gaps or an overaU protein similarity of at least 50% , such as at leas 60% or 70% using the UWGGG GAP programme or a "stmctural" similarity), the amino acid sequence of which has been modified so as to introduce mutations conesponding to those of the H. lanuginosa Upase mentioned above and/or to introduce amino acid residues found in the wUdtype H. lanuginsa Upase into the parent Upolytic enzyme in question.The amino acid residues or positions to be modified in the structuraUy or sequence homologous Upases may be identified from an aUgnment of the relevant structure/sequence with that of the H. lanuginosa Upase. Such variants may be constmcted by a method of constmcting a first wash Upolytic enzyme variant prepared from a parent Upolytic enzyme exhibiting stmctural and/or sequence homology to the H. lanuginosa Upase (such Upolytic enzymes being identified above), which method comprises ahgning the sequence of the parent enzyme in question with that of the H. lanuginosa Upase or a first wash variant thereof or superimposing the stmcture of the parent enzyme in question with that of the H. lanuginosa Upase or variant, identifymg the position(s) in the parent enzyme which are homologous to position(s) of the H. lanuginosa Upase or variant beUeved to be essential for achieving first wash activity (cf the mutations disclosed above), and replacing the amino acid residue occupying the relevant position(s) according to t, and producmg the resulting variant enzyme. Cloning a DNA sequence encoding a parent lipolytic enzyme The DNA sequence encoding a parent Upolytic enzyme from which a first wash Upolytic enzyme is created in accordance with the present invention may be isolated from any cell or microorganism producmg the parent enzyme in question by use of methods known in the art. For instance, the DNA sequence may be isolated by estabUshing a cDNA or genomic Ubrary from an organism expected to harbour the sequence, and screening for positive clones by conventional procedures. Examples of such procedures are hybridization to oUgonucleotide probes prepared on the basis of the amino acid or DNA sequence of the parent enzyme (if sequence information is avaUable) or of a related Upolytic enzyme (if sequence information as to the parent enzyme is not available) in accordance with standard techniques (cf . Sambrook et al., 1989), and/or selection for clones expressing Upolytic activity, and/or selection for clones producing a protein which is reactive with an antibody raised against a parent Upolytic enzyme. A preferred method of isolating a DNA sequence encoding a parent Upolytic enzyme to be modified in accordance with the invention from a cDNA or genomic Ubrary is by use of polymerase chain reaction (PCR) using degenerate oUgonucleotide probes prepared on the basis of DNA or amino acid sequence of the parent enzyme. For instance, the PCR may be carried out using the techniques described in US Patent No. 4,683,202 or by R.K. Saiki et al. (1988). Altematively, the DNA sequence encoding the parent enzyme may be prepared syn¬ theticaUy by estabUshed standard methods, e.g. the phosphoamidite method described by Beaucage and Caruthers (1981), or the method described by Matthes et al. (1984). According to the phosphoamidite method, oUgonucleotides are synthesized, e.g. in an automatic DNA synthesizer, purified, annealed, tigated and cloned in appropriate vectors. FinaUy, the DNA sequence encoding the parent enzyme may be prepared from DNA of mixed genomic and synthetic, mixed synthetic and cDNA or mixed genomic and cDNA origin prepared by Ugating fragments of synthetic, genomic or cDNA origin (as appropriate), the fragments conesponding to various parts of the entire DNA sequence encoding the parent enzyme, in accordance with standard techniques. Methods of constructing of first wash lipolytic enzyme variants As wiU be apparent from the brief description of the invention the present inventors have developed a very efficient method for creating Upolytic enzymes capable of removmg a substantial amount of fatty matter during a one wash cycle assay as described herein. Thus, in one highly prefened embodiment the Upolytic enzyme of the invention is a variant of a naturaUy-occuning parent Upolytic enzyme which is the result of a process comprising at least the following steps: (a) expressing a variety of mutated DNA sequences originating from a parent Upolytic enzyme in suitable host ceUs; (b) screening for host ceUs expressing a mutated Upolytic enzyme which has a decreased dependence on calcium and/or an improved tolerance towards a detergent or a detergent component as compared to the parent Upolytic enzyme; and (c) selecting a mutated Upolytic enzyme among those resulting from step (b) which, when present in detergent composition A or B in a concentration of 12500 LU/1, is capable of removing at least 15% more lard from a lard stained swatch, than the same detergent composition without the enzyme, in a one cycle wash assay as described herein. The variety of mutated DNA sequences refened to in step (a) may conveniently be obtained by subjecting a DNA sequence encoding the parent Upolytic enzyme to mutagenesis to form mutated DNA sequences Although the mutagenesis may be performed by any suitable method, such as by site-directed mutagenesis, it is presently prefened that the mutagenesis is carried out as a random mutagenesis. Thus, by use of random mutagenesis it is possible to create a much higher number of mutated DNA sequences than would be possible by use of site-directed mutagenesis. The random mutagenesis is explained in further detaU below in the section entitled "Random mutagenesis". In that section it is also described how one or more of the steps (a)-(c) of the method may be repeated one or more times in order to make successive improvements. For instance, the mutated Upolytic enzyme selected from the first round of steps (a)-(c) is subjected to a second round of the method in which the screening step (b) involves selection at more stringent conditions than those used in the screening step (b) of the first round thereby selecting for mutated Upolytic enzymes which has a decreased calcium dependence and/or an improved tolerance towards a detergent or a detergent component as compared to the mutated Upolytic enzyme resulting from the first round. Random mutagenesis The random mutagenesis of the DNA sequence encoding the parent Upolytic enzyme (or the peptide addition) to be performed in accordance with step a) of the above method may conveniently be performed by use of any method known in the art. For instance, the random mutagenesis may be performed by use of a suitable physical or chemical mutagenizing agent, by use of a suitable oUgonucleotide, or by subjecting the DNA sequence to PCR generated mutagenesis. Furthermore, the random mutagenesis may be performed by use of any combination of these mutagenizing agents. The mutagenizing agent may, e.g., be one which induces transitions, transversions, inversions, scrambling, deletions, and/or insertions. Examples of a physical or chemical mutagenizing agent suitable for the present purpose includes ultraviolet (UV) irradiation, hydroxylamine, N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), O-methyl hydroxylamine, nitrous acid, ethyl methane sulphonate (EMS), sodium bisulphite, formic acid, gamma inadiation, l-methyl-3-nitro-l-nitrosoguanidine (NTG), and nucleotide analogues. When such agents are used the mutagenesis is typicaUy performed by incubating the DNA sequence encoding the parent enzyme to be mutagenized in the presence of the mutagenizing agent of choice under suitable conditions for the mutagenesis to take place, and selecting for mutated DNA having the desired properties. When the mutagenesis is performed by the use of an oUgonucleotide, the oUgonucleotide may be doped or spiked with the three non-parent nucleotides during the synthesis of the oUgonucleotide at the positions wanted to be changed. The doping or spiking may be done so that codons for unwanted amino acids are avoided by lowering the amount of or completely avoiding the nucleotides resulting in these codons. State of the art knowledge and computer programs can be used for calculating the most optimal nucleotide mixture for a given amino acid preference. The doped or spiked oUgonucleotide can be incorporated into the DNA encoding the Upolytic enzyme by any pubUshed technique using e.g. PCR, LCR or any DNA polymerase and Ugase. When PCR generated mutagenesis is used either a chemicaUy treated or non-treated gene encoding a parent Upolytic enzyme is subjected to PCR under conditions that increases the mismcorporation of nucleotides (Deshler 1992, Leung et al. 1989). A mutator strain of E. coli (Fowler et al. 1974), S. cerevisiae or any other microbial organism may be used for the random mutagenesis of the DNA encoding the Upolytic enzyme by e.g. transforming a plasmid containing the parent enzyme into the mutator strain, growing the mutator strain with the plasmid and isolating the mutated plasmid from the mutator strain. The mutated plasmid may subsequently be transformed into the expression organism. The DNA sequence to be mutagenized may conveniently be present in a genomic or cDNA Ubrary prepared from an organism expressing the parent Upolytic enzyme. Altematively, the DNA sequence may be present on a suitable vector such as a plasmid or a bacteriophage, which as such may be incubated with or otherwise exposed to the mutagenizing agent. The DNA to be mutagenized may also be present in a host ceU either by being integrated in the genome of said ceU or by being present on a vector harboured in the ceU. FinaUy, the DNA to be mutagenized may be in isolated form. The DNA sequence to be subjected to random mutagenesis is preferably a cDNA or a genomic DNA sequence. In some cases it may be convenient to ampUfy the mutated DNA sequence prior to the expression or screening being performed. Such ampUfication may be performed in accordance with methods known in the art, the presently prefened method being PCR generated amplification using oUgonucleotide primers prepared on the basis of the DNA or amino acid sequence of the parent enzyme. Subsequent to the incubation with or exposure to the mutagenizing agent, the mutated DNA is expressed by culturing a suitable host ceU carrying the DNA sequence under conditions aUowing expression to take place. The host ceU used for this purpose may be one which has been transformed with the mutated DNA sequence, optionaUy present on a vector, or one which carried the DNA sequence encoding the parent enzyme during the mutagenesis treatment. Examples of suitable host ceUs are given below. It is particularly prefened to use a yeast ceU as a host ceU, in particular when the parent Upolytic enzyme is derived from a fungus such as a filamentous fungus or yeast. The mutated DNA sequence may further comprise a DNA sequence encoding functions permitting expression of the mutated DNA sequence. It will be understood that the screening criteria mentioned in step (b) of the method of the invention have been carefuUy selected. Thus, without being Umited to any theory the screening for a decreased dependency on calcium at alkaline pH (pH above 7) is beUeved to result in variants having an over-aU improved performance in that the requirement for calcium may be considered a Umiting factor for optimal activity, in particular under most wash conditions which are caracterized by the fact that the concentration of free calcium ions is deUberately lowered by chelating agents in the detergent matrix (buUders). The detergent or detergent component towards which the variant has improved tolerance may be of any type, e.g. as further described below. Preferably, the detergent component is a non-ionic, anionic, cationic, zwitterionic or amphoteric surfactant. Examples of non-ionic surfactants include an alcohol ethoxylate, examples of anionic surfactants include LAS, alkyl sulphate, alcohol ethoxy sulphate and the like. The choice of detergent wiU, e.g., depend on the inherent weakness (in relation to detergent tolerances) of the parent Upolytic enzyme. In relation Humicola lanuginosa Upolytic enzymes and homologous enzymes (such as the Penicillium, Rhizomucor, Rhizopus and Absidia sp. Upolytic enzymes), it is contemplated that an improved tolerance towards a non-ionic surfactant alcohol ethoxylate, a commerciaUy avaUable example of which is Dobanol ® 25-7, may be indicative of improved wash performance. In relation to Pseudomonas type Upolytic enzymes such as P. pseudoalcaligenes, P. cepacia, it is contemplated that an improved tolerance towards an anionic surfactant such as an alkyl sulphate (a commericaUy avaUable example of which is NEODOL 45) or LAS (a commerciaUy avaUable example of which is Nansa 1169/P) may be indicative of improved wash performance. The screening of step (b) is conveniently performed by use of a filter assay based on the foUowing principle: A microorganism capable of expressing the mutated Upolytic enzyme of interest is incubated on a suitable medium and under suitable conditions for the enzyme to be secreted, the medium being provided with a double filter comprising a first protein-binding filter and on top of that a second filter exhibiting a low protein binding capability. The microorganism is located on the second filter. Subsequent to the incubation, the first filter comprising enzymes secreted from the microorganisms is separated from the second filter comprising the micro¬ organisms. The first filter is subjected to screening for the desired enzymatic activity and the corresponding microbial colonies present on the second filter are identified. Altematively, the second filter carrying the colonies may be used directiy on the screening plate. This makes it easier to pick the right colonies and in some cases gives a stronger signal. And using only one filter, either protein binding or none-protein binding is sufficient in many cases. The filter used for binding the enzymatic activity may be any protein binding filter e.g. nylon or nitroceUulose. The topfilter carrying the colonies of the expression organism may be any filter that has no or low affinity for binding proteins e.g. ceUulose acetate or Durapore™. The filter may be pretreated with any of the conditions to be used for screening or may be treated during the detection of enzymatic activity. The enzymatic activity may be detected by a dye, fluorescence, precipitation, pH indicator, IR-absorbance or any other known technique for detection of enzymatic activity. The detecting compound may be immobilized by any immobilizing agent e.g. agarose, agar, gelatine, polyacrylamide, starch, filter paper, cloth; or any combination of immobilizing agents. Lipolytic activity may be detected by BrilUant green, Rhodamine B or Sudan Black in combination with a Upid e.g. oUve oU or lard. The screening criteria for identifying variants of parent Upolytic enzymes having improved washing performance may be e.g. EGTA, EDTA, non-ionic and/or anionic tensides, alkaline pH, or any detergent composition in combmation with one of the above detectors of enzymatic activity. Subsequent to the screening in step (b) Upolytic enzymes having desired properties (i.e. as defined by the screening criteria) are isolated and their first wash capability tested in the one cycle wash assay described in the Materials and Methods section herein. If the first wash activity of the enzyme is not sufficiently good after one round of the above treatment, the enzyme may be modified, e.g. by site-directed or random mutagenesis in order to improve the first wash activity of the enzyme, e.g. in accordance with any of the principles given further above for modifacation of Upases to achieve a first wash performance. Most convenient, the host ceUs produced in step (b) may be subjected to further rounds of mutagenesis as defined in steps (a)-(b) and optionaUy (c) above, conveniently by using more stringent selection criteria than employed in a previous mutagenesis treatment. The further round(s) of mutagenesis may be random, localized random or site-directed so as to introduce previously identified advantageous mutations, in particular D96L, Q249R, E87K, D254K, E210K or to introduce random mutations in selected regions, e.g. the Upid contact zone, in particular random mutations with doped or spiked oUgonucleotides towards introduction of positive and/or hydrophobic amino acid residues, or to introduce any of the other specific mutations mentioned herein. Altematively, genes encoding different homologous parent Upolytic enzymes may be combined in a random manner in order to obtain a novel variant carrying one or more mutations from each variant. This is discussed in further detaU below in the section entitied "Combination of DNA sequences encoding Upolytic enzymes". The host ceUs selected for in step (b) or (c) may be used directly for the production of the variant of the Upolytic enzyme. Altematively, DNA encoding the variant may be isolated from the host ceU and inserted into another suitable host ceU, conveniently by use of the procedure described below in the section entitied "Expression of a variant of the invention" , in which suitable host ceUs are also Usted. Localized random mutagenesis In accordance with the invention the random mutagenesis may advantageously be located to a part of the parent Upolytic enzyme in question. This may, e.g. , be advantageous when a certain region of the enzyme has been identified to be of particular importance for a given property of the enzyme, and which, when modified, is expected to result in a variant having improved properties. Such region may normaUy be identified when the tertiary stmcture of the parent enzyme has been elucidated and related to the function of the enzyme. One area of particular interest for modification amino acid residues located at the surface of the parent enzyme within or outside the Upid contact zone, i.e. the part of the Upolytic enzyme which is in contact with the Upid substrate and e.g. comprising the Ud region, the hydrophobic cleft or any part of these stmctures. Another area of interest for Upolytic enzymes of the invention which contains a peptide addition or another modification within a non-stmctural part of the N-terminal or C-terminal end of the mature parent enzyme. The localized random mutagenesis is conveniently performed by use of PCR generated mutagenesis techniques as described above or any other suitable technique known in the art. EspeciaUy for mutagenizing large peptide additions, it may be relevant to use PCR generated mutagenesis (e.g. as described by Deshler 1992 or Leung et al., 1989), in which one or more suitable oUgonucleotide probes are used which flanks the area to be mutagenized. For mutagenesis of shorter peptide additions, it is more preferably perform the localized random mutagenesis by use of doped or spiked oUgonucleotides. The doping or spiking is used, e.g., to avoid codons for unwanted amino acid residues or to increase the likelihood that a particu¬ lar type of amino acid residue, such as a positively charged or hydrophobic amino acid resi¬ due, is introduced at a desired position. Altematively, the DNA sequence encoding the part of the DNA sequence to be modified may be isolated, e.g. by being inserted into a suitable vector, and said part may subsequently be subjected to mutagenesis by use of any of the mutagenesis methods discussed above. Of particular interest is that the DNA sequence subjected to random mutagenesis com¬ prises a part of or constitutes a part of a DNA sequence encoding the Upid contact zone or the Ud region of the parent Upolytic enzyme. The localized random mutagenesis may be performed in one or more of these regions and/or one or more of the regions constituting the Upid con- tact zone, and is preferably performed in at least two of the regions. Parent Upolytic enzymes of particular interest for modification according to this aspect of the invention includes the H. lanuginosa Upolytic enzyme obtainable from strain DSM 4109 or a variant or analogue thereof, a parent Upolytic enzyme derived from Penicillium camembertii, a parent Upolytic enzyme derived from Rhizopus oryzae, a parent Upolytic enzyme derived from Rhizomucor miehei, a parent Upolytic enzyme derived from a Absidia sp. Upolytic enzyme, a parent Upolytic enzyme derived from a Pseudomonas sp., prefereably belonging to the Ps. aerogi- nosa famUy such as the Pseudomonas cepacia Upase, the Pseudomonas pseudoalcaligenes U- pase, the Pseudomonas glumae Upase, the Pseudomonas mendocina Upase, the Pseudomonas wisconsinensis, or the Pseudomonas sp. Upase (SD705) (Liposam ® ) shown in SEQ ID NO 99. The Upid contact zones and Ud regions are identified in the Definitions section above. The localized random mutagenesis may be done by use of doped oUgonucleotides which are doped in the direction of L, I, V, F, W, A (hydrophobic amino acid residues) or K,R (positive amino acid residues), for instance under conditions ensuring about 90-93% wUdtype and about 7-10% mutant. Specific examples of suitable doping regimes are given in the Examples section below. In vivo recombination According to a prefened embodiment of the invention a DNA sequence encoding a first wash Upolytic enzyme may be constmcted by a method, which as an important step involves combination of selected DNA sequences encoding different parent Upolytic enzymes or parts of such DNA sequences. Preferably, the DNA sequences to be combmed are derived from genes encoding Upolytic enzymes which has a satisfactory washing and/or dishwashing performance (e.g. as identified in Example 6) The aim of combining the DNA sequences is that the best elements from each "parent enzyme" are combined into one and the same variant enzyme. In the present context the term "satisfactory washing performance" is intended to indicate that the parent enzymes are capable of removing fatty stains during one or several wash cycles when present in a suitable detergent. Preferably, the parent enzyme in question has a better washing performance than Iipolase(TM). The combination of DNA sequences may be performed by any suitable method known in the art. For instance, when the DNA sequences to be combined comprises homologous frag¬ ments, the combination is preferably achieved by homologous cross-over, e.g. by use of con¬ ventional methods such as US 5, 093, 257, or by gene shuffling (Stemmer (1994), Proc. Natl. Acad. Sci. USA, vol. 91, 10747-10751; Stemmer (1994), Nature, vol. 370, 389-391; Smith (1994), Nature vol 370, page 324-25), WO 95/17413. Gene shuffling means recombination of nucleotide sequence(s) between two or more homologous DNA sequences resulting in out¬ put DNA sequences having a number of nucleotides exchanged. Of particular interest is an in vivo Gene Shuffling Method which is based on the foUow¬ ing procedure: a) forming at least one circular expression vector comprising a DNA sequence encoding a parent Upolytic enzyme or a substantial part thereof, b) opening said circular expression vector within the DNA sequence encoding the Upolytic enzyme or part thereof , c) preparing at least one DNA fragment comprising a DNA sequence homologous to at least a part of the enzyme coding region on at least one of the circular expression vector(s), d) introducing at least one of said opened vector(s), together with at least one of said homolo¬ gous DNA fragments) covering fuU-length DNA sequences encoding said Upolytic enzyme(s) or a part thereof, into a recombination host ceU, e) cultivating said yeast recombination host ceU under conditions conducive for recombination betwen the homologous DNA fragments to take place, and f ) screening for positive Upolytic enzyme variants with an improved wash performance. The vector used in step a) above may be a yeast expression vector which can be trans- formed into and expressed in a yeast recombination host ceU. Examples of such expression vectors include yeast expression vectors constmcted from pYES 2.0 (Invitrogene), such as pJSO37 comprising the wUd type Humicola lanuginosa Upase gene Opening of the vector in step b) may be accompUshed by any conventional techniques known in the art, and may for instance be performed by opening the vector within the Upase gene by cutting at a single site or by gapping the vector (i.e. cutting e.g. at two sites result¬ ing in cutting out a Uttle part of the gene). The preparation of the homologous DNA fragment(s) in step c) may be performed by amplifying homologous DNA sequence(s) (e.g. comprising one or more mutation in the Upolytic gene and comprismg in a plasmid or vector) by any suitable methods, such as by a standard PCR amplification method described in US 4,683,202 or Saiki et al., (1988), Sci¬ ence 239, 487 - 491). The vector(s) may be introduced into the recombination host ceU (in step d) by transfor¬ mation. In the case of the recombination host ceU is a strain of Saccharomyces cerevisiae, such as Saccharomyces cerevisiae YNG318 (described below) the transformation may be per¬ formed as described by Sambrooks et al., (1989), Molecular Cloning: A Laboratory Manual, 2nd Ed., Cold Spring Harbor, NY, USA). The screening for positive Upolytic enzyme variants may, e.g. , be performed by the screening method described in connection with the random mutagenesis above. One of more cycle of step a) to f) may be performed before a selection of a first wash Upolytic enzyme variant is made using the selection conditions defined further above. According to the shuffling method significantiy more than two DNA sequences can be shuffled. Any number of different DNA fragments and homologous Upolytic enzymes com¬ prised in suitable plasmids may be shuffles at the same time. The DNA sequences to be combmed may be entire genes of which at least one part exhibits sufficient homology to the other genes to aUow for recombination of the genes to take place. Altematively, the DNA sequences may be partial genes, which when combined can give rise to a functional gene capable of expressing a Upolytic enzyme. When the DNA sequences to be combined are highly homologous or partiaUy identical controUed combmation may be performed, e.g. in the case of combmation of two DNA sequences, to combine the N-terminal part of one of the sequences with the C-terminal part of the other (conesponding to the remaining part of the first sequence) or by combining other relevant parts of the respective genes in question. NaturaUy occurring enzymes may be geneticaUy modified by random, localized random or site directed mutagenesis as described above prior to bemg subjected to gene shuffling. Al¬ tematively, part of one enzyme may be replaced by a part of another to obtain a chimeric enzyme. This replacement can be achieved either by conventional in vitro gene spUcing tech¬ niques or by in vivo recombination or by combinations of both techniques. When using conventional in vitro gene spUcing techniques, a desired portion of the Upolytic enzyme gene may be deleted using appropriate site-specific restriction enzymes; the deleted portion ofthe coding sequence may then be replaced by insertion of a desired portion of a different Upolytic enzyme coding sequence so that a chimeric nucleotide sequence encoding a new Upolytic enzyme is produced. Altematively, Upolytic enzyme genes may be fused, e.g. by use of the PCR overlay addition method described by Higuchi et al. 1988. The in vivo recombination techniques depend on the fact that different DNA segments with highly homologous regions (identity of DNA sequence) may recombine, i.e. break and exchange DNA, and estabUsh new bonds in the homologous regions. Accordingly, when the coding sequences for two or more different but homologous Upolytic enzymes are used to transform a host ceU, recombination of homologous sequences in vivo w l result in the production of chimeric gene sequences. Translation of these coding sequences by the host ceU wUl result in production of a chimeric Upolytic enzyme gene product. Specific in vivo recombination techniques are described in US 5,093,257 and EP 252 666. In order to aUow homologous recombination to take place it is desirable that the Upolytic enzymes comprises parts which are at least 60% homologous. It is particularly preferred that the entire enzymes are at least 60% homologous. The enzymes to be combmed may be different variants of the same parent enzyme, e.g. variants derived from the H. lanuginosa Upolytic enzyme disclosed herein, or variants derived from the Ps. alcaligenes or Ps. pseudoalcaligenes Upolytic enzymes refened to further above, or variants derived from the F. solanipisi Upolytic enzyme (cf above), or variants derived from the P. mendocina Upolytic enzyme or the Pseudomonas sp. Upase (Liposam) (cf. above). It will be understood that the random recombination may be performed between a naturaUy-occuning Upolytic enzymes and one or more variants of said enzyme, between differerent naturaUy ocurring enzymes, between variants of naturaUy ocurring enzymes (the variants being variants of the same parent enzyme or of different enzymes), or between any combination of naturaUy occurring enzymes and variants of naturaUy occurring enzyme as long as the corresponding DNA sequences are capable of recombinng. When the DNA sequences to be combined are variants of a parent enzyme these variants may conveniently be prepared by the mutagenesis, in particular random mutagenesis method disclosed above. In an altemative embodiment, the hybrid enzyme may be synthesized by standard chemical methods known in the art. For example, see ΗunkapiUer et al. (1984). Accordingly, peptides having the amino acid sequences described above may be synthesized in whole or in part and joined to form the hybrid enzymes of the invention. In a highly prefened embodiment first wash Upolytic enzymes of the invention are constmcted by a method which comprises subjecting a parent Upolytic enzyme to mutagenesis, in particular random mutagenesis, to form a variety of mutated DNA sequences, expressing the mutated DNA sequences in a suitable host and screening for host ceUs which produces a mutated Upolytic enzyme which has a decreased dependency on calcium and/or an improved tolerance towards a detergent or a detergent component, subjecting the DNA sequence encoding the mutated Upolytic enzyme selected in said screening to in vivo recombination, in particular gene shuffling or sexual PCR, with one or more other mutated DNA sequences prepared in a similar manner from the same parent Upolytic enzyme, expressing the mutated recombined DNA sequences in a suitable host, optionaUy selecting for host ceUs producing a mutated Upolytic enzyme which has a decreased dependency on calcium and/or an improved tolerance towards a detergent or a detergent component, optionaUy repeating either or both of the above mutagenesis and in vivo recombination procedures one or more times using more stringent screening criteria, and finaUy selecting a recombined DNA sequence endoding a Upolytic enzyme exhibiting first wash activity as defined herein. Furthermore, it will be understood that a first wash Upolytic enzyme of the invention which comprises a peptide addition as weU as mutation(s) in a stmctural part of the parent enzyme may be constmcted by a method which involves localized mutagenesis, in particular localized random mutagenesis, in the part of the DNA sequence encoding the peptide addition and selected parts of the DNA sequence encoding the mature part of the parent Upolytic enzyme, i.e. a combination ofthe random mutagenesis method according to the second aspect of the invention performed in a stmctural part of the parent enzyme and random mutagenesis in a non-structural part of the N-terminal and/or C-terminal end and/or in a peptide addition appUed to the N-terminal and/or C-terminal part. It will be understood that the in vivo recombination and mutagenesis methods disclosed herein may be appUed to any of the parent Upolytic enzymes mentioned in the "Parent Lipolytic Enzymes" section herem. Particularly prefened parent Upolytic enzymes are derived from Humicola lanuginosa and from Pseudomonas sp. such as Ps. alcaligenes and Ps. pseudoalcaligenes. Expression of a lipolytic enzyme of the invention An isolated nucleic acid sequence encoding a first wash Upolytic enzyme of the invention may be manipulated in a variety of ways to provide for expression of the enzyme. Manipulation of the nucleic acid sequence encoding a first wash Upolytic enzyme prior to its insertion into a vector may be desirable or necessary depending on the expression vector. The techniques for modifying nucleic acid sequences utilizing cloning methods are weU known in the art. The term "control sequences" is defined herein to include aU components which are necessary or advantageous for expression of the coding sequence of the nucleic acid sequence. Each control sequence may be native or foreign to the nucleic acid sequence encoding the Upolytic enzyme. Such control sequences include, but are not Umited to, a leader, a polyadenylation sequence, a propeptide sequence, a promoter, a signal sequence, and a transcription terminator. At a minimum, the control sequences include a promoter, and transcriptional and translational stop signals. The control sequences may be provided with linkers for the purpose of introducing specific restriction sites facilitating Ugation of the control sequences with the coding region of the nucleic acid sequence encoding the Upolytic enzyme. The control sequence may be an appropriate promoter sequence, a nucleic acid sequence which is recognized by a host ceU for expression of the nucleic acid sequence. The promoter sequence contains transcription and translation control sequences which mediate the expression of the first wash Upolytic enzyme. The promoter may be any nucleic acid sequence which shows transcriptional activity in the host ceU of choice and may be obtained from genes encoding extraceUular or intraceUular polypeptides either homologous or heterologous to the host ceU. Examples of suitable promoters for directing the transcription of the nucleic acid constmcts of the present invention, especiaUy in a bacterial host ceU, are the promoters obtained from the E. coli lac operon, the Streptomyces coelicolor agarase gene (dagA), the B. subtilis levansucrase gene (sacB) or the alkaline protease gene, the B. licheniformis alpha-amylase gene (amyL), the B. stearothermophilus maltogenic amylase gene (amyM), the B. amyloliquefaciens alpha-amylase gene (amyQ), the B.licheniformis penicilUnase gene (penP), the B. subtilis xylA and xylB genes, the B. pumilus xylosidase gene, and the prokaryotic beta-lactamase or tryptophan gene (ViUa-Kamaroff et al. , 1978, Proceedings ofthe National Academy of Sciences USA 75:3727-3731), as weU as the tac gene (DeBoer et al. , 1983, Proceedings ofthe National Academy of Sciences USA 80:21-25). Further promoters are described in "Useful proteins from recombinant bacteria" in Scientific American, 1980, 242:74-94; and in Sambrook et al. , 1989, supra. Examples of suitable promoters for directing the transcription of the nucleic acid constmcts of the present invention in a filamentous fungal host ceU are promoters obtained from the genes encoding A. oryzae TAKA amylase, A. oryzae triose phosphate isomerase, Rhizomucor miehei aspartic proteinase, A. niger neutral alpha-amylase, A. niger acid stable alpha-amylase, A. niger or A..awamori glucoamylase (glaA), Rhizomucor miehei Upase, A. oryzae alkaline protease, A. oryzae triose phosphate isomerase, A. nidulans acetamidase, Fusarium oxysporum trypsin-like protease (as described in U.S. Patent No. 4,288,627, which is incorporated herein by reference), or the ADH-3 promoter (McKnight et al., (1985), The EMBO J. 4, 2093-3099) and hybrids thereof. Particularly prefened promoters for use in filamentous fungal host ceUs is the TAKA amylase and the glaA promoters. In a yeast host, promoters from yeast glycolytic genes (Hitzeman et al.,(1980), J. Biol. " Chem. 255, 12073-12080; Alber and Kawasaki, (1982), J. Mol. Appl. Gen. 1, 419-434) or alcohol dehydrogenase genes (Young et al., in Genetic Engmeering of Microorganisms for Chemicals (HoUaender et al, eds.), Plenum Press, New York, 1982), or the TPI1 (US 4,599,311) or ADH2-4c (RusseU et al., (1983), Nature 304, 652-654) promo¬ ters, useful promoters are obtained from the S. cerevisiae enolase (ENO-1) gene, the S. cerevisiae galactokinase gene (GAL1), the S. cerevisiae alcohol dehydrogenase/glyceraldehyde-3-phosphate dehydrogenase genes (ADH2/GAP), and the S. cerevisiae 3-phosphoglycerate kinase gene. Other useful promoters for yeast host ceUs are described by Romanos et al. , 1992, Yeast 8:423-488. The control sequence may also be a suitable transcription terminator sequence, a sequence recognized by a host ceU to terminate transcription. The terminator sequence is operably linked to the 3' terminus of the nucleic acid sequence encoding the modified Upolytic enzyme. The terminator sequence may be native to the nucleic acid sequence encoding the Upolytic enzyme or may be obtained from foreign sources. Any terminator which is functional in the host ceU of choice may be used in the present invention. Preferred terminators for filamentous fungal host ceUs are obtained from the genes encoding A. oryzae TAKA amylase, A. niger glucoamylase, A. nidulans anthranilate synthase, A. niger alpha-glucosidase, and Fusarium oxysporum trypsin-like protease. Prefened terminators for yeast host ceUs are obtained from the genes encoding S. cerevisiae enolase, S. cerevisiae cytochrome C (CYC1), or S. cerevisiae glyceraldehyde-3-phosphate dehydrogenase. Other useful te-rninators for yeast host ceUs are described by Romanos et al. , 1992, supra. The control sequence may also be a suitable leader sequence, a nontranslated region of a mRNA which is important for translation by the host ceU. The leader sequence is operably linked to the 5' terminus of the nucleic acid sequence encoding the Upolytic enzyme. The leader sequence may be native to the nucleic acid sequence encoding the Upolytic enzyme or may be obtained from foreign sources. Any leader sequence which is functional in the host ceU of choice may be used in the present invention. Prefened leaders for filamentous fungal host ceUs are obtained from the genes encoding A. oryzae TAKA amylase and A. oryzae triose phosphate isomerase. Suitable leaders for yeast host ceUs are obtained from the S. cerevisiae enolase (ENO-1) gene, the S. cerevisiae 3-phosphoglycerate kinase gene, the S. cerevisiae alpha-factor, and the S. cerevisiae alcohol dehydrogenase/glyceraldehyde-3-phosphate dehydrogenase genes (ADH2/GAP). The control sequence may also be a polyadenylation sequence, a sequence which is operably linked to the 3' terminus of the nucleic acid sequence and which, when transcribed, is recognized by the host ceU as a signal to add polyadenosine residues to transcribed mRNA. The polyadenylation sequence may be native to the nucleic acid sequence encodmg the Upolytic enzyme or may be obtained from foreign sources. Any polyadenylation sequence which is functional in the host ceU of choice may be used in the present invention. Preferred polyadenylation sequences for filamentous fungal host ceUs are obtained from the genes encoding A. oryzae TAKA amylase, A. niger glucoamylase, A. nidulans anthranilate synthase, and A. niger alpha-glucosidase. Useful polyadenylation sequences for yeast host cells are described by Guo and Sherman, 1995, Molecular Cellular Biology 15:5983-5990. Polyadenylation sequences are weU known in the art for mammaUan host ceUs. The control sequence may also be a signal peptide coding region, which codes for an amino acid sequence linked to the amino terminus of the first wash Upolytic enzyme which can direct the expressed first wash Upolytic enzyme into the ceU's secretory pathway. The signal peptide coding region may be native to the first wash Upolytic enzyme of the invention or may be obtained from foreign sources. The 5' end of the coding sequence of the nucleic acid sequence may inherently contain a signal peptide coding region naturaUy linked in translation reading frame with the segment of the coding region which encodes the secreted first wash Upolytic enzyme. Altematively, the 5' end of the coding sequence may contain a signal peptide coding region which is foreign to that portion of the coding sequence which encodes the secreted first wash Upolytic enzyme. The foreign signal peptide coding region may be required where the coding sequence does not normaUy contain a signal peptide coding region. Altematively, the foreign signal peptide coding region may simply replace the natural signal peptide coding region in order to obtain enhanced secretion of the Upolytic enzyme relative to the natural signal peptide coding region normaUy associated with the coding sequence. The signal peptide coding region may be obtained from a glucoamylase or an amylase gene from an Aspergillus species, a Upase or proteinase gene from a Rhizomucor species, the gene for the a-factor from Saccharomyces cerevisiae, an amylase or a protease gene from a Bacillus species, or the calf preprochymosin gene. An effective signal peptide coding region for bacterial host ceUs is the signal peptide coding region obtained from the maltogenic amylase gene from Bacillus NCIB 11837, the B. stearothermophilus alpha- amylase gene, the B. licheniformis subtilisin gene, the B. licheniformis beta-lactamase gene, the B. stearothermophilus neutral proteases genes (nprT, nprS, nprM), and the B. subtilis PrsA gene. Further signal peptides are described by Simonen and Palva, 1993, Microbiological Reviews 57: 109-137. An effective signal peptide coding region for filamentous fungal host cells is the signal peptide coding region obtained from A. oryzae TAKA amylase gene, A. niger neutral amylase gene, the Rhizomucor miehei aspartic proteinase gene, the H. lanuginosa ceUulase gene, or the Rhizomucor miehei Upase gene. Useful signal peptides for yeast host ceUs are obtained from the genes for S. cerevisiae a- factor and S. cerevisiae invertase. Other useful signal peptide coding regions are described by Romanos et al. , 1992, supra. However, any signal peptide coding region capable of directing the expressed enzyme into the secretory pathway of a host ceU of choice may be used in the present invention. The nucleic acid constmcts of the present invention may also comprise one or more nucleic acid sequences which encode one or more factors that are advantageous in the expression of the first wash Upolytic enzyme, e.g. , an activator (e.g. , a trans-acting factor), a chaperone, and a processing protease. The nucleic acids encoding one or more of these factors are not necessarily in tandem with the nucleic acid sequence encoding the first wash Upolytic enzyme. An activator is a protein which activates transcription of a nucleic acid sequence encoding a first wash Upolytic enzyme (Kudla et al. , 1990, EMBO Journal 9: 1355- 1364; Jarai and Buxton, 1994, Current Genetics 26:2238-244; Verdier, 1990, Yeast 6:271- 297). The nucleic acid sequence encoding an activator may be obtained from the genes encoding B. stearothermophilus NprA (nprA), S. cerevisiae heme activator protein 1 (hapl), S. cerevisiae galactose metabolizing protem 4 (gal4), and A. nidulans ammonia regulation protein (areA). For further examples, see Verdier, 1990, supra and MacKenzie et al. , 1993, Journal of General Microbiology 139:2295-2307. A chaperone is a protein which assists another polypeptide in folding properly (Harti et al. , 1994, 77-SS 19:20-25; Bergeron et al. , 1994, ΩBS 19:124-128; Demolder et al., 1994, Journal of Biotechnology 32:179-189; Craig, 1993, Science 260:1902-1903; Gething and Sambrook, 1992, Nature 355:33-45; Puig and GUbert, 1994, Journal of Biological Chemistry 269:7764-7771; Wang and Tsou, 1993, The FASEB Journal 1: 1515-11157; Robinson et al., 1994, Bio/Technology 1:381-384). The nucleic acid sequence encoding a chaperone may be obtained from the genes encoding B. subtilis GroE proteins, A. oryzae protein disulphide isomerase, S. cerevisiae calnexin, S. cerevisiae BiP/GRP78, and S. cerevisiae Hsp70. For further examples, see Gething and Sambrook, 1992, supra, and Harti et al. , 1994, supra. Any factor that is functional in the host ceU of choice may be used in the present invention. It may also be desirable to add regulatory sequences which aUow the regulation of the expression of the first wash Upolytic enzyme relative to the growth of the host ceU. Examples of regulatory systems are those which cause the expression of the gene to be tumed on or off in response to a chemical or physical stimulus, including the presence of a regulatory compound. Regulatory systems in prokaryotic systems would include the lac, tac, and trp operator systems. In yeast, the ADH2 system or GAL1 system may be used. In filamentous fungi, the TAKA alpha-amylase promoter, A. niger glucoamylase promoter, and the A oryzae glucoamylase promoter may be used as regulatory sequences. Other examples of regulatory sequences are those which aUow for gene ampUfication. In eukaryotic systems, these include the dihydrofolate reductase gene which is ampUfied in the presence of methotrexate, and the metaUothionein genes which are amplified with heavy metals. In these cases, the nucleic acid sequence encoding the first wash Upolytic enzyme would be placed in tandem with the regulatory sequence. Expression Vectors The present invention also relates to recombinant expression vectors comprising a nucleic acid sequence of the present invention, a promoter, and transcriptional and translational stop signals. The various nucleic acid and control sequences described above may be joined together to produce a recombinant expression vector which may include one or more convenient restriction sites to aUow for insertion or substitution of the nucleic acid sequence encoding the first wash Upolytic enzyme at such sites. Altematively, the nucleic acid sequence of the present invention may be expressed by inserting the nucleic acid sequence or a nucleic acid constmct comprismg the sequence into an appropriate vector for expression. In creating the expression vector, the coding sequence is located in the vector so that the coding sequence is operably linked with the appropriate control sequences for expression, and possibly secretion. The recombinant expression vector may be any vector which can be conveniently subjected to recombinant DNA procedures and can bring about the expression of the nucleic acid sequence. The choice of the vector wiU typicaUy depend on the compatibility of the vector with the host ceU into which the vector is to be introduced. The vectors may be linear or closed circular plasmids. The vector may be an autonomously repUcating vector, i.e. , a vector which exists as an extrachromosomal entity, the repUcation of which is independent of chromosomal repUcation, e.g. , a plasmid, an extrachromosomal element, a minichromosome, or an artificial chromosome. The vector may contain any means for assuring self-repUcation. Altematively, the vector may be one which, when introduced into the host ceU, is integrated into the genome and repUcated together with the chromosome(s) into which it has been integrated. The vector system may be a single vector or plasmid or two or more vectors or plasmids which together contain the total DNA to be introduced into the genome of the host ceU, or a transposon. The vectors of the present invention preferably contain one or more selectable markers which permit easy selection of transformed ceUs. A selectable marker is a gene the product of which provides for biocide or viral resistance, resistance to heavy metals, prototrophy to auxotrophs, and the like. Examples of bacterial selectable markers are the dal genes from B. subtilis or B. licheniformis, or markers which confer antibiotic resistance such as ampicillin, kanamycin, chloramphenicol or tetracycline resistance. A frequently used mammaUan marker is the dihydrofolate reductase gene. Suitable markers for yeast host ceUs are ADE2, HIS3, LEU2, LYS2, MET3, TRPl, and URA3. A selectable marker for use in a filamentous fungal host ceU may be selected from the group including, but not Umited to, amdS (acetamidase), argB (ornithine carbamoyltransferase), bar (phosphinothricin acetyltransferase), hygB (hygromycin phosphotransferase), niaD (nitrate reductase), pyrG (orotidine-5' -phosphate decarboxylase), sC (sulfate adenyltransferase), trpC (anthranilate synthase), and glufosinate resistance markers, as weU as equivalents from other species. Prefened for use in an Aspergillus ceU are the amdS and pyrG markers of A. nidulans or A. oryzae and the bar marker of Streptomyces hygroscopicus. Furthermore, selection may be accompUshed by co-transformation, e.g. , as described in WO 91/17243, where the selectable marker is on a separate vector. The vectors of the present invention preferably contain an element(s) that permits stable integration of the vector into the host ceU genome or autonomous repUcation of the vector in the ceU independent of the genome of the ceU. The vectors of the present invention may be integrated into the host ceU genome when introduced into a host ceU. For integration, the vector may rely on the nucleic acid sequence encoding the first wash Upolytic enzyme or any other element of the vector for stable integration of the vector into the genome by homologous or nonhomologous recombination. Altematively, the vector may contain additional nucleic acid sequences for directing integration by homologous recombination into the genome of the host ceU. The additional nucleic acid sequences enable the vector to be integrated into the host cell genome at a precise location(s) in the chromosome(s). To increase the likelihood of integration at a precise location, the integrational elements should preferably contain a sufficient number of nucleic acids, such as 100 to 1,500 base pairs, preferably 400 to 1,500 base pairs, and most preferably 800 to 1,500 base pairs, which are highly homologous with the conesponding target sequence to enhance the probability of homologous recombination. The integrational elements may be any sequence that is homologous with the target sequence in the genome of the host ceU. Furthermore, the integrational elements may be non-encoding or encoding nucleic acid sequences. On the other hand, the vector may be integrated into the genome of the host ceU by non-homologous recombination. These nucleic acid sequences may be any sequence that is homologous with a target sequence in the genome of the host ceU, and, furthermore, may be non-encoding or encoding sequences. For autonomous repUcation, the vector may further comprise an origin of repUcation enabling the vector to repUcate autonomously in the host ceU in question. Examples of bacterial origins of repUcation are the origins of repUcation of plasmids pBR322, pUC19, pACYC177, pACYC184, pUBHO, pE194, pTA1060, and pAMBl. Examples of origin of repUcations for use in a yeast host ceU are the 2 micron origin of repUcation, the combination of CEN6 and ARS4, and the combination of CEN3 and ARS1. The origin of repUcation may be one having a mutation which makes its functioning temperature-sensitive in the host ceU (see, e.g., EhrUch, 1978, Proceedings ofthe National Academy of Sciences USA 75:1433). More than one copy of a nucleic acid sequence encoding a first wash Upolytic enzyme of the present invention may be inserted into the host ceU to amptify expression of the nucleic acid sequence. Stable ampUfication of the nucleic acid sequence can be obtained by integrating at least one additional copy of the sequence into the host ceU genome using methods weU known in the art and selecting for transformants. The procedures used to tigate the elements described above to constmct the recombinant expression vectors of the present invention are weU known to one skilled in the art (see, e.g., Sambrook etal, 1989, supra). Host Cells The present invention also relates to recombinant host ceUs, comprising a nucleic acid sequence of the invention, which are advantageously used in the recombinant production of the first wash Upolytic enzymes. The ceU is preferably transformed with a vector comprising a nucleic acid sequence of the invention foUowed by integration of the vector into the host chromosome. "Transformation" means introducing a vector comprising a nucleic acid sequence of the present invention mto a host ceU so that the vector is maintained as a chromosomal integrant or as a self-repUcating extra-chromosomal vector. Integration is generaUy considered to be an advantage as the nucleic acid sequence is more likely to be stably maintained in the ceU. Integration of the vector into the host chromosome may occur by homologous or non-homologous recombination as described above. The choice of a host ceU will to a large extent depend upon the gene encoding the first wash Upolytic enzyme and its source. In addition, the choice of host ceU will often depend on the proteolytic enzyme system of the host ceU and its impact on the production of a first wash Upolytic enzyme of the invention. Accordingly, it may be desirable to use a host ceU which is deficient in one or more proteolytic enzymes or other enzyme processing means. Protease deficient host ceUs of bacteria as weU as fungal (filamentous fungal and yeast) ceUs are weU-known in the art. When the first wash Upolytic enzyme of the invention comprises a peptide addition it may be advantageous that the host is a strain reduced or deficient in one or more exo-proteases capable of cleaving the modified Upolytic enzyme at a site close to the peptide addition or a protease capable of cleaving within the peptide addition. For instance, the host ceU may be reduced or deficient in a rripeptidyl-aminopeptidase (TPAP) (see e.g. WO 96/14404 from Novo Nordisk A S), a dipeptidyl-aminopeptidase (DPAP), and/or a Kex2 protease or Kex2-like protease and therefore not capable of cleaving at di-basic sites such as Arg-Arg (RR) . Other examples of host ceUs include alkaline protease deficient or reduced host ceUs, aspartic proteinase deficient host cells (EP 429 490), and host ceUs deficient of proteolytic enzymes such as the host ceUs described in WO 93/00925, WO 92/17595, EP 341 215, EP 574 347, and PCT/DK96/00111. The host ceU may be a umceUular microorganism or a non-uniceUular microorganism. Useful uniceUular ceUs are bacterial ceUs such as gram positive bacteria including, but not limited to, a Bacillus ceU, e.g., B. subtilis, B. licheniformis, B. lentus, B. brevis, B. stearothermophilus, B. alkalophilus, B. amyloliquefaciens, B. coagulans, B.circulans, B. lautus, B. megaterium, and B. thuringiensis; or a Streptomyces ceU, e.g., S. lividans or S. murinus, or gram negative bacteria such as E. coli and Pseudomonas sp. (especiaUy when a bacterial Upolytic enzyme, such as a Pseudomonas sp. enzyme is to be produced). The transformation of a bacterial host ceU may, for instance, be effected by protoplast transformation (see, e.g., Chang and Cohen, 1979, Molecular General Genetics 168:111-115), by using competent ceUs (see, e.g., Young and Spizizin, 1961, Journal of Bacteriology 81:823-829, or Dubnar and Davidoff-Abelson, 1971, Journal of Molecular Biology 56:209-221), by electroporation (see, e.g. , Shigekawa and Dower, 1988, Biotechniques 6:742-751), or by conjugation (see, e.g., Koehler and Thome, 1987, Journal of Bacteriology 169:5771-5278). The host ceU may be a eukaryote, and is preferably a fungal, i.e. a yeast ceU or a filamentous fiingal ceU, especiaUy for the production of a modified Upolytic enzyme of eukaryotic origin. "Yeast" as used herein includes ascosporogenous yeast (Endomycetales), basidiosporogenous yeast, and yeast belonging to the Fungi Imperfecti (Blastomycetes). The ascosporogenous yeasts are divided into the families Spermophthoraceae and Saccharomycetaceae. The latter is comprised of four subfamiUes, Schizosaccharomycoideae (e.g., genus Schizosaccharomyces), Nadsonioideae, Lipomycoideae, and Saccharomycoideae (e.g. , genera Pichia, Kluyveromyces and Saccharomyces). The basidiosporogenous yeasts include the genera Leucosporidim, Rhodosporidi m, Sporidiobolus, Fϊlobasidium, and Filobasidiella. Yeast belonging to the Fungi Imperfecti are divided into two families, Sporobolomycetaceae (e.g. , genera Sorobolomyces and Bullera) and Cryptococcaceae (e.g. , genus Candida). Since the classification of yeast may change in the future, for the purposes of this invention, yeast shaU be defined as described in Biology and Activities of Yeast (Skinner, F.A., Passmore, S.M., and Davenport, R.R., eds, Soc. App. Bacteriol. Symposium Series No. 9, 1980. The biology of yeast and manipulation of yeast genetics are weU known in the art (see, e.g. , Biochemistry and Genetics of Yeast, BacU, M. , Horecker, B.J., and Stopani, A.O.M., editors, 2nd edition, 1987; The Yeasts, Rose, A.H., and Harrison, J.S., editors, 2nd edition, 1987; and The Molecular Biology ofthe Yeast Saccharomyces, Strathern et al. , editors, 1981). In connection with the present invention the use of yeast ceUs which typicaUy have another proteolytic enzyme processmg system that, e.g., bacteria and filamentous fungi, may be of particular use for preparing modified Upolytic enzymes which, as the peptide addition, comprise a part or aU of the natural prosequences of the parent Upolytic enzyme in question. When the fungal host ceU is a yeast ceU (e.g. to be used in applying a peptide addition (in the form of part of or the entire prosequence of the parent enzyme, the yeast host ceU may be a ceU of a species of Candida, Kluyveromyces, Saccharomyces, Schizosaccharomyces, Pichia, or Yarrowia, such as & S. cerevisiae ceU, a S.s carlsbergensis, a S. diastaticus ceU, a S. douglasii ceU, a S. kluyveri ceU, a S. norbensis cell, or a S. oviformis ceU. "Fungi" as used herein includes the phyla Ascomycota, Basidiomycota, Chytridiomycota, and Zygomycota (as defined by Hawksworth et al. , In, Ainsworth and Bisby's Dictionary of The Fungi, 8th edition, 1995, CAB Intemational, University Press, Cambridge, UK) as weU as the Oomycota (as cited in Hawksworth et al. , 1995, supra, page 171) and aU mitosporic fiingi (Hawksworth et al, 1995, supra). Representative groups of Ascomycota include, e.g., Neurospora, Eupenicillium (= Penicillium), Emericella (= Aspergillus), Eurotium (= Aspergillus), and the tme yeasts Usted above. Examples of Basidiomycota include mushrooms, rusts, and smuts. Representative groups of Chytridiomycota include, e.g. , Allomyces, Blastocladiella, Coelomomyces, and aquatic fiingi. Representative groups of Oomycota include, e.g., Saprolegniomycetous aquatic fungi (water molds) such as Achlya. Examples of mitosporic fiingi include Aspergillus, Penicillium, Candida, and Alternaria. Representative groups of Zygomycota include, e.g. , Rhizopus and Mucor. "FUamentous fiingi" include aU filamentous forms of the subdivision Eumycota and Oomycota (as defined by Hawksworth et al, 1995, supra). The filamentous fungi are characterized by a vegetative myceUum composed of chitin, ceUulose, glucan, chitosan, mannan, and other complex polysaccharides. Vegetative growth is by hyphal elongation and carbon cataboUsm is obUgately aerobic. In contrast, vegetative growth by yeasts such as Saccharomyces cerevisiae is by budding of a uniceUular thaUus and carbon cataboUsm may be 5 fermentative. In a prefened embodiment, the fungal host ceU is a filamentous fungal ceU. In a more preferred embodiment, the filamentous fungal host ceU is a ceU of a species of, but not Umited to, Acremonium, Aspergillus, Fusarium, Humicola, Myceliophthora, Mucor, Neurospora, Penicillium, Thielavia, Totypocladium, and Trichoderma. In an even more 10 prefened embodiment, the filamentous fungal host ceU is an Aspergillus cell. In another even more preferred embodiment, the filamentous fungal host ceU is a Fusarium ceU. In a most prefened embodiment, the filamentous fungal host ceU is an A. oryzae ceU, an A. niger ceU, an A. foetidus ceU, or an A. japonicus ceU. In another most prefened embodiment, the filamentous fungal host ceU is a Fusarium oxysporum ceU or a F. graminearum ceU. 15 Fungal ceUs may be transformed by a process involving protoplast formation, transformation of the protoplasts, and regeneration of the ceU waU in a manner known er -ye. Suitable procedures for transformation of Aspergillus host ceUs are described in EP 238 023 and Yelton et al., 1984, Proceedings ofthe National Academy of Sciences USA 81:1470- 1474. A suitable method of transforming Fusarium species is described by Malardier et al. , 20 1989, Gene 78: 147-156 or in WO 96/00787. Yeast may be transformed using the procedures described by Becker and Guarente, In Abelson, J.N. and Simon, M.I., editors, Guide to Yeast Genetics and Molecular Biology, Methods in Enzymology, Volume 194, pp 182-187, Academic Press, Inc., New York; Ito et al., 1983, Journal of Bacteriology 153:163; and Hinnen et al, 1978, Proceedings ofthe National Academy of Sciences USA 75:1920. 25 MammaUan ceUs may be transformed by direct uptake using the calcium phosphate precipitation method of Graham and Van der Eb (1978, Virology 52:546). Methods of Production The present invention also relates to methods for producing a first wash Upolytic 30 enzyme of the invention comprismg (a) cultivating a host ceU transformed with a DNA sequence encoding the enzyme under conditions conducive to expression of the first wash Upolytic enzyme; and (b) recovering the first wash Upolytic enzyme. The host ceUs may be cultivated in a nutrient medium suitable for production of the first wash Upolytic enzyme using methods known in the art. For example, the cell may be cultivated by shake flask cultivation, smaU-scale or large-scale fermentation (including continuous, batch, fed-batch, or sotid state fermentations) in laboratory or industrial fermentors performed in a suitable medium and under conditions aUowing the modified Upolytic enzyme to be expressed and/or isolated. The cultivation takes place in a suitable nutrient medium comprising carbon and nitrogen sources and inorganic salts, using procedures known in the art (see, e.g. , references for bacteria and yeast; Bennett, J.W. and LaSure, L., editors, More Gene Manipulations in Fungi, Academic Press, CA, 1991). Suitable media are available from commercial suppUers or may be prepared accordmg to pubUshed compositions (e.g. , in catalogues of the American Type Culture CoUection). If the first wash Upolytic enzyme is secreted into the nutrient medium, the Upolytic enzyme can be recovered directly from the medium. If the first wash Upolytic enzyme is not secreted, it is recovered from ceU lysates. The resulting first wash Upolytic enzyme may be recovered by methods known in the art. For example, the first wash Upolytic enzyme may be recovered from the nutrient medium by conventional procedures including, but not timited to, centrifugation, filtration, extraction, spray-drying, evaporation, or precipitation. The recovered first wash Upolytic enzyme may then be further purified by a variety of chromatographic procedures, e.g. , ion exchange chromatography, gel filtration chromatography, affinity chromatography, or the Uke. The first wash Upolytic enzymes of the present invention may be purified by a variety of procedures known in the art including, but not timited to, chromatography (e.g. , ion exchange, affinity, hydrophobic, chromatofocusing, and size exclusion), electrophoretic procedures (e.g., preparative isoelectric focusing (TEF), differential solubiUty (e.g., ammomum sulfate precipitation), or extraction (see, e.g. , Protein Purification, J.-C. Janson and Lars Ryden, editors, VCH PubUshers, New York, 1989). In accordance with the invention, it is also contemplated to apply, to the first wash Upolytic Upolytic enzyme, one or more charged amino acids which permit effective purifica¬ tion of the modified enzyme. Techniques for doing this is weU known by a person skilled in the art of molecular biology. The host ceU, cultivation condition and/or recovery conditions are preferably selected so that at the most a partial processing of the pre, pro or preproform of the parent enzyme has occured resulting in that at least 5 % , such as at least 10% , such as at least 15 % , such as at least 20% , such as at least 25% , such as at least 50% or at least 75% of the produced modi¬ fied enzyme molecules comprises the desired , e.g. the entire pre-sequence or a substantial part thereof. Enzyme composition of the invention In a further aspect the invention relates to an enzyme composition comprising a first wash Upolytic enzyme of the invention. As defined herein, a "substantiaUy pure" enzyme is an enzyme which is essentiaUy free of other homologous contaminants (originating from the same source as the first wash Upolytic enzyme), e.g. , at least about 20% pure, preferably at least about 40% pure, more preferably about 60% pure, even more preferably about 80% pure, most preferably about 90% pure, and even most preferably about 95% pure, as determined by SDS-PAGE. When the first wash Upolytic enzyme of the invention comprises a peptide addition it has been found that not aU of the first wash Upolytic enzyme molecules expressed by a given host cell are processed at the same cleavage site. This has the consequence that the first wash Upolytic enzyme product recovered from the fermentation by such host ceUs comprise a por¬ tion having the fuU length peptide addition and one or more other portions with only a part of the peptide addition. The inventors found that this does not influence the wash performance significantly. Consequently, even though not aU of the Upolytic enzyme of the enzyme com¬ position of the invention may have retained the full length peptide addition the enzyme com¬ position is still capable of exerting a first wash effect. ActuaUy, it has been found that as long as at least about 5% of the total amount of first wash Upolytic enzyme of the invention con¬ taining a peptide addition has the intact peptide addition as disclosed above, this may be found to be sufficient for providing the desired first wash effect. The remaining part of the first wash Upolytic enzyme molecules may then have a peptide addition which is shorter than the one intended (e.g. as a consequence of one or more amino acid residues have been cut off during processing of the enzyme by the host organism). Therefore, the enzyme composition of the mvention need only to comprise at least about 5%, preferably at least about 10%, such as at least about 25 % , better at least about 50 % , especiaUy at least about 75 % of the first wash Upolytic enzyme with its fuU length addition. Said enzyme composition may further comprise an enzyme selected from the group of proteases, ceUulases, peroxidases, cutinases, amylases and or Upases, and when intended for washing also ingredients normaUy used in detergent compositions. DETERGENT DISCLOSURE Surfactant system • The detergent compositions according to the present invention comprise a surfactant system wherein the surfactants can be selected from nonionic and/or anionic and/or catio¬ nic and/or ampholytic and/or zwitterionic and/or semi-polar surfactants. The surfactants are typically present at a level of from 0.1 % to 60% by weight. The surfactant is preferably formulated to be compatible with enzyme components present in the composition. In liquid or gel compositions the surfactant is most preferably formulated such that it promotes, or at least does not degrade, the stability of any enzyme in these compositions. Prefened surfactant systems to be used according to the present invention comprise as a surfactant one or more of the nonionic and/or anionic surfactants described herein. Nonionic detergent surfactants normally consist of a water-solubilizing polyalkoxy¬ lene or a mono- or di-alkanolamide group in chemical combination with an organic hy¬ drophobic group derived, for example, from alkylphenols in which the alkyl group con¬ tains from about 6 to about 12 carbon atoms, dialkylphenols in which each alkyl group contains from 6 to 12 carbon atoms, primary, secondary or tertiary aliphatic alcohols (or alkyl-capped derivatives thereof), preferably having from 8 to 20 carbon atoms, mono¬ carboxylic acids having from 10 to about 24 carbon atoms in the alkyl group and polyoxypropylenes. Also common are fatty acid mono- and dialkanolamides in which the alkyl group of the fatty acid radical contains from 10 to about 20 carbon atoms and the alkyloyl group having from 1 to 3 carbon atoms. Optionally, in any of the mono- and di- alkanolamide derivatives there may be a polyoxyalkylene moiety joining the latter groups and the hydrophobic part of the molecule. In all polyalkoxylene containing surfactants, the polyalkoxylene moiety preferably consists of from 2 to 20 groups of ethylene oxide or of ethylene oxide and propylene oxide groups. Polyethylene, polypropylene, and polybutylene oxide condensates of alkyl phenols are suitable for use as the nonionic surfactant of the surfactant systems of the present in¬ vention, with the polyethylene oxide condensates being preferred. These compounds in¬ clude the condensation products of alkyl phenols having an alkyl group containing from about 6 to about 14 carbon atoms, preferably form about 8 to about 14 carbon atoms, in either a straightchain or branched-chain configuration with the alkylene oxide. In a pre¬ ferred embodiment, the ethylene oxide is present in an amount equal to from about 2 to about 25 moles, more preferably from about 3 to about 15 moles, of ethylene oxide per mole of alkyl phenol. Commercially available nonionic surfactants of this type include Igepal™ CO-630, marketed by the GAF Coφoration; and Triton™ X-45, X-114, X-100 and X-102, aU marketed by the Rohm & Haas Company. These surfactants are com¬ monly referred to as alkylphenol alkoxylates (e.g., alkyl phenol ethoxylates). The condensation products of primary and secondary aliphatic alcohols with from about 1 to about 25 moles of ethylene oxide are suitable for use as the nonionic surfactant of the nonionic surfactant systems of the present invention. The alkyl chain of the alipha¬ tic alcohol can either be straight or branched, primary or secondary, and generally con¬ tains from about 8 to about 22 carbon atoms. Preferred are the condensation products of alcohols having an alkyl group containing from about 8 to about 20 carbon atoms, more preferably from about 10 to about 18 carbon atoms, with from about 2 to about 10 moles of ethylene oxide per mole of alcohol. About 2 to about 7 moles of ethylene oxide and most preferably from 2 to 5 moles of ethylene oxide per mole of alcohol are present in said condensation products. Examples of commercially available nonionic surfactants of this type include Tergitol™ 15-S-9 (The condensation product of C n -C ι5 linear alcohol with 9 moles ethylene oxide), Tergitol 24-L-6 NMW (the condensation product of C n C M primary alcohol with 6 moles ethylene oxide with a narrow molecular weight distri¬ bution), both marketed by Union Carbide Coφoration; Neodol™ 45-9 (the condensation product of C ]4 -C, 5 linear alcohol with 9 moles of ethylene oxide), Neodol™ 23-3 (the condensation product of C, 2 -C 13 linear alcohol with 3.0 moles of ethylene oxide), Neo " dol™ 45-7 (the condensation product of Cj 4 -C, 5 tinear alcohol with 7 moles of ethylene oxide), Neodol™ 45-5 (the condensation product of C 14 -C 15 linear alcohol with 5 moles of ethylene oxide) marketed by Shell Chemical Company, Kyro™ EOB (the condensation product of C 13 -Ci 5 alcohol with 9 moles ethylene oxide), marketed by The Procter & Gamble Company, and Genapol LA 050 (the condensation product of C ι2 -C 14 alcohol with 5 moles of ethylene oxide) marketed by Hoechst. Preferred range of HLB in these products is from 8-11 and most preferred from 8-10. The detergent composition of the invention may comprise a nonionic material which is alkoxylated aliphatic alcohols containing at least 25 alkylene oxide groups, preferably at least 50 alkylene oxide groups, and more preferably at least 80 alkylene oxide groups. Also useful as the nonionic surfactant of the surfactant systems of the present in¬ vention are alkylpolysaccharides disclosed in US 4,565,647, having a hydrophobic group containing from about 6 to about 30 carbon atoms, preferably from about 10 to about 16 carbon atoms and a polysaccharide, e.g. a polyglycoside, hydrophilUc group containing from about 1.3 to about 10, preferably from about 1.3 to about 3, most preferably from about 1.3 to about 2.7 saccharide units. Any reducing saccharide containing 5 or 6 car¬ bon atoms can be used, e.g., glucose, galactose and galactosyl moieties can be substituted for the glucosyl moieties (optionally the hydrophobic group is attached at the 2-, 3-, 4-, etc. positions thus giving a glucose or galactose as opposed to a glucoside or galactoside). The intersaccharide bonds can be, e.g., between the one position of the additional saccha¬ ride units and the 2-, 3-, 4-, and/or 6- positions on the preceding saccharide units. Another useful nonionic surfactant is a glycoside of a uronic acid, a uronic acid salt or a uronic acid lactone or polyuronic acid with a straight-chain or branced saturated or unsaturated aliphatic chain of from 6 to 24 carbon atoms, optionally containing an aro¬ matic, cycloaliphatic, mixed aromatic-aliphatic or polyalkyloxyalkyl radical, as described in WO 95/10524. The preferred alkylpolyglycosides have the formula R 2 O(C n H 2n O) l (glycosyl) x wherein R 2 is selected from the group consisting of alkyl, alkylphenyl, hydroxyalkyl, hy¬ droxyalkylphenyl, and mixtures thereof in which the alkyl groups contain from about 10 to about 18, preferably from about 12 to about 14, carbon atoms; n is 2 or 3, preferably 2; t is from 0 to about 10, preferably 0; and x is from about 1.3 to about 10, preferably from about 1.3 to about 3, most preferably from about 1.3 to about 2.7. The glycosyl is preferably derived from glucose. To prepare these compounds, the alcohol or alkylpoly- ethoxy alcohol is formed first and then reacted with glucose, or a source of glucose, to form the glucoside (attachment at the 1-position). The additional glycosyl units can then be attached between their 1 -position and the preceding glycosyl units 2-, 3-, 4-, and/or 6- position, preferably predominately the 2-position. The condensation products of ethylene oxide with a hydrophobic base formed by the condensation of propylene oxide with propylene glycol are also suitable for use as the additional nonionic surfactant systems of the present invention. The hydrophobic portion of these compounds will preferably have a molecular weight of from about 1500 to about 1800 and will exhibit water insolubiUty. The addition of polyoxyethylene moieties to this hydrophobic portion tends to increase the water solubiUty of the molecule as a whole, and the liquid character of the product is retained up to the point where the polyoxyethylene content is about 50% of the total weight of the condensation product, which corresponds to condensation with up to about 40 moles of ethylene oxide. Examples of compounds of this type include certain of the commercially avaUable Pluronic™ surfactants, marketed by BASF. Also suitable for use as the nonionic surfactant of the nonionic surfactant system of the present invention, are the condensation products of ethylene oxide with the product resulting from the reaction of propylene oxide and ethylenediamine. The hydrophobic moiety of these products consists of the reaction product of ethylenediamine and excess propylene oxide, and generally has a molecular weight of from about 2500 to about 3000. This hydrophobic moiety is condensed with ethylene oxide to the extent that the conden¬ sation product contains from about 40% to about 80% by weight of polyoxyethylene and has a molecular weight of from about 5,000 to about 11.000. Examples of this type of nonionic surfactant include certain of the commerciaUy available Tetronic™ compounds, marketed by BASF. Preferred for use as the nonionic surfactant of the surfactant systems of the present invention are polyethylene oxide condensates of alkyl phenols, condensation products of primary and secondary aliphatic alcohols with from about 1 to about 25 moles of ethyle- neoxide, alkylpoly saccharides, and mixtures hereof. Most preferred are C 8 -C ι4 alkyl phe- nol ethoxylates having from 3 to 15 ethoxy groups and C 8 -C 18 alcohol ethoxylates (preferably C^ avg.) having from 2 to 10 ethoxy groups, and mixtures thereof. Highly preferred nonionic surfactants are polyhydroxy fatty acid amide surfactants of the formula R 2 - C - N - Z, 0 R 1 wherein R 1 is H, or R 1 is C M hydrocarbyl, 2-hydroxy ethyl, 2-hydroxy propyl or a mix- ture thereof, R 2 is C 5 . 31 hydrocarbyl, and Z is a polyhydroxyhydrocarbyl having a Unear hydrocarbyl chain with at least 3 hydroxyls directly connected to the chain, or an al¬ koxylated derivative thereof. Preferably, R 1 is methyl, R 2 is straight C 1 5 alkyl or C 16 . 18 alkyl or alkenyl chain such as coconut alkyl or mixtures thereof, and Z is derived from a reducing sugar such as glucose, fructose, maltose, lactose, in a reductive amination reac- tion. A nonionic surfactant system comprising an alkoxylated nonionic surfactant having an average degree of alkoxylation of at least 6 and an aldobionamide of the stmcture ANRιR 2 wherein A is a sugar moiety which is an aldobionic acid except that it does not contain the OH group normally extending from the carbonyl group on the aldonic acid. NRjR 2 is attached where the hydroxyl group on the aldobionic acid would normally be found. RιR 2 , may be the same or different, is a hydrogen atom, an aUphatic hydrocarbon radical, an aromatic radical a cycloaliphatic radical an amino acid ester, or an ether ami¬ ne. Ri and R 2 cannot both be hydrogen atoms as described in WO 95/2770. Other so-called nonionic detergent compounds include long chain tertiary amine oxides, long chain tertiary phosphine oxides and dialkyl sulphoxides. Highly prefened anionic surfactants include alkyl alkoxylated sulfate surfactants hereof are water soluble salts or acids of the formula RO(A) m SO 3 M wherein R is an un¬ substituted C 10 -C- 24 alkyl or hydroxyalkyl group having a C 10 -C 24 alkyl component, prefe- rably a Cι 2 -C 20 alkyl or hydroxyalkyl, more preferably C 12 -C, 8 alkyl or hydroxyalkyl, A is an ethoxy or propoxy unit, m is greater than zero, typically between about 0.5 and about 6, more preferably between about 0.5 and about 3, and M is H or a cation which can be, for example, a metal cation (e.g., sodium, potassium, lithium, calcium, magnesi¬ um, etc.), ammonium or substituted-ammonium cation. Alkyl ethoxylated sulfates as well as alkyl propoxylated sulfates are contemplated herein. Specific examples of substituted ammonium cations include methyl-, dimethyl, trimethyl-ammonium cations and quatema¬ ry ammonium cations such as tetramethyl-ammonium and dimethyl piperdinium cations and those derived from alkylamines such as ethylamine, diethylamine, triethylamine, mixtures thereof, and the like. Exemplary surfactants are C 12 -Cι 8 alkyl polyethoxylate (1.0) sulfate (C 12 -Cι 8 E(1.0)M), C 12 -C 18 alkyl polyethoxylate (2.25) sulfate (C 12 - C 18 (2.25)M, and C, 2 -C 18 alkyl polyethoxylate (3.0) sulfate (C, 2 -Cι 8 E(3.0)M), and C 12 -C 18 alkyl polyethoxylate (4.0) sulfate (Cι 2 -C 18 E(4.0)M), wherein M is conveniently selected from sodium and potassium. Suitable anionic surfactants to be used are alkyl ester sul¬ fonate surfactants including linear esters of C 8 -C 20 carboxylic acids (i.e., fatty acids) which are sulfonated with gaseous SO 3 according to "The Joumal of the American Oil Chemists Society" ' , 52 (1975), pp. 323-329. Suitable starting materials would include natural fatty substances as derived from tallow, palm oil, etc. The preferred alkyl ester sulfonate surfactant, especially for laundry applications, comprise alkyl ester sulfonate surfactants of the stmctural formula: 0 R 3 - CH - C - OR 4 SO,M wherein R 3 is a C 8 -C 20 hydrocarbyl, preferably an alkyl, or combination thereof, R 4 is a Cι-C 6 hydrocarbyl, preferably an alkyl, or combination thereof, and M is a cation which forms a water soluble salt with the alkyl ester sulfonate. Suitable salt-forming cations in¬ clude metals such as sodium, potassium, and lithium, and substituted or unsubstituted ammomum cations, such as monoethanolamine, diethonolamine, and triethanolamine. Preferably, R 3 is C 10 -C, 6 alkyl, and R 4 is methyl, ethyl or isopropyl. Especially preferred are the methyl ester sulfonates wherein R 3 is C, 0 -C 16 alkyl. Other suitable anionic surfactants include the alkyl sulfate surfactants which are water soluble salts or acids of the formula ROSO 3 M wherein R preferably is a Cι 0 -C 24 hydrocarbyl, preferably an alkyl or hydroxyalkyl having a C, 0 -C 2 o alkyl component, more preferably a Cι 2 -Cι 8 alkyl or hydroxyalkyl, and M is H or a cation, e.g., an alkati metal cation (e.g. sodium, potassium, lithium), or ammonium or substituted ammonium (e.g. methyl-, dimethyl-, and trimethyl ammonium cations and quatemary ammonium cations such as tetramethyl-ammonium and dimethyl piperdinium cations and quatemary ammo¬ nium cations derived from alkylamines such as ethylamine, diethylamine, triethylamine, and mixtures thereof, and the Uke). Typically, alkyl chains of C 12 -Cι 6 are preferred for lower wash temperatures (e.g. below about 50°C) and C 16 -C 18 alkyl chains are preferred for higher wash temperatures (e.g. above about 50°C). Other anionic surfactants useful for detersive puφoses can also be included in the laundry detergent compositions of the present invention. Theses can include salts (including, for example, sodium, potassium, ammonium, and substituted ammomum salts such as mono- di- and triethanolamine salts) of soap, C 8 -C 22 primary or secondary alkane- sulfonates, C 8 -C 24 olefinsulfonates, sulfonated polycarboxylic acids prepared by sulfonati¬ on of the pyrolyzed product of alkaline earth metal citrates, e.g., as described in GB 1,082,179, C 8 -C 24 alkylpolyglycolethersulfates (containing up to 10 moles of ethylene oxide); alkyl glycerol sulfonates, fatty acyl glycerol sulfonates, fatty oleyl glycerol sulfa¬ tes, alkyl phenol ethylene oxide ether sulfates, paraffin sulfonates, alkyl phosphates, iset- hionates such as the acyl isethionates, isethionates esters of alkoxy carboxylic acids (as described in WO95/14661), N-acyl taurates, alkyl succinamates and sulfosuccinates, mo¬ noesters of sulfosuccinates (especially saturated and unsaturated C 12 -Cι 8 monoesters) and diesters of sulfosuccinates (especially saturated and unsaturated C 6 -C 12 diesters), acyl sar¬ cosinates, oleoyl sarcosinate, sulfates of alkylpolysaccharides such as the sulfates of al- kylpolyglucoside (the nonionic nonsulfated compounds being described below), branched primary alkyl sulfates, and alkyl polyethoxy carboxylates such as those of the formula RO(CH 2 CH 2 O) k -CH 2 C00-M+ wherein R is a C 8 -C 22 alkyl, k is an integer from 1 to 10, and M is a soluble salt forming cation. Resin acids and hydrogenated resin acids are also suitable, such as rosin, hydrogenated rosin, and resm acids and hydrogenated resin acids present in or derived from tall oil. Alkylbenzene sulfonate is highly preferred, in parti¬ cular linear alkyl benzene sulfonate (LAS) where the alkyl group contains preferably from 10 to 18 carbon atoms. Further examples are described in "Surface Active Agents and Detergents" (Vol. I and II by Schwartz, Perrry and Berch). A variety of such surfactants are also generally disclosed in US 3,929,678, (Column 23, line 58 through Column 29, line 23). When mcluded therein, the laundry detergent compositions of the present invention typically comprise from about 1% to about 40%, preferably from about 3% to about 20% by weight of such anionic surfactants. The laundry detergent compositions of the present invention may also contain catio- nic, ampholytic, zwitterionic, and semi-polar surfactants, as well as the nonionic and/or anionic surfactants other than those already described herein. Cationic detersive surfactants suitable for use in the laundry detergent compositions of the present invention are those having one long-chain hydrocarbyl group. Examples of such cationic surfactants include the ammonium surfactants such as alkyltrimethylammo- nium halogenides, and those surfactants having the formula: [R 2 (OR 3 ) y ][R 4 (OR 3 ) y ] 2 R s N-l-X- wherein R 2 is an alkyl or alkyl benzyl group having from about 8 to about 18 carbon atoms in the alkyl chain, each R 3 is selected form the group consisting of -CH 2 CH 2 -, - CH 2 CH(CH 3 )-, -CH 2 CH(CH 2 OH)-, -CH 2 CH 2 CH 2 -, and mixtures thereof; each R 4 is se¬ lected from the group consisting of C r C 4 alkyl, - hydroxyalkyl, benzyl ring stmctu¬ res formed by joining the two R 4 groups, -CH 2 CHOHCHOHCOR 6 CHOHCH 2 OH whe¬ rein R 6 is any hexose or hexose polymer having a molecular weight less than about 1000, and hydrogen when y is not 0; R 5 is the same as R 4 or is an alkyl chain wherein the total number of carbon atoms or R 2 plus R 5 is not more than about 18; each y is from 0 to about 10 and the sum of the y values is form 0 to about 15; and X is any compatible ani¬ on. Highly preferred cationic surfactants are the water soluble quatemary ammonium compounds useful in the present composition having the formula: R 1 -R 2 R 9 R 4 N + X ' (i) wherein R ! is C 8 -Cι 6 alkyl, each of R 2 , R 3 and R, is independently C,-C 4 alkyl, C r C 4 hydroxy alkyl, benzyl, and -(C 2 H 40 ) X H where x has a value from 2 to 5, and X is an ani¬ on. Not more than one of R 2 , R 3 or R, should be benzyl. The preferred alkyl chain length for Rj is Cι 2 -Cι 5 particularly where the alkyl group is a mixture of chain lengths derived from coconut or palm kernel fat or is derived syn¬ thetically by olefin build up or OXO alcohols synthesis. Preferred groups for R 2 R 3 and R, are methyl and hydroxyethyl groups and the anion X may be selected from halide, methosulphate, acetate and phosphate ions. Exam¬ ples of suitable quatemary ammonium compounds of formulae (i) for use herein are: coconut trimethyl ammonium chloride or bromide; coconut methyl dihydroxyethyl ammonium chloride or bromide; decyl triethyl ammonium chloride; decyl dimethyl hydroxyethyl ammonium chloride or bromide; C 12 .ιs dimethyl hydroxyethyl ammonium chloride or bromide; coconut dimethyl hydroxyethyl ammonium chloride or bromide; myristyl trimethyl ammonium methyl sulphate; lauryl dimethyl benzyl ammonium chloride or bromide; lauryl dimethyl (ethenoxy) 4 ammonium chloride or bromide; choline esters (compounds of formula (i) wherein R, is CH 2 -CH 2 -O-C-C 12 . 14 alkyl and R 2 R 3 R-, are methyl). II O di-alkyl imidazolines [compounds of formula (i)]. Other cationic surfactants useful herein are also described in US 4,228,044 and in EP 000 224. When included therein, the laundry detergent compositions of the present invention typically comprise from 0.2% to about 25%, preferably from about 1 % to about 8% by weight of such cationic surfactants. Ampholytic surfactants are also suitable for use in the laundry detergent compositi¬ ons of the present invention. These surfactants can be broadly described as aliphatic deri¬ vatives of secondary or tertiary amines, or ahphatic derivatives of heterocyclic secondary and tertiary amines in which the aUphatic radical can be straight- or branched-chain. One of the aUphatic substituents contains at least about 8 carbon atoms, typically from about 8 to about 18 carbon atoms, and at least one contains an anionic water-solubiUzing group, e.g. carboxy, sulfonate, sulfate. See US 3,929,678 (column 19, Unes 18-35) for exam- pies of ampholytic surfactants. When included therein, the laundry detergent compositions of the present invention typically comprise from 0.2% to about 15%, preferably from about 1% to about 10% by weight of such ampholytic surfactants. Zwitterionic surfactants are also suitable for use in laundry detergent compositions. These surfactants can be broadly described as derivatives of secondary and tertiary ami¬ nes, derivatives of heterocyclic secondary and tertiary amines, or derivatives of quatema¬ ry ammonium, quatemary phosphonium or tertiary sulfonium compounds. See US 3,929,678 (column 19, line 38 through column 22, Une 48) for examples of zwitterionic surfactants. When included therein, the laundry detergent compositions of the present invention typically comprise from 0.2% to about 15%, preferably from about 1 % to about 10% by weight of such zwitterionic surfactants. Semi-polar nonionic surfactants are a special category of nonionic surfactants which in- elude water-soluble amine oxides containing one alkyl moiety of from about 10 to about 18 carbon atoms and 2 moieties selected from the group consisting of alkyl groups and hydroxyalkyl groups containing from about 1 to about 3 carbon atoms; watersoluble phosphine oxides containing one alkyl moiety of form about 10 to about 18 carbon atoms and 2 moieties selected from the group consisting of alkyl groups and hydroxyalkyl groups containing from about 1 to about 3 carbon atoms; and water-soluble sulfoxides containing one alkyl moiety of from about 10 to about 18 carbon atoms and a moiety se¬ lected from the group consisting of alkyl and hydroxyalkyl moieties of from about 1 to about 3 carbon atoms. Semi-polar nonionic detergent surfactants include the amine oxide surfactants ha- ving the formula: O I R 3 (OR 4 )xN(R 5 )2 wherein R 3 is an alkyl, hydroxyalkyl, or alkyl phenyl group or mixtures thereof contai- ning from about 8 to about 22 carbon atoms; R 4 is an alkylene or hydroxyalkylene group containing from about 2 to about 3 carbon atoms or mixtures thereof; x is from 0 to about 3: and each R 5 is an alkyl or hydroxyalkyl group containing from about 1 to about 3 car¬ bon atoms or a polyethylene oxide group containing from about 1 to about 3 ethylene oxide groups. The'R 5 groups can be attached to each other, e.g., through an oxygen or nitrogen atom, to form a ring structure. These amine oxide surfactants in particular include C 10 -C I8 alkyl dimethyl amine oxides and C 8 -C 12 alkoxy ethyl dihydroxy ethyl amine oxides. When included therein, the laundry detergent compositions of the present invention typically comprise from 0.2% to about 15%, preferably from about 1 % to about 10% by weight of such semi-polar nonionic surfactants. Builder system The compositions according to the present invention may further comprise a builder system. Any conventional builder system is suitable for use herein including aluminosiU- cate materials, sUicates, polycarboxylates and fatty acids, materials such as ethylenedia¬ mine tetraacetate, metal ion sequestrants such as aminopolyphosphonates, particularly ethylenediamine tetramethylene phosphonic acid and diethylene triamine pentamethyle¬ nephosphonic acid. Phosphate buUders can also be used herein, e.g pyrophosphates, ort¬ hophosphates or polyphosphates. Suitable buUders can be an inorganic ion exchange material, commonly an inorga¬ nic hydrated aluminosilicate material, more particularly a hydrated synthetic zeolite such as hydrated zeolite A, X, B, HS or MAP. Another suitable inorganic builder material is layered silicate, e.g. SKS-6 (Hoechst). SKS-6 is a crystalline layered sUicate consisting of sodium siUcate (Na 2 Si 2 O 5 ). Suitable polycarboxylates containing one carboxy group include lactic acid, glycolic acid and ether derivatives thereof as disclosed in BE 831,368, BE 821,369 and BE 821,370. Polycarboxylates containing two carboxy groups include the water-soluble salts of succinic acid, malonic acid, (ethylenedioxy) diacetic acid, maleic acid, diglycoltic a- cid, tartaric acid, tartronic acid and fumaric acid, as well as the ether carboxylates de¬ scribed in DE 2,446,686, DE 2,446,487, US 3,935,257 and the sulfinyl carboxylates de¬ scribed in BE 840,623. Polycarboxylates containing three carboxy groups include, in particular, water-soluble citrates, aconitrates and citraconates as weU as succinate deriva¬ tives such as the carboxymethyloxysuccinates described in GB 1,379,241, lac- toxysuccinates described in NL application 7205873, and the oxypolycarboxylate materi¬ als such as 2-oxa-l,l,3-propane tricarboxylates described in GB 1,387,447. Polycarboxylates containing four carboxy groups include oxydisuccinates disclosed in GB 1,261,829, 1,1,2,2,-ethane tetracarboxylates, 1,1,3,3-propane tetracarboxylates containing sulfo substituents include the sulfosuccinate derivatives disclosed in GB 1,398,421 and GB 1,398,422 and in US 3,936,448, and the sulfonated pyrolysed citrates described in GB 1,082,179, while polycarboxylates containing phosphone substituents are disclosed in GB 1,439,000. Alicyclic and heterocyclic polycarboxylates include cis,cis-cis-cyclopentane- tetracarboxylates, pentacarboxylatocyclopentadiene, cis, cis, cis-tetrahydrofuran-tetra- carboxylates, cis-tetrahydrofuran-2.5-dicarboxylates. tetrahydrofuran-2,2,5,5,- tetracarboxylates, hexane-1, 2,3,4,5, 6-hexacarboxylates and carboxymethyl derivatives of polyhydric alcohols such as sorbitol, mannitol and xylitol. Aromatic polycarboxylates in- elude meUitic acid, pyromellitic acid and the phthalic acid derivatives disclosed in GB 1,425,343. Of the above, the preferred polycarboxylates are hydroxycarboxylates contai¬ ning up to three carboxy groups per molecule, more particularly citrates. Preferred builder systems for use in the present compositions include a mixture of a water-insoluble aluminosilicate builder such as zeotite A or of a layered silicate (SKS-6), and a water-soluble carboxylate chelating agent such as citric acid. A suitable chelant for inclusion in the detergent compositions in accordance with the invention is ethylenediamine-N,N'-disuccinic acid (EDDS) or the alkaU metal, alkaline earth metal, ammonium, or substituted ammonium salts thereof, or mixtures thereof. Pre¬ ferred EDDS compounds are the free acid form and the sodium or magnesium salt there- of. Examples of such preferred sodium salts of EDDS include Na 2 EDDS and Na^DDS. Examples of such preferred magnesium salts of EDDS include MgEDDS and Mg 2 EDDS. The magnesium salts are the most preferred for inclusion in compositions in accordance with the invention. Preferred buUder systems include a mixture of a water-insoluble aluminosiUcate builder such as zeolite A, and a water soluble carboxylate chelating agent such as citric acid. Other builder materials that can form part of the builder system for use in granular compositions include inorganic materials such as alkali metal carbonates, bicarbonates, si- ticates, and organic materials such as the organic phosphonates, amino polyalkylene phosphonates and amino polycarboxylates. Other suitable water-soluble organic salts are the homo- or co-polymeric acids or their salts, in which the polycarboxylic acid comprises at least two carboxyl radicals sepa¬ rated form each other by not more than two carbon atoms. Polymers of this type are disclosed in GB-A-1,596,756. Examples of such salts are polyacrylates of MW 2000-5000 and their copolymers with maleic anhydride, such co- polymers having a molecular weight of from 20,000 to 70,000, especially about 40,000. Detergency builder salts are normally included in amounts of from 5% to 80% by weight of the composition. Preferred levels of builder for liquid detergents are from 5% to 30%. Enzymes Preferred detergent compositions, in addition to the enzyme of the invention, co¬ mprise other enzyme(s) which provides cleaning performance and/or fabric care benefits. Such enzymes include proteases, lipases, cutinases, amylases, cellulases, peroxidases, oxidases (e.g. laccases). Proteases: Any protease suitable for use in alkaline solutions can be used. Suitable pro¬ teases include those of animal, vegetable or microbial origin. Microbial origin is pre¬ ferred. Chemically or genetically modified mutants are included. The protease may be a serine protease, preferably an alkaline microbial protease or a trypsin-like protease. E- xamples of alkaline proteases are subtiUsins, especially those derived from Bacillus, e.g., subtilisin Novo, subtilisin Carlsberg, subtiUsin 309, subtilisin 147 and subtiUsin 168 (described in WO 89/06279). Examples of trypsin-like proteases are trypsin (e.g. of por¬ cine or bovine origin) and the Fusarium protease described in WO 89/06270. Prefened commercially avaUable protease enzymes include those sold under the tradenames Alcala- se, Savinase, Primase, Durazym, and Esperase by Novo Nordisk A/S (Denmark), those sold under the tradename Maxatase, Maxacal, Maxapem and Properase by Gist-Bro- cades/Genencor, those sold under the tradename Purafect and Purafect OXP by Genencor Intemational, and those sold under the tradename Opticlean and Optimase by Solvay En¬ zymes. Protease enzymes may be incoφorated into the compositions in accordance with the invention at a level of from 0.0001 % to 2% of enzyme protein by weight of the com¬ position, in particular at a level of from 0.001 % to 0.1 % of enzyme protein by weight of the composition. Lipases: Any lipase suitable for use in alkaline solutions can be used. Suitable lipases in¬ clude those of bacterial or fungal origin and chemically or genetically modified Upase mutants. Examples of useful lipases include a Humicola lanuginosa lipase, e.g., as described in EP 258 068 and EP 305 216, and mutants thereof as described in WO 92/05249, WO 94/25577 and WO 95/22615 a Rhizomucor miehei lipase, e.g., as described in EP 238 023, a Candida lipase, such as a C. antarctica lipase, e.g., the C. antarctica lipase A or B described in EP 214 761, a Pseudomonas lipase such as a P. alcaligenes and P. pseudoal¬ caligenes lipase, e.g., as described in EP 218 272, or any mutant of said Pseudomonas lipases, a P. cepacia lipase, e.g., as described in EP 331 376, a P. stutzeri Upase, e.g., as disclosed in BP 1,372,034, a P. fluorescens lipase, a Bacillus lipase. e.g., a B. subtilis lipase (Dartois et al., (1993), Biochemica et Biophysica acta 1131, 253-260), a B. stearo¬ thermophilus lipase (JP 64/744992) and a B. pumilus lipase (WO 91/16422). Furthermo¬ re, a number of cloned lipases may be useful, including the Penicillium camembertii lipa¬ se described by Yamaguchi et al., (1991), Gene 103, 61-67), the Geotricum candidum lipase (Schimada, Y. et al., (1989), J. Biochem., 106, 383-388), and various Rhizopus lipases such as a R. delemar Upase (Hass, MJ et al., (1991), Gene 109, 117-113), a J niveus lipase (Kugimiya et al., (1992), Biosci. Biotech. Biochem. 56, 716-719) and a R. oryzae lipase. Other types of lipolytic enzymes such as cutinases may also be useful, e.g. , a cuti- nase derived from Pseudomonas mendocina as described in WO 88/09367, or a cutinase derived from Fusarium solani pisi (e.g. described in WO 90/09446). Especially suitable lipases are lipases such as Ml Lipase™, Luma fast™ and Lipomax™ (Gist-Broca- des/Genencor), Lipolase™ and Lipolase Ultra™ (Novo Nordisk A/S), and Lipase P "Amano" (Amano Pharmaceutical Co. Ltd.). The Upases are normally incoφorated in the detergent composition at a level of from 0.0001 % to 2% of enzyme protein by weight of the detergent composition, in parti- cular at a level of from 0.001 % to 0.1 % of enzyme protein by weight of the composition. Amylases: Any amylase (a and/or b) suitable for use in alkaline solutions can be used. Suitable amylases include those of bacterial or fungal origin. ChemicaUy or genetically modified mutants are included. Amylases include, for example, a-amylases obtained from a special strain of B. licheniformis. described in more detail in GB 1,296,839. Commer- cially avaUable amylases are Duramyl™, Termamyl™, Fungamyl™ and BAN™ (available from Novo Nordisk A/S) and Rapidase™ and Maxamyl P™ (available from (Gist-Brocades/Genencor) . The amylases are normally incoφorated in the detergent composition at a level of from 0.0001 % to 2% of enzyme protein by weight of the detergent composition, in parti- cular at a level of from 0.001 % to 0.1 % of enzyme protein by weight of the composition. Cellulases: Any cellulase suitable for use in alkaline solutions can be used. Suitable cel¬ lulases include those of bacterial or fungal origin. Chemically or genetically modified mutants are included. Suitable cellulases are disclosed in US 4,435,307, which discloses fungal cellulases produced from Humicola insolens. Especially suitable cellulases are the cellulases having color care benefits. Examples of such cellulases are cellulases described in published European patent application No. 0495257. Commercially available cellulases is Celluzyme® produced by a strain of Humicola insolens. (Novo Nordisk A/S), and KAC-500(B)® (Kao Coφoration). Said cellulases are normally incoφorated in the detergent composition at a level of from 0.0001 % to 2% of enzyme protein by weight of the detergent composition, in particular at a level of from 0.001 % to 0.1 % of enzyme protein by weight of the composition. Peroxidases/Oxidases : Peroxidase and/or oxidase enzymes, such as laccase, may be used, the former in combination with hydrogen peroxide sources, e.g., percarbonate, perbora¬ te, or persulfate, for solution bleaching, i.e. to prevent transfer of textile dyes from dyed fabrics to other fabrics when said fabrics are washed together in a wash liquor, preferably together with an enhancing agent as described in e.g. WO 94/12621 and WO 95/01426. Suitable enzymes of these types include those of plant, bacterial or fungal origin. Chemi¬ cally or genetically modified mutants are included. Said peroxidase and/or oxidase enzymes may be incoφorated in the detergent com- position at a level of from 0.0001 % to 2% of enzyme protein by weight of the detergent composition, in particular at a level of from 0.001 % to 0.1 % of enzyme protein by weight of the composition. Mixtures of the above mentioned enzymes are encompassed herein, in particular a mixture of a protease, an amylase, a lipase and/or a cellulase. Optional detergent ingredients Bleaching agents: Additional optional detergent ingredients that can be included in the detergent compositions of the present invention include peroxide bleaching agents such as sodium perborate-water (1/1), PB1, sodium perborate-water (1/4), PB4 and sodium car- bonate-hydrogen peroxide 2/3, percarbonate, with a particle size of 400-800 microns. If included, peroxide bleaching agents will typically be present at levels of from about 1 % to about 25%. Other peroxide-based bleaching agents that can be used are percarboxytic acids and salts thereof. Suitable examples of this class of agents include magnesium monoperoxy- phthalate hexahydrate, the magnesium salt of meta-chloro-perbenzoic acid, 4- nonylamino-4-oxoperoxybutyric acid and diperoxydodecanedioic acid. Such bleaching agents are disclosed in US 4,483,781, US 740,446, EP 0 133 354 and US 4,412,934. Highly preferred bleaching agents also include 6-nonylamino-6-oxoperoxycaproic acid as described in US 4,634,551. Bleaching agents for use herein may also be other bleaching agents known in the art for use in detergent compositions. A non-aqeous liquid detergent composition may comprise a peroxyacid material e.g peroxyacids consisting of N,N'-Di(4-Percarboxybenzoyl)ethylenediamine (PCBED), N,N'-Terephthaloyl-di(6-aminopercarboxycaproic acid) (TPCAP), N,N'-Di(4-percar- boxybenzoyl)piperazine (PCBPIP), N,N'-Di(4-Percarboxybenzoyl)-l,4- diaminocyclohexane (PCBHEX), N,N'-Di(4-Percarboxybenzoyl)-l ,4.butanediamine (PCBBD), N,N'-Di(4-Percarboxyaniline)-terephthalate (DPCAT), N.N.N'N'-l, 2,4,5- tetracarboxybenzoyl-di(6-aminopercarboxycaproic acid) (DiPAP), N,N'-Di(per- carboxyadipoyl)phenylenediamine (DPAPD) , N,N' -Succinoyl-di(4-percarboxy)aniUne (SDPCA), C 3 analogue of N,N'-Terephthaloyl-di-(8-amino peroxyoctanoic acid) (TPOCT) as described in WO 95/06104. Another such category of bleaching agents that can be used encompasses the halo¬ gen bleaching agents. Examples of hypohalite-releasing bleaching agents, for example, include trichloro-isocyanuric acid and the sodium and potassium dichloroisocyanurates and N-chloro and N-bromo alkanesulfonamides. Such materials are normally added at 0.5-10% by weight of the finished product, preferably 1-5% by weight. A further type of non-peroxidase based bleaching agent of particular interest inclu¬ des photoactivated bleaching agents such as the sulfonated zinc and/or aluminum phthalo- cyanines. These materials can be deposited upon the substrate during the washing pro¬ cess. Upon irradiation with light, in the presence of oxygen, such as by hanging clothes out to dry in the dayUght, the sulfonated phthalocyanine complex is activated and, conse- quently, the substrate is bleached. Prefened zinc phthalocyanine complexes and a pho¬ toactivated bleaching process are described in US 4,033,718. Typically, detergent com¬ positions will contain about 0.025% to about 1.25%, by weight, of sulfonated zinc phthalocyanine. The peroxide-based bleaching agents can be used in combination with bleach acti- vators such as tetraacetylethylenediamine (TAED), nonanoyloxybenzenesulfonate (NOBS, described in US 4,412,934), 3,5,5-trimethylhexanoyloxybenzenesulfonate (ISONOBS, described in EP 120 591) or pentaacetylglucose (PAG), which are perhydrolyzed to form a peracid as the active bleaching species, leading to improved bleaching effect. In addi¬ tion, very suitable are the bleach activators 6-octanamidocaproyloxybenzenesulfonate, 6- nonanamidocaproyl)oxybenzenesulfonate and 6-decanamidocaproyloxybenzenesulfonate or mixtures thereof. Also suitable activators are acylated citrate esters such as disclosed in European Patent Application No. 91870207.7. Further useful bleaching agents, including peroxyacids and bleaching systems com¬ prising bleach activators and peroxygen bleaching compounds for use in cleaning compo- sitions according to the invention, are described in application USSN 08/136,626. Hydrogen peroxide may also be generated in the detergent solution by adding an enzymatic system (i.e. an enzyme and a substrate therefore) which is capable of generati- SUBSTITUTE SHEET on of hydrogen peroxide at the beginning of or during the washing and/or rinsing pro¬ cess. Such enzymatic systems are disclosed in published EP Patent AppUcation No 0537381. The release of peracid from a peroxygen bleach source may be activated by use of a Upase of the invention. The necessary components for the enzymatic hydrolysis system is the peracid precursor substrate: a diacyl peroxide R 1 -CO-O-O-CO-R, where R and Rj may be saturated or unsaturated alkyl, an aryl group or an alkaryl. The lipase reacts with the substrate and releases peracids in the wash. Imine quatemary salts may used as bleach catalysts together with a peroxygen com- pound as described in WO 95/13352. Peroxide-based bleaching systems may also comprise a manganese catalyst. The manganese catalyst may, e.g., be one of the compounds described in "Efficient mangane¬ se catalysts for low-temperature bleaching", Nature 369, 1994, pp. 637-639. Suds suppressors: Another optional ingredient is a suds suppressor, exempUfied by siUco- nes, and silica-sUicone mixtures. Silicones can be generally represented by alkylated po- lysUoxane materials while silica is normally used in finely divided forms exemplified by silica aerogels and xerogels and hydrophobic siticas of various types. Theses materials can be incoφorated as particulates in which the suds suppressor is advantageously relea¬ sably incoφorated in a water-soluble or waterdispersible, substantially non surface-active detergent impermeable carrier. Altematively the suds suppressor can be dissolved or dis¬ persed in a liquid carrier and applied by spraying on to one or more of the other compo¬ nents. A preferred silicone suds controlling agent is disclosed in US 3,933,672. Other particularly useful suds suppressors are the self-emulsifying silicone suds suppressors, described in DE 2,646,126. An example of such a compound is DC-544, commercially avaUable form Dow Coming, which is a siloxane-glycol copolymer. Especially preferred suds controlUng agent are the suds suppressor system comprising a mixture of silicone oUs and 2-alkyl-alkanols. Suitable 2-alkyl-alkanols are 2-butyl-octanol which are com¬ mercially avaUable under the trade name Isofol 12 R. Such suds suppressor system are described in published European Patent AppUcati¬ on No. 0593841. Especially preferred sihcone suds controlling agents are described in pubUshed European Patent Application No. 0573699. Said compositions can comprise a sUicone/ silica mixture in combination with fumed nonporous silica such as Aerosil R . The suds suppressors described above are normally employed at levels of from 0.001 % to 2% by weight of the composition, preferably from 0.01 % to 1 % by weight. Other components: Other components used in detergent compositions may be employed such as soU-suspending agents, soil-releasing agents, optical brighteners, abrasives, bac¬ tericides, tarnish inhibitors, coloring agents, and/or encapsulated or nonencapsulated per¬ fumes. Especially suitable encapsulating materials are water soluble capsules which consist of a matrix of polysaccharide and polyhydroxy compounds such as described in GB 1,464,616. Other suitable water soluble encapsulating materials comprise dextrins derived from ungelatinized starch acid esters of substituted dicarboxylic acids such as described in US 3,455,838. These acid-ester dextrins are, preferably, prepared from such starches as wa¬ xy maize, waxy sorghum, sago, tapioca and potato. Suitable examples of said encapsula¬ tion materials include N-Lok manufactured by National Starch. The N-Lok encapsulating material consists of a modified maize starch and glucose. The starch is modified by ad¬ ding monofunctional substituted groups such as octenyl succinic acid anhydride. Antiredeposition and soil suspension agents suitable herein include cellulose deriva¬ tives such as methylcellulose, carboxymethylcellulose and hydroxyethylcellulose, and homo- or co-polymeric polycarboxylic acids or their salts. Polymers of this type include the polyacrylates and maleic anhydride-acrylic acid copolymers, e.g Sokalan CP5, pre¬ viously mentioned as builders, as well as copolymers of maleic anhydride with ethylene, methylvinyl ether or methacrylic acid, the maleic anhydride constituting at least 20 mole percent of the copolymer. These materials are normally used at levels of from 0.5% to 10% by weight, more preferably form 0.75% to 8%, most preferably from 1% to 6% by weight of the composition. Preferred optical brighteners are anionic in character, examples of which are disodium 4,4'-bis-(2-diethanolamino-4-anilino -s- triazin-6-ylamino)stilbene-2:2' disulphonate, dis¬ odium 4, - 4 l -bis-(2-moφholino-4-anilino-s-triazin-6-ylamino-stUbene-2:2' - disulphona- te, disodium 4,4' - bis-(2,4-dianUino-s-triazin-6-ylamino)stilbene-2:2' - disulphonate, monosodium 4', 4" - bis-(2,4-diamlino-s-tri-azm-6 ylamino)stilbene-2-sulphonate, diso¬ dium 4,4 ' -bis-(2-anilino-4-(N-methyl-N-2-hydroxyethylamino)-s-triazin-6- ylamino)stilbene-2,2' - disulphonate, di-sodium 4,4' -bis-(4-phenyl-2,l,3-triazol-2-yl)- 5 stilbene-2,2' disulphonate, di-so-dium 4,4 , bis(2-aniUno-4-(l-methyl-2-hy- droxyethylamino)-s-triazin-6-ylami-no)stilbene-2,2 l disulphonate, sodium 2(stilbyl-4' '- (naphtho-l',2':4,5)-l,2,3, - triazole-2" -sulphonate and 4,4'-bis(2-sulphostyryl)biphenyl. Other useful polymeric materials are the polyethylene glycols, particularly those of molecular weight 1000-10000, more particularly 2000 to 8000 and most preferably about 10 4000. These are used at levels of from 0.20% to 5% more preferably from 0.25% to 2.5% by weight. These polymers and the previously mentioned homo- or co-polymeric polycarboxylate salts are valuable for improving whiteness maintenance, fabric ash depo¬ sition, and cleaning performance on clay, proteinaceous and oxidizable soUs in the pre¬ sence of transition metal impurities. 15 A graft polymer as described in WO 95/22593 may also be used. Soil release agents useful in compositions of the present invention are conventio¬ nally copolymers or teφolymers of terephthalic acid with ethylene glycol and/or propyle¬ ne glycol units in various arrangements. Examples of such polymers are disclosed in US 4,116,885, US 4,711,730 and EP 0 272 033. A particular preferred polymer in accor- 0 dance with EP 0 272 033 has the formula: (CH 3 (PEG) 43 )o. 7 s(POH) 0 . 25 [T-PO) 2 . 8 (T-PEG) 0 . 4 ]T(POH) 0 . 25 ((PEG) 4 3CH3)o. 7 5 where PEG is -(OC 2 H 4 )0-, PO is (OC 3 H 6 O) and T is (pcOC 6 H 4 CO). Also very useful are modified polyesters as random copolymers of dimethyl te- 5 rephthalate, dimethyl sulfoisophthalate, ethylene glycol and 1-2 propane diol, the end groups consisting primarily of sulphobenzoate and secondarily of mono esters of ethylene glycol and/or propane-diol. The target is to obtain a polymer capped at both end by sulp¬ hobenzoate groups, "primarily", in the present context most of said copolymers herein wiU be endcapped by sulphobenzoate groups. However, some copolymers will be less 0 than fuUy capped, and therefore their end groups may consist of monoester of ethylene glycol and/or propane 1-2 diol, thereof consist "secondarily" of such species. The selected polyesters herein contain about 46% by weight of dimethyl terephtha- Uc acid, about 16% by weight of propane -1.2 diol, about 10% by weight ethylene gluy- col about 13% by weight of dimethyl sulfobenzoic acid and about 15% by weight of sul- foisophthalic acid, and have a molecular weight of about 3.000. The polyesters and their method of preparation are described in detail in EP 311 342. Softening agents: Fabric softening agents can also be incoφorated into laundry detergent compositions in accordance with the present invention. These agents may be inorganic or organic in type. Inorganic softening agents are exemplified by the smectite clays disclo¬ sed in GB-A-1 400898 and in US 5,019,292. Organic fabric softening agents include the water insoluble tertiary amines as disclosed in GB-A1 514 276 and EP 0 011 340 and their combination with mono C ι2 -C 14 quatemary ammonium salts are disclosed in EP 026 528 and di-long-chain amides as disclosed in EP 0 242 919. Other useful organic ingredi¬ ents of fabric softening systems include high molecular weight polyethylene oxide materi¬ als as disclosed in EP 0 299 575 and 0 313 146. Levels of smectite clay are normally in the range form 5 % to 15 % , more preferably form 8% to 12% by weight, with the material being added as a dry mixed component to the remainder of the formulation. Organic fabric softening agents such as the water- insoluble tertiary amines or dilong chain amide materials are incoφorated at levels of from 0.5% to 5% by weight, normally from 1 % to 3% by weight whilst the high mo- lecular weight polyethylene oxide materials and the water soluble cationic materials are added at levels of from 0.1 % to 2%, normally from 0.15% to 1.5% by weight. These materials are normally added to the spray dried portion of the composition, although in some instances it may be more convenient to add them as a dry mixed particulate, or spray them as molten liquid on to other solid components of the composition. Polymeric dye transfer inhibiting agents: The detergent compositions according to the present invention may also comprise from 0.001% to 10%, preferably from 0.01% to 2%, more preferably form 0.05% to 1% by weight of polymeric dye transfer inhibiting agents. Said polymeric dye transfer inhibiting agents are normally incoφorated into de¬ tergent compositions in order to inhibit the transfer of dyes from colored fabrics onto fa- brics washed therewith. These polymers have the abUity to complex or adsorb the fugitive dyes washed out of dyed fabrics before the dyes have the opportunity to become attached to other articles in the wash. Especially suitable polymeric dye transfer inhibiting agents are polyamine N-oxide polymers, copolymers of N-vinylpyrroUdone and N-vinylimidazole, polyvmylpyrroUdone polymers, polyvinyloxazolidones and polyvinylimidazoles or mixtures thereof. Addition of such polymers also enhances the performance of the enzymes according the invention. The detergent composition according to the invention can be in liquid, paste, gels, bars or granular forms. Non-dusting granulates may be produced, e.g., as disclosed in US 4,106,991 and 4,661,452 (both to Novo Industri A/S) and may optionaUy be coated by methods known in the art. Examples of waxy coating materials are poly(ethylene oxi¬ de) products (polyethyleneglycol, PEG) with mean molecular weights of 1000 to 20000; ethoxylated nonylphenols having from 16 to 50 ethylene oxide units; ethoxylated fatty alcohols in which the alcohol contains from 12 to 20 carbon atoms and in which there are 15 to 80 ethylene oxide units; fatty alcohols; fatty acids; and mono- and di- and triglyce¬ rides of fatty acids. Examples of film-forming coating materials suitable for application by fluid bed techniques are given in GB 1483591. Granular compositions according to the present invention can also be in "compact form", i.e. they may have a relatively higher density than conventional granular deter¬ gents, i.e. form 550 to 950 g/1; in such case, the granular detergent compositions accor¬ ding to the present invention will contain a lower amount of "Inorganic filler salt", com¬ pared to conventional granular detergents; typical filler salts are alkaline earth metal salts of sulphates and chlorides, typically sodium sulphate; "Compact" detergent typically comprise not more than 10% filler salt. The liquid compositions according to the present invention can also be in "concentrated form", in such case, the liquid detergent composi¬ tions according to the present invention will contain a lower amount of water, compared to conventional liquid detergents. Typically, the water content of the concentrated liquid detergent is less than 30%, more preferably less than 20%, most preferably less than 10% by weight of the detergent compositions. The compositions of the invention may for example, be formulated as hand and ma¬ chine laundry detergent compositions including laundry additive compositions and com¬ positions suitable for use in the pretreatment of stained fabrics, rinse added fabric softe- ner compositions, and compositions for use in general household hard surface cleaning operations and dishwashing operations. Particular forms of laundry detergent compositions within the invention include: 1) A detergent composition formulated as a granulate having a bulk density of at least 5 600 g/1 comprising Linear alkylbenzenesulfonate (calculated as acid) 7 - 12% Alcohol ethoxysulfate (e.g. Cι 2 -ι 8 alcohol, 1-2 EO) or alkyl sulfate (e.g. C 16 ., 8 ) 1 - 4% Alcohol ethoxylate (e.g. C l4 _ l5 alcohol, 7 EO) 5 - 9% Sodium carbonate (as Na 2 CO 3 ) 14 - 20% Soluble silicate (as Na 2 O,2SiO 2 ) 2 - 6% Zeolite (as NaAlSiO 4 ) 15 - 22% Sodium sulfate (as Na 2 SO 4 ) 0 - 6% Sodium citrate/citric acid (as C 6 H 5 Na 3 O 7 /C 6 H 8 O 7 ) 0 - 15% Sodium perborate (as NaBO 3 .H 2 O) 11 - 18% TAED 2 - 6% Carboxymethylcellulose 0 - 2% Polymers (e.g. maleic/acrylic acid copolymer, PVP, PEG) 0 - 3% Enzymes (calculated as pure enzyme 0.0001 - 0.1% protein) Minor ingredients (e.g. suds suppressors, perfume, optical brightener, photobleach) 0 - 5% 2) A detergent composition formulated as a granulate having a bulk density of at least 600 g/1 comprising Linear alkylbenzenesulfonate (calculated as acid) 6 - 11 % Alcohol ethoxysulfate (e.g. C 8 alcohol, 1-2 EO or alkyl sulfate (e.g. C 16 . 18 ) 1 - 3% Alcohol ethoxylate (e.g. C^.^ alcohol, 7 EO) 5 - 9% Sodium carbonate (as Na 2 CO 3 ) 15 - 21% Soluble siUcate (as Na 2 O,2Siθ 2 ) 1 - 4% ZeoUte (as NaAlSiO 4 ) 24 - 34% Sodium sulfate (as Na 2 SO 4 ) 4 - 10% Sodium citrate/citric acid (as C 6 H 5 Na 3 O 7 /C 6 H 8 O 7 ) 0 - 15% CarboxymethylceUulose 0 - 2% Polymers (e.g. maleic/acrylic acid copolymer, PVP, PEG) 1 - 6% Enzymes (calculated as pure enzyme 0.0001 - 0.1% protein) Minor ingredients (e.g. suds suppressors, perfume) 0 - 5% 3) A detergent composition formulated as a granulate having a bulk density of at least 600 g/1 comprising Linear alkylbenzenesulfonate (calculated as acid) 5 - 9% Alcohol ethoxylate (e.g. C 5 alcohol, 7 EO) 7 - 14% Soap as fatty acid (e.g. C 16 . 22 fatty acid) 1 - 3% Sodium carbonate (as Na 2 CO 3 ) 10 - 17% Soluble silicate (as Na 2 O,2SiO 2 ) 3 - 9% Zeolite (as NaAlSiO 4 ) 23 - 33% Sodium sulfate (as Na 2 SO4) 0 - 4% Sodium perborate (as NaBO 3 .H 2 O) 8 - 16% TAED 2 - 8% Phosphonate (e.g. EDTMPA) 0 - 1% Carboxymethylcellulose 0 - 2% Polymers (e.g. maleic/acrylic acid copolymer, PVP, PEG) 0 - 3% Enzymes (calculated as pure enzyme 0.0001 - 0.1 % protein) Minor ingredients (e.g. suds suppressors, perfume, optical brightener) 0 - 5% 4) A detergent composition formulated as a granulate having a bulk density of at least 600 g/1 comprising Linear alkylbenzenesulfonate (calculated as acid) 8 - 12% Alcohol ethoxylate (e.g. C 12 . 15 alcohol, 7 EO) 10 - 25% Sodium carbonate (as Na 2 CO 3 ) 14 - 22% Soluble silicate (as Na 2 θ,2SiO 2 ) 1 - 5% ZeoUte (as NaAlSiO 4 ) 25 - 35% Sodium sulfate (as Na^O.,) 0 - 10% Carboxymethylcellulose 0 - 2% Polymers (e.g. maleic/acrylic acid copolymer, PVP, PEG) 1 - 3% Enzymes (calculated as pure enzyme 0.0001 - 0.1 % protein) Minor ingredients (e.g. suds suppressors, perfume) 0 - 5% 5) An aqueous liquid detergent composition comprising Linear alkylbenzenesulfonate (calculated as acid) 15 - 21% Alcohol ethoxylate (e.g. C 12 . 15 alcohol, 7 EO or C, 2 . 15 alcohol, 5 EO) 12 - 18% Soap as fatty acid (e.g. oleic acid) 3 - 13% Alkenylsuccinic acid (C, 2 ., 4 ) 0 - 13% Aminoethanol 8 - 18% Citric acid 2 - 8% Phosphonate 0 - 3% Polymers (e.g. PVP, PEG) 0 - 3% Borate (as B 4 O 7 ) 0 - 2% Ethanol 0 - 3% Propylene glycol 8 - 14% Enzymes (calculated as pure enzyme 0.0001 - 0.1% protein) Minor ingredients (e.g. dispersants, suds suppressors, perfume, optical brightener) 0 - 5% 6) An aqueous stmctured liquid detergent composition comprising Linear alkylbenzenesulfonate (calculated as acid) 15 - 21% Alcohol ethoxylate (e.g. C 12 ., 5 alcohol, 7 EO, or C 12 . 15 alcohol, 5 EO) 3 - 9% Soap as fatty acid (e.g. oleic acid) 3 - 10% ZeoUte (as NaAlSiO 4 ) 14 - 22% Potassium citrate 9 - 18% Borate (as B 4 O 7 ) 0 - 2% Carboxymethylcellulose 0 - 2% Polymers (e.g. PEG, PVP) 0 - 3% Anchoring polymers such as, e.g., lauryl methacrylate/acrylic acid copolymer; molar ratio 0 - 3% 25:1; MW 3800 Glycerol 0 - 5% Enzymes (calculated as pure enzyme protein) 0.0001 - 0.1% Minor ingredients (e.g. dispersants, suds suppressors, perfume, optical brighteners) 0 - 5% 7) A detergent composition formulated as a granulate having a bulk density of at least 600 g/1 comprising Fatty alcohol sulfate 5 - 10% Ethoxylated fatty acid monoethanolamide 3 - 9% Soap as fatty acid 0 - 3% Sodium carbonate (as Na 2 CO 3 ) 5 - 10% Soluble sUicate (as Na 2 O,2SiO 2 ) 1 - 4% Zeolite (as NaAlSiO 4 ) 20 - 40% Sodium sulfate (as Na 2 SO 4 ) 2 - 8% Sodium perborate (as NaBO 3 .H 2 O) 12 - 18% TAED 2 - 7% Polymers (e.g. maleic/acrylic acid copolymer, PEG) 1 - 5% Enzymes (calculated as pure enzyme 0.0001 - 0.1% protein) Minor ingredients (e.g. optical brightener, suds suppressors, perfume) 0 - 5% TE SHEET 8) A detergent composition formulated as a granulate comprising Linear alkylbenzenesulfonate (calculated as acid) 8 - 14% Ethoxylated fatty acid monoethanolamide 5 - 11% Soap as fatty acid 0 - 3% Sodium carbonate (as Na 2 CO 3 ) 4 - 10% Soluble silicate (as Na 2 O,2SiO 2 ) 1 - 4% Zeolite (as NaAlSiO 4 ) 30 - 50% Sodium sulfate (as Na 2 SO 4 ) 3 - 11% Sodium citrate (as C 6 H 5 Na 3 O 7 ) 5 - 12% Polymers (e.g. PVP, maleic/acrylic acid 1 - 5% copolymer, PEG) Enzymes (calculated as pure enzyme 0.0001 - 0.1% protein) Minor ingredients (e.g. suds suppressors, perfume) 0 - 5% SUBSTITUTE SHEET 9) A detergent composition formulated as granulate Alkyl sulfate (e.g C 12 .,„) 6 - 12 % Soap, Na-salt 0-3 % Nonionic, (eg. alcohol ethoxylate C iQ . ls , 2- 2-8% 7EO) Alkylglucamid (e.g C 16 . 18 ) 2-6% ZeoUte (NaAlSiO 4 ) 14 - 24 % Sodium carbonate (Na 2 CO 3 ) 7 - 13 % Sodium disiUicate; (Na 2 O:2SiO 2 ) 10 - 14 % (e.g SKS6) Sodium sulfate (Na 2 SO 4 ) 5-9% Sodium percarbonate 10-16% TAED 1-5% CMC 0-3 % Polycarboxylate 1-7% Polymers (e.g PVP, PEG, maleic/acrylic 0-2% acid copolymer) Enzymes including Upolytic enzymes 0.0001-0.1 % (calculated as pure enzyme protein) Minor ingrediens (e.g suds supressors, 0-5 % perfume, optical brightener, photobleach) Bulk density (g/1) at least 700 10) A detergent composition formulated as granulate Alkyl sulfate (e.g C 12 .ι 8 ) 7-11 % Soap, Na-salt 0-3 % Nonionic, (eg. alcohol ethoxylate C 10 ., 8 , 2- 7-11 % 7EO) Alkylglucamid (e.g Ci 6 ., 8 ) 2-6% ZeoUte (NaAlSiO 4 ) 19 - 29 % Sodium carbonate (Na 2 CO 3 ) 12 - 18 % Sodium disillicate; (Na 2 O:2Siθ 2 ) 7-11 % (e.g SKS6) Sodium sulfate (Na^O,) 5-9 % CMC 0-3 % Polycarboxylate 4-8 % Polymers (e.g PVP, PEG, maleic/acrylic 1-3 % acid copolymer) Enzymes including Upolytic enzymes 0.0001-0.1 % (calculated as pure enzyme protein) Minor ingrediens (e.g suds supressors, 0-5 % perfume, optical brightener, photobleach) Bulk density (g/1) at least 700 11) A detergent composition formulated as granulate Alkyl sulfate (e.g Cι 2 .ι 8 ) 4 - 10 % Nonionic, (eg. alcohol ethoxylate Cιo.ι 8 , 2- 2-7% 7EO) Phosphate (as STPP) 17 - 27 % Sodium carbonate (Na 2 CO 3 ) 4-8% Sodium disilUcate; (Na 2 O:2SiO 2 ) 4-8% (e.g SKS6) Sodium sulfate (Na 2 SO 4 ) 18 - 26 % Sodium perborate tetrahydrate 13 - 19 % TAED 1-4 % CMC 0-2% Enzymes including Upolytic enzymes 0.0001-0.1 % (calculated as pure enzyme protein) Minor ingrediens (e.g suds supressors, 0-5 % perfume, optical brightener, photobleach) Bulk density (g/1) at least 600 12) A detergent composition formulated as granulate Alkyl sulfuric acid (e.g C 8 ) 2-9 % Soap, Na-salt 1-4 % Nonionic, (eg. alcohol ethoxylate C 10 ., g , 2- 9 - 15 % 7EO) ZeoUte (NaAlSiO 4 ) 35 - 45 % Sodium carbonate (Na 2 CO 3 ) 3 - 10 % Sodium disiUicate; (Na 2 O:2Siθ 2 ) 0-4 % (e.g SKS6) Sodium sulfate (Na 2 SO 4 ) 2-6% Sodium percarbonate 14 - 20 % TAED 2-7% CMC 0-3 % Polycarboxylate 0-2 % Enzymes including lipolytic enzymes 0.0001-0.1 (calculated as pure enzyme protein) Minor ingrediens (e.g suds supressors, 0-5 % perfume, optical brightener, photobleach) Bulk density (g/1) at least 700 13) A detergent composition formulated as granulate Alkyl benzene sulfonic acid 8 - 14 % Alkyl sulfuric acid (e.g C I2 . 18 ) 2-6% Nonionic, (eg. alcohol ethoxylate C 1(M8 , 2- 9 - 13 % 7EO) ZeoUte (NaAlSiO,) (e.g ZeoUte 4A) 20 - 30 % Sodium carbonate (Na 2 CO 3 ) 0-6% Sodium disiUicate; (Na 2 O:2SiO 2 ) 0-3 % Sodium sulfate (Na 2 SO 4 ) 0-6 % Sodium perborate tetrahydrate 22 - 28 % TAED 5-9% CMC 0-2 % Polymers (e.g maleic/acrylic acid 0-4% copolymer) Enzymes including Upolytic enzymes 0.0001-0.1 (calculated as pure enzyme protein) Minor ingrediens (e.g suds supressors, 0-5% perfume, optical brightener, photobleach) Bulk density (g/1) at least 600 14) A detergent composition formulated as granulate Alkyl benzene sulfonic acid 6 - 12 % Alkyl ether sulfuric acid (e.g. Cι 2 . 18 2-6 % alcohol, 4-10 EO) or alkyl sulfate (e.g C 12 . u) Soap, Na-salt 0-2 % Nonionic, (eg. alcohol ethoxylate C 12 .ι 8 , 3- 9 - 13 % 10 EO) Zeolite (NaAlSiO 4 ) (e.g Zeolite 4A) 39 - 49 % Sodium carbonate (Na 2 CO 3 ) 2-8 % Sodium disillicate; (Na 2 O:2SiO 2 ) 0-3 % Sodium sulfate (Na 2 SO 4 ) 2-8 % CMC 0- 3 % Polymers (e.g maleic/acrylic acid 0-4 % copolymer) Enzymes including lipolytic enzymes 0.0001-0.1 (calculated as pure enzyme protein) Minor ingrediens (e.g suds supressors, 0-5 % perfume, optical brightener, photobleach) Bulk density (g/1) at least 600 SUBSTITUTE SHEET 15) A detergent composition formulated as granulate Alkyl sulfate (e.g C 12 -ι 8 ) 2-8 % Soap, Na-salt 0-3 % Nonionic, (eg. alcohol ethoxylate C 6 , 3- 10 - 16 % 10 EO) ZeoUte (NaAlSiO,) (e.g ZeoUte 4A) 47 - 57 % Sodium carbonate (Na 2 CO 3 ) 15 - 23 % Sodium citrate /citric acid 0 - 16 % (C 6 H 5 Na 3 O 7 /C 6 H 8 O 7 ) Sodium sulfate (Na 2 SO 4 ) 1-5 % CMC 0-3% Polycarboxylate 0-2 % Polymers (e.g PVP) 0-2 % Enzymes including lipolytic enzymes 0.0001-0.1 (calculated as pure enzyme protein) Minor ingrediens (e.g suds supressors, 0-5 % perfume, optical brightener, photobleach) Bulk density (g/1) at least 800 SUBSTITUTE SHEET 16) A detergent composition formulated as granulate SUBSTITUTE SHEET 17) A detergent composition formulated as granulate Soap, Na-salt 1-3 % Nonionic, (eg. alcohol ethoxylate C 12 . 18 , 3- 2-6% 10 EO) Betaine (e.g Alkylamidopropylbetaine) 0-3 % Phosphate (as STPP) 27 - 37 % Sodium carbonate (Na 2 CO 3 ) 17 - 23 % Sodium disUlicate; (Na 2 O:2SiO 2 ) 3-7 % Sodium sulfate (Na 2 SO 4 ) 4-11 % Sodium perborate tetrahydrate 15-21 % TAED 1-4 % CMC 1-3 % Enzymes including lipolytic enzymes 0.0001-0.1 (calculated as pure enzyme protein) Minor ingrediens (e.g suds supressors, 0-5 % perfume, optical brightener, photobleach) Bulk density (g/1) at least 600 SUBSTITUTE SHEET 18) A detergent composition formulated as granulate Alkyl benzene sulfonic acid 5-11 % Soap, Na-salt 0-3 % Nonionic, (eg. alcohol ethoxylate C 12 .ι 8 , 3- 3-7% 10 EO) Zeolite (NaAlSiO 4 ) (e.g ZeoUte 4A) 20 - 30 % Sodium carbonate (Na 2 CO 3 ) 15 - 23 % Sodium citrate /citric acid 0-3 % (C 6 H s Na 3 O 7 /C 6 H 8 O 7 ) Sodium sulfate (Na 2 SO 4 ) 4 - 10 % Sodium percarbonate 7 -13 % TAED 2-6 % CMC 1-3 % Enzymes including lipolytic enzymes 0.0001-0.1 (calculated as pure enzyme protein) Minor ingrediens (e.g suds supressors, 0-5 % perfume, optical brightener, photobleach) Bulk density (g/1) at least 600 19) A detergent composition formulated as granulate Alkyl benzene sulfonic acid 0-4% Soap, Na-salt 0-3 % Nonionic, (eg. alcohol ethoxylate Cι 2 . 18 , 3- 2-6 % 10 EO) Zeolite (NaAlSiO 4 ) (e.g Zeolite 4A) 11 - 17 % Phosphate (as STPP) 25 - 35 % Sodium carbonate (Na 2 CO 3 ) 3-7% Sodium silUcate; (Na 2 O:SiO 2 ) 0 - 19 % Sodium sulfate (Na 2 SO 4 ) 20 - 28 % Sodium perborate tetrahydrate 9 - 13 % TAED 1-5 % CMC 0-2% Enzymes including lipolytic enzymes 0.0001-0.1 (calculated as pure enzyme protein) Minor ingrediens (e.g suds supressors, 0-5 % perfume, optical brightener, photobleach) Bulk density (g/1) at least 600 20) A detergent composition formulated as granulate Alkyl benzene sulfonic acid 17 - 23 % Soap, Na-salt 0-3 % Nonionic, (eg. alcohol ethoxylate Cι 2 . 18 , 3- 11 - 15 % 10 EO) ZeoUte (NaAlSiO 4 ) (e.g Zeolite 4A) 60 - 70 % Sodium carbonate (Na 2 CO 3 ) 0-3 % Sodium sulfate (Na 2 SO 4 ) 5 - 11 % CMC 0-3 % Polymers (e.g PVP, PEG, maleic/acrylic 2-6% acid copolymer)) Enzymes including lipolytic enzymes 0.0001-0.1 (calculated as pure enzyme protem) Minor ingrediens (e.g suds supressors, 0-5 % perfume, optical brightener, photobleach) Bulk density (g/1) at least 350 21) A detergent composition formulated as granulate Alkyl benzene sulfonic acid 16 - 22 % Soap, Na-salt 0-2% Nonionic, (eg. alcohol ethoxylate C 8 , 3- 3-9% 10 EO) ZeoUte (NaAlSiO 4 ) 25 - 33 % Sodium carbonate (Na 2 CO 3 ) 3-7 % Sodium sillicate; ' (Na 2 O:SiO 2 ) 0 - 4 % Sodium sulfate (Na 2 SO 4 ) 5-11 % Phosphonate 0-3 % Sodium perborate monohydrate 15 - 19 % TAED 3-7% CMC 0-3 % Polymers (e.g PVP, PEG, maleic/acrylic 0-3 % acid copolymer)) Enzymes including lipolytic enzymes 0.0001-0.1 (calculated as pure enzyme protein) Minor ingrediens (e.g suds supressors, 0-5 % perfume, optical brightener, photobleach) Bulk density (g/1) at least 700 22) A detergent composition formulated as granulate Alkyl benzene sulfonic acid 4-8 % Alkyl sulfate (e.g C 8 ) 0-3% Nonionic, (eg. alcohol ethoxylate C 12 .ι 8 , 3- 5-9 % 10 EO) Zeolite (NaAlSiO 4 ) 20 - 28 % Sodium carbonate (Na 2 CO 3 ) 9 - 15 % Sodium disillicate; (Na 2 O:2SiO 2 ) 0-4 % Sodium perborate tetrahydrate 21 - 31 % TAED 1-5 % CMC 0-3 % Polymers (e.g PVP, PEG, maleic/acrylic 0-3 % acid copolymer)) Enzymes including lipolytic enzymes 0.0001-0.1 (calculated as pure enzyme protein) Minor ingrediens (e.g suds supressors, 0-5 % perfume, optical brightener, photobleach) Bulk density (g/1) at least , 600 1 BSTITUTE SHEET 23) A detergent composition formulated as a granulate comprising Linear alkylbenzenesulfonate (calculated as acid) 6 - 12% Nonionic surfactant 1 - 4% Soap as fatty acid 2 - 6% Sodium carbonate (as Na 2 CO 3 ) 14 - 22% Zeolite (as NaAlSiO 4 ) 18 - 32% Sodium sulfate (as Na 2 SO 4 ) 5 - 20% Sodium citrate (as C 6 H 5 Na 3 O 7 ) 3 - 8% Sodium perborate (as NaBO 3 .H 2 O) 4 - 9% Bleach activator (e.g. NOBS or TAED) 1 - 5% Carboxymethylcellulose 0 - 2% Polymers (e.g. polycarboxylate or PEG) 1 - 5% Enzymes (calculated as pure enzyme 0.0001 - 0.1% protein) Minor ingredients (e.g. optical brightener, perfume) 0 - 5% | T TE SHEET 24) An aqueous liquid detergent composition comprising Linear alkylbenzenesulfonate (calculated as acid) 15 - 23% Alcohol ethoxysulfate (e.g. C 12 .ι 5 alcohol, 2-3 EO) 8 - 15% Alcohol ethoxylate (e.g. C 12 . 15 alcohol, 7 EO, or Ci 2 . 15 alcohol, 5 EO) 3 - 9% Soap as fatty acid (e.g. lauric acid) 0 - 3% Aminoethanol 1 - 5% Sodium citrate 5 - 10% Hydrotrope (e.g. sodium toluensulfonate) 2 - 6% Borate (as B 4 O 7 ) 0 - 2% Carboxymethylcellulose 0 - 1 % Ethanol 1 - 3% Propylene glycol 2 - 5% Enzymes (calculated as pure enzyme protein) 0.0001 - 0.1% Minor ingredients (e.g. polymers, dispersants, perfume, optical brighteners) 0 - 5% 25) An aqueous liquid detergent composition comprising Linear alkylbenzenesulfonate (calculated as acid) 20 - 32% Alcohol ethoxylate (e.g. Cι 2 ., 5 alcohol, 7 EO, or C 12 . 15 alcohol, 5 EO) 6 - 12% Aminoethanol 2 - 6% Citric acid 8 - 14% Borate (as B 4 O 7 ) 1 - 3% Polymer (e.g. maleic/acrylic acid copolymer, anchoring polymer such as, e.g., lauryl methacrylate/acrylic acid copolymer) 0 - 3% Glycerol 3 - 8% Enzymes (calculated as pure enzyme 0.0001 - 0.1% protein) Minor ingredients (e.g. hydrotropes, dispersants, perfume, optical brighteners) 0 - 5% T 26) A detergent composition formulated as concentrated Uquid. Alkyl benzene sulfonic acid 6-12% Alkyl sulfuric acid 0-4% Monoethanolamin 2-6% Nonionic, (eg. alcohol ethoxylate C, 2 -ι 8 , 3- 9 - 15 % 10 EO) Sodium citrate /citric acid 2-8% (C^NaA/ H ) Glycerol 2-6% Borate (as Na 2 B 4 O 7 ) 0-4 % Polymers (e.g PVP, PEG, maleic/acrylic 0-3 % acid copolymer)) Enzymes including lipolytic enzymes 0.0001-0.1 (calculated as pure enzyme protein) Minor ingrediens (e.g suds supressors, 0-5 % perfume, optical brightener, photobleach) Total water 40% 27) A detergent composition formulated as concentrated liquid. Alkyl benzene sulfonic acid 11 - 17 % Alkyl ether sulfuric acid (e.g. Cι 2 . 18 0 - 4 % alcohol, 4-10 EO) or alkyl sulfate (e.g C 12 . ll) Triethanolamin 0 - 3 % Nonionic, (eg. alcohol ethoxylate C, 2 -ι 8 , 3- 6 - 10 % 10 EO) Sodium citrate /citric acid 2 - 6 % (C 6 H 5 Na 3 O 7 /C 6 H 8 O 7 ) Hydrotropes (Sodium toluene sulfonate) 1 - 5 % Glycerol 6 - 12 % MPG 0 - 5 % Ethanol 0 - 3 % Enzymes including Upolytic enzymes 0.0001 - 0.1 (calculated as pure enzyme protein) Minor ingrediens (e.g suds supressors, 0 - 5 % perfume, optical brightener, photobleach) Total water 55 % 28) A detergent composition formulated as a granulate having a bulk density of at least 600 g/1 comprising Anionic surfactant (linear alkylbenzenesulfonate, alkyl sulfate, alpha-olefinsulfonate, alpha-sulfo fatty acid methyl esters, alkanesulfonates, soap) 25 - 40% Nonionic surfactant (e.g. alcohol 1 - 10% ethoxylate) Sodium carbonate (as Na 2 CO 3 ) 8 - 25% Soluble silicates (as Na 2 O, 2SiO 2 ) 5 - 15% Sodium sulfate (as Na 2 SO 4 ) 0 - 5% ZeoUte (as NaAlSiO 4 ) 15 - 28% Sodium perborate (as NaBO 3 .4H 2 O) 0 - 20% Bleach activator (TAED or NOBS) 0 - 5% Enzymes (calculated as pure enzyme 0.0001 - 0.1 % protein) SUBSTITUTE SHEET Minor ingredients (e.g. perfume, optical 0 - 3% brighteners) 29) A detergent composition formulated as granulate Alkyl benzene sulfonic acid 25 - 35 % Nonionic, (eg. alcohol ethoxylate C 12 . 18 , 3- 0-3% 10 EO) ZeoUte (NaAlSiO,) 3-9% Phosphate (as STPP) 25 - 35 % Sodium carbonate (Na 2 CO 3 ) 0-3 % Sodium disillicate; (Na 2 O:2SiO 2 ) 2-8 % Sodium sulfate (Na 2 SO 4 ) 17 - 23 % Sodium perborate monohydrate 1-5 % TAED 0-3 % CMC 0-3 % Enzymes including Upolytic enzymes 0.0001-0.1 (calculated as pure enzyme protein) Minor ingrediens (e.g suds supressors, 0-5 % perfume, optical brightener, photobleach) Bulk density (g/1) at least 600 30) A detergent composition formulated as granulate Alkyl benzene sulfonic acid 25 - 35 % Soap, fatty acid Na-salt 0 - 3 % Nonionic, (eg. alcohol ethoxylate Cι 3 . J5 , 7 4 - 9 % EO) Zeolite (NaAlSiO 4 ) 7 - 11 % Phosphate (as STPP) 26 - 36 % Sodium carbonate (Na 2 CO 3 ) 6 - 12 % Sodium disillicate; (Na 2 O:2SiO 2 ) 4 - 10 % Sodium sulfate (Na 2 SO 4 ) 4 - 8 % CMC 0 - 3 % Enzymes including lipolytic enzymes 0.0001 - 0.1 (calculated as pure enzyme protein) Minor ingrediens (e.g suds supressors, 0 - 5 % perfume, optical brightener, photobleach) Bulk density (g/1) at least 700 The following specific compositions are meant to exemplify compositions for the present invention, but are not necessarily meant to limit or otherwise define the scope of the invention. In the detergent compositions, the abbreviated component identifications have the following meanings: LAS: Sodium linear C 12 alkyl benzene sulphonate TAS: Sodium tallow alkyl sulphate XYAS: Sodium C 1X - C 1Y alkyl sulfate SS: Secondary soap surfactant of formula 2-butyl octanoic acid 25EY: A C 12 - C 1S predominantly Unear primary alcohol condensed with an average of Y moles of ethylene oxide 45EY: A C M - C l5 predominantly Unear primary alcohol condensed with an average of Y moles of ethylene oxide TITUTE SHEET XYEZS: C 1X - C 1Y sodium alkyl sulfate condensed with an average of Z moles of ethylene oxide per mole Nonionic: C 13 - C ϊS mixed ethoxylated/propoxylated fatty alcohol with an average degree of ethoxylation of 3.8 and an average degree of propoxylation of 4.5 sold under the tradename Plurafax LF404 by BASF Gmbh CFAA: Cj 2 - C M alkyl N-methyl glucamide TFAA: Cι 6 - C I8 alkyl N-methyl glucamide SiUcate: Amoφhous Sodium Silicate (SiO 2 :Na 2 O ratio = 2.0) NaSKS-6: Crystalline layered silicate of formula d-Na 2 Si 2 O 5 Carbonate: Anhydrous sodium carbonate Phosphate: Sodium tripolyphosphate MA/AA: Copolymer of 1:4 maleic/acrylic acid, average molecular weight about 80,000 Polyacrylate: Polyacrylate homopolymer with an average molecular weight of 8,000 sold under the tradename PA30 by BASF Gmbh Zeolite A: Hydrated Sodium Aluminosilicate of formula Naι 2 (AlO 2 SiO 2 ) 12 . 27H 2 O having a primary particle size in the range from 1 to 10 micrometers Citrate: Tri-sodium citrate dihydrate Citric: Citric Acid Perborate: Anhydrous sodium perborate monohydrate bleach, empirical formula NaBO 2 .H 2 O 2 PB4: Anhydrous sodium perborate tetrahydrate Percarbo¬ nate: Anhydrous sodium percarbonate bleach of empirical formula 2Na 2 CO 3 .3H 2 O 2 TAED: Tetraacetyl ethylene diamine CMC: Sodium carboxymethyl cellulose DETPMP: Diethylene triamine penta (methylene phosphonic acid), marketed by Monsanto under the Tradename Dequest 2060 PVP: Polyvinyl pyrroUdone polymer EDDS: Ethylenediamine -N, N'- disuccinic acid, [S,S] isomer in the form of the sodium salt Suds Suppressor: 25% paraffin wax Mpt 50°C, 17% hydrophobic silica, 58% paraffin oil Granular Suds: 12% SUicone/silica, 18% stearyl alcohol, 70% starch in granular form Sulphate: Anhydrous sodium sulphate HMWPEO: High molecular weight polyethylene oxide TAE 25: Tallow alcohol ethoxylate (25) Composition 1 A granular fabric cleaning composition in accordance with the invention may be prepared as follows: Sodium linear C 12 alkyl 6.5 benzene sulfonate Sodium sulfate 15.0 Zeolite A 26.0 Sodium nitrilotriacetate 5.0 Enzyme of the invention 0.1 PVP 0.5 TAED 3.0 Boric acid 4.0 Perborate 18.0 Phenol sulphonate 0.1 Minors Up to 100 Composition 2 A compact granular fabric cleaning composition (density 800/1) in accord with the invention may be prepared as follows: 45AS 8.0 SUBSTITUTE SHEET 25E3S 2.0 25E5 3.0 25E3 3.0 TFAA 2.5 ZeoUte A 17.0 NaSKS-6 12.0 Citric acid 3.0 Carbonate 7.0 MA/AA 5.0 CMC 0.4 Enzyme of the invention 0.1 TAED 6.0 Percarbonate 22.0 EDDS 0.3 Granular suds suppressor 3.5 water/minors Up to 100% Composition 3 Granular fabric cleaning compositions in accordance with the invention which are especially useful in the laundering of coloured fabrics were prepared as follows: LAS 10.7 TAS 2.4 TFAA - 4.0 45AS 3.1 10.0 45E7 4.0 25E3S - 3.0 68E11 1.8 25E5 - 8.0 Citrate 15.0 7.0 Carbonate - 10 Citric acid 2.5 3.0 ZeoUte A 32.1 25.0 Na-SKS-6 - 9.0 MA/AA 5.0 5.0 DETPMP 0.2 0.8 Enzyme of the invention 0.10 0.05 Silicate 2.5 - Sulphate 5.2 3.0 PVP 0.5 „ Poly (4-vinylpyritiine)-N- 0.2 Oxide/copolymer of vinyl- imidazole and vinyl- pyrroUdone Perborate 1.0 Phenol sulfonate 0.2 Water/Minors Up to 100% Composition 4 Granular fabric cleaning compositions in accordance with the invention which provide "Softening through the wash" capability may be prepared as follows: 45AS - 10.0 LAS 7.6 - 68AS 1.3 - 45E7 4.0 - 25E3 - 5.0 Coco-alkyl-dimethyl hydroxy¬ 1.4 1.0 ethyl ammomum chloride Citrate 5.0 3.0 Na-SKS-6 - 11.0 Zeolite A 15.0 15.0 MA/AA 4.0 4.0 DETPMP 0.4 0.4 Perborate 15.0 - Percarbonate - 15.0 TAED 5.0 5.0 Smectite clay 10.0 10.0 HMWPEO - 0.1 Enzyme of the invention 0.10 0.05 Silicate 3.0 5.0 Carbonate 10.0 10.0 Granular suds suppressor 1.0 4.0 CMC 0.2 0.1 Water/Minors Up to 100% Composition 5 Heavy duty liquid fabric cleaning compositions in accordance with the invention may be prepared as foUows: I II LAS acid form - 25.0 Citric acid 5.0 2.0 25AS acid form 8.0 - 25AE2S acid form 3.0 - 25AE7 8.0 - CFAA 5 - DETPMP 1.0 1.0 Fatty acid 8 - Oleic acid - 1.0 Ethanol 4.0 6.0 Propanediol 2.0 6.0 Enzyme of the invention 0.10 0.05 Coco-alkyl dimethyl _ 3.0 SUBSTITUTE SHEET hydroxy ethyl ammonium chloride Smectite clay 5.0 PVP 2.0 Water / Minors Up to 100% SHEET Composition 6 Alcohol ethoxylate C 8 , 5-7 EO 6 4 Alkyl benzene sulfonate; C n .u 5 20 Linear alkyl sulfate; 5 0 (e.g Sulfopon) Soap 1 4 Sodium carbonate 10 0 Zeolith Na-A 25 35 Sodium sillicate; Na 2 O:SiO 2 =3 2 2 Sodium sulfate (Na 2 SO 4 ) 1 20 Polycarboxylate; (Sokalan- CP5) 5 5 Carboxylic acids (Sokalan DCS) 0 4 Sodium perborate tetrahydrate 20 0 TAED 6 0 Suds supressors 5 0 Enzymes including lipolytic enzymes 0.0001 - 0.1 0.0001 - 0.1 (calculated as pure enzyme protein) SUBSTITUTE SHEET Composition 7 A bleach containing detergent composition formulated as a granulate Primary alcohol sulfate (CocoPAS) 5 6 Nonion 3EO (e.g Alkohol ethoylate C12- 5 0 15; 3EO) Nonion 7EO (Alkohol ethoylate C12-15; 8 14 7EO) Sokalan HP22* 0,7 0,8 Soap 1,3 0 Zeolite MAP 39 39 Sodium citrate 4 5 Sodium carbonate 3,3 1 Water/ salts 0,4 0,5 Antifoam/flourescer/perfume 4 5 TAED 5 5 Mn-catalyst 1,7 1,7 Percarbonate 21 21 EDTMP 0,4 0,4 Enzymes including lipolytic enzymes 0.0001 - 0.1 0.0001 - 0.1 (calculated as pure enzyme protein) * a graft polymer as decribed in WO 95/22593 T Composition 8 A non-bleach detergent composition formulated as a granulate Primary alcohol sulfate 6 6 11 9 6 6 (CocoPAS) Nonion 3EO (e.g Alkohol 6 0 4 4 6 0 ethoylate C12-15; 3EO) Nonion 7EO (e.g Alkohol 6 14 6 6 9 15 ethoylate C12-15; 7EO) Sokalan HP22* 0,7 0,7 0,6 0,7 0,5 0,5 Soap 2 2 2 2 2 2 Zeolite MAP 39 39 30 32 40 40 Sodium citrate 25 25 30 32 22 22 Sodium carbonate 1 1 2 2 1 1 Sodium CMC 0 0 0 0 0,7 0,7 Water/ salts 5 6 5 6 6 6 Antifoam/PVP/perfume 3 3 4 4 3 3 EDTMP 1,4 1,4 1,4 1,4 1,4 1,4 Enzymes including lipolytic 0.0001 L - 0.1 enzymes (calculated as pure enzyme protein) * A graft polymer as decribed in the invention WO95622593 SUBSTITUTE SHEET Composition 9 A detergent composition formulated as a granulate Alcohol ethoylat C 12 . 18 , 0,5 0 0 5EO Alkyl glycoside,C 12 . 14 . 5 5 10 Degree of polymerization: 1,4 Linear alkyl sulfate C 16 . 18 10 18 15 (e.g Sulfopon, Henkel) Soap 1 6 6 Sodium carbonate 12 0 0 Zeolith Na-A 23 50 33 Sodium silicate 5 0 3 (SiO 2 :Na 2 O = 3.0) Polycarboxylate (Sokalan CP5) 6 0 0 Carboxylic acids (Sokalan DCS) 0 4 0 Sodium percarbonate 0 0 15 TAED 6 0 5 Sud supressors 6 6 6 Enzymes including lipolytic 0.0001 - 0.1 enzymes (calculated as pure enzyme protein) Water to 100 T Composition 10 A detergent composition formulated as a granulate ZeoUte A 38 38 0 0 0 Sodium disUicate (e.g SKS-6, 0 0 25 30 30 Hoechst) Amoφhes sodium siUcate 5.5 5.5 0 0 0 Na 2 O:SiO 2 = 1:2.0 Sodium carbonate 3 3 3 5 0 Polycarboxylate (Sokalan CP5) 0 2 6 5 5 Mixt. of 80% Alcohol 2.5 2.5 2.5 2.5 2.5 ethoxylate C 8 , 5 EO and 20 % Alcohol ethoxylate C π . 14 , 3 EO (Dehydol LST 80/20) 14 2 2 2 2 2 (e.g Dehydol TA 14) Sodium alkyl benzene 0 0 2 0 0 sulfonate; C, 2 Alkyl sulfate; C ι6 . 18 7.5 7.5 5.5 7.5 7.5 Perborate tetrahydrate 25 25 25 25 25 TAED 2 2 2 2 2 Enzymes including lipolytic 0.0001 - 0.1 enzymes (calculated as pure enzyme protein) Water, perfume, sud supressor to 100 ect. Composition 11 A detergent composition formulated as a granulate Linear alkylbenzene sulfonate 9 Nonionic ethoxylated alcohol 7EO - (e.g 2 Synperonic A7) Nonionic ethoxylated alcohol, 1:1 mixt. of 5 Superionic A3 & A7 (3 / 7 EO groups) Zeolite 4A 29 Poly carboxylate (e.g Sokalan CP 5) 4 Sodium carbonate 7 Granular sodium silicate 4 TAED 8 Sodium perborate monohydrate 16 EDTMP (Ethylene diamine tetramethylene 0.4 phosphonic acid) Nonionic material containing at least 25 0-1 EO groups (e.g Lutensol AT - BASF and BRIJ- ICI) Minor ingrediens (Fluorescer, CMC, salts 4 antifoam granules, perfume Enzymes including lipolytic enzymes 0.0001 - 0.1 (calculated as pure enzyme protein) Moisture to 100 SUBSTITUTE SHEET Composition 12 A detergent composition formulated as a granulate Sodium primary alkyl sulphate (PAS) 6 Nonionic ethoxylated alcohol 3EO - (e.g 7 Synperonic A3) Nonionic ethoxylated alcohol 7EO - (e.g 6 Synperonic A7) Zeolite MAP 36 Stearic acid 2 Tallow 80EO 0.2 Sodium silicate 3 TAED 5 Manganese catalyst 2 Sodium percarbonate 21 Dequest 2047 0,4 Minor ingrediens (Fluorescer, CMC, salts 4 antifoam granules, perfume) Enzymes including Upolytic enzymes 0.0001 - 0.1 (calculated as pure enzyme protein) Moisture to 100 H ET Composition 13 A detergent composition formulated as a granulate DecyUdene or dodecylidene diglycerol 17 0 Decylidene or dodecylidene triglycerol 0 9 C 12-15 EO 7 ethoxylate 9 ZeoUte 32 32 Sodium carbonate 12 12 Alkaline sodium silicate 1 1 Fatty acid soap 2 2 Sodium carboxy methyl cellulose 1 1 Sodium perborate monohydrate 15 15 TAED 7 7 Bleach stabiliser (EDTMP) 0.4 0.4 SiUcone suds suppressor 0.4 0.4 flourescer/perfume 1 1 Enzymes including Upolytic enzymes 0.0001 - 0.1 (calculated as pure enzyme protein) Moisture to 100 SUBSTITUTE SHEET Composition 14 A detergent composition formulated as a granulate Alkyl benzene sulfonate C 10 . 13 18 13 15 11 15 3 Linear alkyl sulfate; C 16 . 18 3 3 4 14 (e.g Sulfopon) Alcohol ethoxylate; C 16 . 18 ; 1.5 0.5 0.5 5 EO Cetyl/oleyl alcohol 5 EO 1.5 1 0.7 1.4 Cetyl/oleyl alcohol 10 EO 1.5 1 0.7 1.4 Alkohol ethoxylate C 14 . 1S ; 7 2 0.5 EO (Dobanol 45-7) Alkohol ethoxylate C 14 . 15 ; 4 6 0.5 EO (Dobanol 45-4) Zeotith Na-A 50 15 42 15 51 42 Na-siUcate 5 5 Na-carbonate 12 12 Na-sulfate 1 45 1 42 Polycarboxylate (Sokalan CP5) 5 3 5 2 5 4 Na-hydrogensulfate 3 Org. acid (Sokalan DCS) 3 1,5 Water 14.5 11.8 10.1 14.5 16.0 13.1 Soap; C 12 , 18 3 3 1 3 4 3 Enzymes including lipolytic 0.0001 - 0.1 enzymes (calculated as pure enzyme protein) Composition 15 A powdered detergent composition Sodium sulfate to balance * As described in US 5,308,530 Composition 16 An aqueous liquid washing agent Alkyl benzene sulfonate; Cιo-ι 3 ; 0 0 0 0 9 17 0 0 0 Monoethanolaminsalt Alkyl ether sulphate, C 4 ; 2 EO 16 10 10 21 11 0 14 38 21 Sodium lauryl sulfate 0 5 5 0 0 0 4 0 0 Soap (C 12 ., 8 - fatty acid) 5 5 5 5 5 5 5 5 5 Alkohol ethoxylate C, 2 . 15 ; 7EO 22 30 30 30 30 30 20 25 30 Alkylglycoside, C 8 . 10 - degree of 15 0 7 0 0 0 0 0 0 oUgomerization: 1,6 Alkylglycoside, C 12 . 16 - degree of 0 5 0 5 5 5 10 0 5 oligomerization: 1,4 1,2-propanediol 15 15 15 15 15 15 15 15 15 Ethanol 5 5 5 5 5 5 5 5 5 Enzymes including lipolytic 0.0001 - 0.1 enzymes (calculated as pure enzyme protein) Water to 100 Composition 17 A Uquid detergent composition Fattyacid monoglyceride (e.g Cutina AGS, 0.5 Henkel) C 12 -ι 8 fatty acid (e.g Edenor K 12-18, 5 Henkel) Alkohol ethoxylate C 12 . 18 ; 7EO 20 Alkylglycoside, C 4 .degree of 20 polymerization: 1,4 Alkyl sulfate (e.g Sulfopon K35, 5 Henkel) Ethanol 5 1,2 propylene glycol 8 Enzymes including lipolytic enzymes 0.0001 - 0.1 (calculated as pure enzyme protein) Water to 100 Composition 18 A detergent composition formulated as non-aqueous Uquid detergent Alkohol ethoxylate C10-12; 7EO 28 Alkohol ethoxylate C10-12; 3EO 23 Glycerol triacetate 6 Silicone antifoam 1.5 Alkyl benzene sulphonic acid 7 Sodium carbonate 20 Calcite 7 Antiseeding polymer 2 SiUca 4 Carboxy methyl cellulose 2 Peracids* 0 - 1 Brightener 0,2 Perfume 0.6 Enzymes including lipolytic enzymes 0.0001 - 0.1 (calculated as pure enzyme protein) * As described in WO 95/06104 SUBSTITUTE SHEET Composition 19 A detergent composition formulated as a non ; aqueous liquid detergent: Decylidene or dodecylidene diglycerol 25 C10-15 EO 7 ethoxylate 25 Sodium carbonate 17 Sodium perborate monohydrate 11 Alkylbenzene sulphonic acid 6 Calcium carbonate 6 SUica (dispersant) 4 Silicone suds suppressor 3 flourescer/antiashing 3 polymer/antiredeposition polymer Enzymes including lipolytic enzymes 0.0001 - 0.1 (calculated as pure enzyme protein) Composition 20 A detergent composition formulated as aqueous liq. detergents Decyl- or dodecylidene triglycerol 25 0 Decyl- or dodecyUdene diglycerol 0 12.5 C 10 -i 5 EO 7, ethoxylate 0 12.5 Fatty acid 4.5 4.5 Potassium hydroxide 10 10 ZeoUte 15 15 Citric acid 8 8 Glycerol 2 2 Borax 1.5 1.5 Polymer 1 1 Silicone oil 0.3 0.3 Perfume 0.5 0.5 Enzymes including Upolytic enzymes 0.0001 - 0.1 (calculated as pure enzyme protein) Water to 100 Composition 21 A Uquid detergent composition comprising Sodium C11-C15 Alkylbenzene 8 17 10 7 sulfonate Alcohol ethoxy sulfate (C12- 12 6 1 14, 60% ethylene oxide by weight) Alcohol Ethoxylate (12-14C 8 7 8 16 8 4 alcohol)ethoxylate Alkylpolyglycoside 16 15 Trisodium citrate 0-15 0-15 0-10 0-20 10 10 Soap 0-10 0-15 5 4 Carboxymethylenoxysuccinate, 10 0-20 trisodium Oxysuccinate - tetrasodium 6 MA/AA/Hydrophobe 5-15 2-20 2-15 1-10 5 2-15 teφolymers * Monoethanolamine 1 2 2 0-4 2 Triethanolamine 2 4 4 Sodium carbonate 1 Borax pentahydrate 3,5 4 4 Glycerol 4 6 5 Propylene glycol 10 10 2 5 Formic acid 1 1 1 Calcium chloride 1 1 1 1 1 Quaternary amines 2 Ethoxylated Amine 1 2 1 Alkyldimethyl Amine Oxide 1,5 Na Xylene Sulfonates 3 6 3 2 3 Ethanol 10 2 8 3 3 Flourescers 0.25 0.2 0.25 0.25 0.2 0.15 Perfume 0.2 0.15 0.1- 0.2 0.25 0.1- 0.3 0.25 Enzymes including Upolytic 0.0001 - 0.1 enzymes (calculated as pure enzyme protein) Sodium sulfate to balance * As described in the invention US 5,308,530 Composition 22 An aqueous liquid detergent composition SUicon antifoam 0.3 Citric acid 8 Glycerol 1 2 • Borax 2 KOH 10 Zeolite 4A 8 Polymer: Deflocculating polymer w. 1.0 chemical structure as polymer All in EP 346,995 QCC 200 : Bentonite clay 8 Oleic acid 5 LAS-acid 17 Synperonic A3 5 Synperonic A7 5 PVP 0.3 Perfume 0.5 Enzymes including Upolytic enzymes 0.0001 - 0.1 (calculated as pure enzyme protein) Water To 100 Dishwashing Composition The dishwashing detergent composition comprises a surfactant which may be anionic, non¬ ionic, cationic, amphoteric or a mixture of these types. The detergent wiU contain 0-90% of non-ionic surfactant such as low- to non-foaming ethoxylated propoxylated straight-chain al¬ cohols. The detergent composition may contain detergent builder salts of inorganic and/or organic types. The detergent buUders may be subdivided into phosphorus-containing and non-phos¬ phorus-containing types. The detergent composition usuaUy contains 1-90% of detergent buUders. Examples of phosphoms-containing inorganic alkaline detergent buUders, when present, in¬ clude the water-soluble salts especiaUy alkaU metal pyrophosphates, orthophosphates, po¬ lyphosphates, and phosphonates. Examples of non-phosphorus-containing inorganic buUders, when present, include water-soluble alkaU metal carbonates, borates and sUicates as weU as the various types of water-insoluble crystalUne or amoφhous alumino silicates of which zeo- Utes are the best-known representatives. Examples of suitable organic buUders include the alkaU metal, ammonium and substituted ammomum, citrates, succinates, malonates, fatty acid sulphonates, carboxymetoxy succinates, ammomum polyacetates, carboxylates, polycarboxylates, aminopolycarboxylates, polyacetyl carboxylates and polyhydroxsulphonates. Other suitable organic buUders include the higher molecular weight polymers and co-polymers known to have buUder properties, for example appropriate polyacryUc acid, polymaleic and polyacryUc/polymaleic acid copolymers and their salts. The dishwashing detergent composition may contain bleaching agents of the chlo- rine/bromine-type or the oxygen-type. Examples of inorganic chlorine/bromine-type bleaches are Uthium, sodium or calcium hypochlorite and hypobromite as weU as chlorinated trisodium phosphate. Examples of organic cMorine/bromine-type bleaches are heterocycUc N-bromo and N-chloro imides such as trichloroisocyanuric, tribromoisocyanuric, dibromoisocyanuric and dichloroisocyanuric acids, and salts thereof with water-solubilizing cations such as potassium and sodium. Hydantoin compounds are also suitable. The oxygen bleaches are preferred, for example in the form of an inorganic persalt, prefera- biy with a bleach precursor or as a peroxy acid compound. Typical examples of suitable per¬ oxy bleach compounds are alkati metal perborates, both tetrahydrates and monohydrates, al¬ kaU metal percarbonates, persilicates and peφhosphates. Preferred activator materials are TAED and glycerol triacetate. The dishwashing detergent composition of the invention may be stabilized using conventional stabilizing agents for the enzyme(s), e.g. a polyol such as e.g.propylene glycol, a sugar or a sugar alcohol, lactic acid, boric acid, or a boric acid derivative, e.g. an aromatic borate ester. The dishwashing detergent composition may also comprise other enzymes, in particular an amylase, a protease and/or a ceUulase. The dishwashing detergent composition of the invention may also contain other conventional detergent ingredients, e.g. deflocculant material, fiUer material, foam depressors, anti-cor¬ rosion agents, soU-suspending agents, sequestering agents, anti-soU redeposition agents, dehy- drating agents, dyes, bactericides, fluorescers, thickeners and perfumes. The first wash Upolytic enzyme of the invention may be incorporated in concentrations con- ventionaUy employed in detergents. It is at present contemplated that, in the detergent compo¬ sition of the invention, the lipolytic enzyme may be added in an amount corresponding to 0.00001-1 mg (calculated as pure enzyme protein) of Upolytic enzyme per Uter of wash tiq- uor. Below, SpecificaUy prefened diswashing compositions are exemplified: 1) POWDER AUTOMAΗC DISHWASHING COMPOSITION Nonionic surfactant 0.4 - 2.5% Sodium metasilicate 0 - 20% Sodium disUicate 3 - 20% Sodium triphosphate 20 - 40% Sodium carbonate 0 - 20% Sodium perborate 2 - 9% Tetraacetyl ethylene diamine (TAED) 1 - 4% Sodium sulphate 5 - 33% Enzymes 0.0001 - 0.1% 2) POWDER AUTOMAΗC DISHWASHING COMPOSITION Nonionic surfactant 1 - 2% (e.g. alcohol ethoxylate) Sodium disUicate 2 - 30% Sodium carbonate 10 - 50% Sodium phosphonate 0 - 5% Trisodium citrate dihydrate 9 - 30% Nitrilotrisodium acetate (NTA) 0 - 20% Sodium perborate monohydrate 5 - 10% Tetraacetyl ethylene diamine (TAED) 1 - 2% Polyacrylate polymer (e.g. maleic acid acryhc acid copolymer) 6 - 25% Enzymes 0.0001 - 0.1% Perfume 0.1 - 0.5% Water 5 - 10 3) POWDER AUTOMAΗC DISHWASHING COMPOSITION Nonionic surfactant 0.5 - 2.0% Sodium disiticate 25 - 40% Sodium citrate 30 - 55% Sodium carbonate 0 - 29% Sodium bicarbonate 0 - 20% Sodium perborate monohydrate 0 - 15% Tetraacetyl ethylene diamine (TAED) 0 - 6% Maleic acid/acryhc 0 - 5% acid copolymer Clay 1 - 3% Polyamino acids 0 - 20% Sodium polyacrylate 0 - 8% Enzymes 0.0001 - 0.1% 4) POWDER AUTOMAΗC DISHWASHING COMPOSITION Nonionic surfactant 1 - 2% ZeoUte MAP 15 - 42% Sodium dis icate 30 - 34% Sodium citrate 0 - 12% Sodium carbonate 0 - 20% Sodium perborate monohydrate 7 - 15% Tetraacetyl ethylene diamine (TAED) 0 - 3% Polymer 0 - 4% Maleic acid/acryhc acid copolymer 0 - 5% Organic phosphonate 0 - 4% Clay 1 - 2% Enzymes 0.0001 - 0.1% Sodium sulphate Balance 5) POWDER AUTOMAΗC DISHWASHING COMPOSITION Nonionic surfactant 1 - 7% Sodium disUicate 18 - 30% Trisodium citrate 10 - 24% Sodium carbonate 12 - 20% Monopersulphate (2 KHSO 5 .KHSO 4 .K 2 SO 4 ) 15 - 21% Bleach stabilizer 0.1 - 2% Maleic acid/acryUc acid copolymer 0 - 6% Diethylene triamine pentaacetate, pentasodium salt 0 - 2.5% Enzymes 0.0001 - 0.1% Sodium sulphate, " water Balance 6) POWDER AND LIQUID DISHWASHING COMPOSITION WITH CLEANING SURFACTANT SYSTEM Nonionic surfactant 0 - 1.5% Octadecyl dimethylamine N-oxide dihydrate 0 - 5% 80:20 wt.C18/C16 blend of octadecyl dimethylamme N-oxide dihydrate and hexadecyldimethyl amine N- oxide dihydrate 0 - 4% 70:30 wt.C18/C16 blend of octadecyl bis (hydroxyethyl)amine N-oxide anhydrous and hexadecyl bis 0 - 5% (hydroxyethyl)amine N-oxide anhydrous Ci 3 -C 15 alkyl ethoxysulfate with an average degree of ethoxylation of 3 0 - 10% C 12 -C, 5 alkyl ethoxysulfate with an average degree of ethoxylation of 3 0 - 5% Cι 3 -C, 5 ethoxylated alcohol with an average degree of ethoxylation of 12 0 - 5% A blend of C 12 -C 1S ethoxylated alcohols with an aver¬ age degree of ethoxylation of 9 0 - 6.5% A blend of Cι 3 -C, 5 ethoxylated alcohols with an aver¬ age degree of ethoxylation of 30 0 - 4% Sodium disUicate 0 - 33% Sodium tripolyphosphate 0 - 46% Sodium citrate 0 - 28% Citric acid 0 - 29% Sodium carbonate 0 - 20% Sodium perborate monohydrate 0 - 11.5% Tetraacetyl ethylene diamine (TAED) 0 - 4% Maleic acid/acryhc acid copolymer 0 - 7.5% Sodium sulphate 0 - 12.5% Enzymes 0.0001 - 0.1% 7) NON-AQUEOUS LIQUID AUTOMAΗC DISHWASHING COMPOSITION Liquid nonionic surfactant (e.g. alcohol ethoxylates) 2.0 - 10.0% AlkaU metal silicate 3.0 - 15.0% AlkaU metal phosphate 20.0 - 40.0% Liquid carrier selected from higher glycols, polyglycols, polyoxides, glycolethers 25.0 - 45.0% Stabilizer (e.g. a partial ester of phosphoric acid and a C 16 -C 18 alkanol) 0.5 ■ - 7.0% Foam suppressor (e.g. sUicone) 0 1.5% Enzymes 0.0001 - 0.1% 8) NON-AQUEOUS LIQUID DISHWASHING COMPOS-TITON Liquid nonionic surfactant (e.g. alcohol ethoxylates) 2.0 - 10.0% Sodium silicate 3.0 - 15.0% AlkaU metal carbonate 7.0 - 20.0% Sodium citrate 0.0 - 1.5% StabiUzing system (e.g. mixtures of finely divided sili¬ cone and low molecular weight dialkyl polyglycol ethers) 0.5 - 7.0% Low molecule weight polyacrylate polymer 5.0 - 15.0% Clay gel thickener (e.g. bentonite) 0.0 - 10.0% Hydroxypropyl ceUulose polymer 0.0 - 0.6% Enzymes 0.0001 - 0.1% Liquid carrier selected from higher lycols, polyglycols, polyoxides and glycol ethers Balance 9) THIXOTROPIC UQUID AUTOMATIC DISHWASHING COMPOSITION Cι 2 -C 14 fatty acid 0 - 0.5% Block co-polymer surfactant 1.5 - 15.0% Sodium citrate 0 - 12% Sodium tripolyphosphate 0 - 15% Sodium carbonate 0 - 8% Aluminium tristearate 0 - 0.1% Sodium cumene sulphonate 0 - 1.7% Polyacrylate thickener 1.32 - 2.5% Sodium polyacrylate 2.4 - 6.0% Boric acid 0 - 4.0% Sodium formate 0 - 0.45% Calcium formate 0 - 0.2% Sodium n-decydiphenyl oxide disulphonate 0 - 4.0% Monoethanol amine (MEA) 0 - 1.86% Sodium hydroxide (50%) 1.9 - 9.3% 1,2-Propanediol 0 - 9.4% Enzymes 0.0001 - 0.1% Suds suppressor, dye, perfumes, water Balance 10) UQUID AUTOMATIC DISHWASHING COMPOSITION Alcohol ethoxylate 0 - 20% Fatty acid ester sulphonate 0 - 30% Sodium dodecyl sulphate 0 - 20% Alkyl polyglycoside 0 - 21% Oleic acid 0 - 10% Sodium disUicate monohydrate 18 - 33% Sodium citrate dihydrate 18 - 33% Sodium stearate 0 - 2.5% Sodium perborate monohydrate 0 - 13% Tetraacetyl ethylene diamine (TAED) 0 - 8% Maleic acid/acryhc acid copolymer 4 - 8% Enzymes 0.0001 - 0.1% 11) LIQUID AUTOMAΗC DISHWASHING COMPOSITION CONTAINING PRO¬ TECTED BLEACH PARTICLES Sodium silicate 5 - 10% Tetrapotassium pyrophosphate 15 - 25% Sodium triphosphate 0 - 2% Potassium carbonate 4 - 8% Protected bleach particles, e.g. chlorine 5 - 10% Polymeric thickener 0.7 - 1.5% Potassium hydroxide 0 - 2% Enzymes 0.0001 - 0.1% Water Balance 11) Automatic dishwashing compositions as described in 1), 2), 3), 4), 6) and 10), wherein perborate is replaced by percarbonate. 12) Automatic dishwashing compositions as described in 1) - 6) which additionaUy contain a manganese catalyst. The manganese catalyst may, e.g., be one of the compounds described in "Efficient manganese catalysts for low-temperature bleaching", Nature 369. 1994, pp. 637- 639. Furthermore, the first wash Upolytic enzyme of the invention may be used in softening com¬ positions: The Upolytic enzyme of the invention may be used in fabric softeners, e.g. as described in Surfactant and Consumer Products, Ed. by J. Falbe, 1987, pp 295-296; Tenside Surfactants Detergents, 30 (1993), 6, pp 394-399; JAOCS, Vol. 61 (1984), 2, pp 367-376; EP 517 762; EP 123 400; WO 92/19714; WO 93/19147; US 5,082,578; EP 494 769; EP 544 493; EP 543 562; US 5,235,082; EP 568 297; EP 570237. MATERIALS AND METHODS Lipase activity (LU) A substrate for Upase was prepared by emulsifying glycerine tributyrate (MERCK) using gum-arabic as emulsifier. Lipase activity is assayed at pH 7 using pH stat method. One unit of Upase activity (LU) is defined as the amount needed to Uberate one micromole fatty acid per minute. Strains and plasmids: Humicola lanuginosa DSM 4109 available from the Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Mascheroderweg lb, D-3300 Braunschweig, Federal RepubUc of Germany (EP 305,216) Saccharomyces cerevisiae YNG318: MATa Dpep4[cir + ] ura3-52, Ieu2-D2, his 4-539 Aspergillus oryzae IFO 4177 A. oryzae A1560-T40, a protease deficient derivative of A. orvzae IFO 4177 (WO 91/17243). A. oryzae JaL 125: Aspergillus oryzae IFO 4177 avaUable from Institute for Fermention, Osaka; 17-25 Juso Hammachi 2-Chome Yodogawa-ku, Osaka, Japan, having the alkaline protease gene named "alp" (described by Murakami K et al., (1991), Agric. Biol. Chem. 55, p. 2807-2811) deleted by a one step gene replacement method (described by G. May in "AppUed Molecular Genetics of FUamentous Fungi" (1992), p. 1-25. Eds. J. R. Kinghorn and G. Turner; Blackie Academic and Professional), using the A. oryzae pyrG gene as marker. Absidia reflexa ATTC 44896 is available from ATCC (American Type Culture CoUec¬ tion, 12301, Parklawn Drive, RockvUle, Maryland 20852. USA) as Absidia reflexa ATTC 44896 and from IFO (Institute for Fermentation, 17-85 Juso-horrmachi 2-chomee, Yodogawa-ku, Osaka 532, Japan) as Absidia reflexa IFO 5874 as described in WO 96/113578 (Novo Nordisk A/S). Yeast cell YPH499 (Stratagene) E.coli DH10B (Gibco) pTiK04: constructed from pJSO37 including the mature Ab reflexa NL 127 lipase gene with a SPIRR encoding extension upstream of the start of the lipase gene. pTiK05: As pTiK04 without the SPIRR extension pTiK06: pTik04 with the MFαl signal sequence pTiK07: pTik05 with the MFαl signal sequence pYESHL is a yeast/E. coli shuttle vector that expresses and secretes a low level of the H. lanuginosa Upolytic enzyme in yeast. More SpecificaUy pYESHL is a derivative of pYES2 in which the GALl promoter was excised and the H. lanuginosa Upolytic enzyme gene and the TPI (triose phosphate isomerase) promoter from S. cerevisiae (Alber, T. and Kawasaki, G., J.Mol.Appl. Genet 1, 419-434 (1982) were cloned between the Sphl and Xbal sites. A restriction map of pYESHL is shown in Fig. 1. PJSO37 (S. cerevisiae expression plasmid)(J.S.Okkels, (1996)"A URA3-promoter deletion in a pYES vector increases the expression level of a fungal Upase in Saccharomyces cerevisiae. Recombinant DNA Biotechnology HI: The Integration of Biological and Engineering Sci¬ ences, vol. 782 of the Annals of the New York Academy of Sciences) More SpecificaUy, the expression plasmid pJSO37, is derived from pYES 2.0 by replacing the inducible GAL1- promoter of pYES 2.0 with the constitutively expressed TPI (triose phosphate isomerase)- promoter from Saccharomyces cerevisiae (Albert and Karwasaki, (1982), J. Mol. Appl Genet., 1, 419-434), and deleting the URA3 promoter. A restriction map of pJSO37 is shown in Fig. 6. pYES 2.0 (Invitrogen Coφ., UK) p960 A. oryzae expression plasmid (described in EP 305 216) pUC19 (Yanish-Perron et al. (1985) Gene 33, 103-119) pHD414 (Aspergillus expression vector being a derivative of the plasmid p775 described in EP 238 023). The construction of pHD414 is further described in WO 93/11249). PJVi245 (See Figure 8) pCaHj383 (see Figure 8) pCaHj ' 385 (see Figure 8) Low calcium filter assay Procedure 1) Provide SC Ura " repUca plates (useful for selecting strains carrying the expression vector) with a first protein binding filter (Nylon membrane) and a second low protein binding filter (CeUulose acetate) ' on the top. 2) Spread yeast ceUs containing a parent Upolytic enzyme gene or a mutated Upolytic enzyme in question on the double filter and incubate for 2 or 3 days at 30°C. 3) Keep the colonies on the top filter by transferring the topfilter to a new plate. 4) Remove the protem binding filter to an empty petri dish. 5) Pour an agarose solution comprismg an oUve oU emulsion (2% P.V.A.:OUve oU=3:l), Brilliant green (indicator,0.004%), 100 mM tris buffer pH9 and EGTA (final concentration 5mM) on the bottom filter so as to identify colonies expressing Upolytic activity in the form of blue-green spots. 6) Identify colonies found in step 5) having a reduced dependency on calcium as compared to the parent Upolytic enzyme. Dobanol®25-7 filter assav: The screening for an improved tolerance towards a detergent component is performed by use of a filter assay conesponding to that described above except for the fact that the solution defined in 5) further comprises 0.02% Dobanol®25-7. An alternative screening assay is the following: 1) Provide SC Ura-plates (useful for selecting strains carrying an expression vector) with first a protein binding filter (e.g. nylon) foUowed by a non-protein bindmg filter (e.g. CeUulose acetate) on the top. 2) Spread yeast ceUs containing a parent Upase gene or a mutated Upase gene on the filter and incubate for 3 or 4 days at 30°C. 3) Keep the colonies on the top filter by transferring the topfilter to a new plate. EET 4) Remove the protein binding filter to a petri dish containing: An agarose solution comprising an oUve oU emulsion (2% P.V.A.:OUve oU=2:l), Brilliant green (indicator,0.004%), 100 mM tris buffer pHIO and the detergent or detergent component, e.g. PCS-plates. The protein binindg filter should have teh colony side facing the screening plate. 5) Identiiy colonies expressing Upase activity in the form of blue-green spots found in step 4) Alternatively, the non-protein binding filter (or a protein binding filter) carrying the yeast colonies may be used directly on the screening plate. Construction of random mutagenized libraries a) Rationale and mathematics behind the desing of random mutagenized libraries The overall rationale for the random mutagenesis is to mimic the evolution in nature where a low continuous mutagenesis is coupled to a continuous selection for a better mu¬ tant which is then further mutagenized. Similarly, the recent in vitro evolution studies described in the Utterature have been performed with consecutive rounds of mutagenesis with increasing selection pressure (for a review see Joyce 1992). We have adapted this by using the wt gene in the first rounds of mutagenesis. Improved variants are then used in the next rounds of mutagenesis (to improve by small steps). We have screened under wash correlated conditions that are only just enough to knock out the wt enzyme activity or improved variants activity. This means that we increase the stringency of screening when better and better variants are isolated. To increase the number of exchanges and to increase the likelyhood of finding im¬ proved variants, localized random mutagenesis have also been performed. Important re¬ gions deduced from the structure of Lipolase and from results from site-directed mutagenesis were selected. E.g. the whole lipid contact zone was considered as important for improvement, especially the lid region and the lid-contact regions. The lipid contact zone corresponds to 7 regions on the gene which have been mutated. Combinations of the regions have also been done. b) Random mutagenesis of an entire lipolytic enzyme coding gene The plasmid pYESHL is treated with 12 M formic acid for 20 min. at room temperature. The resulting Upolytic enzyme encoding gene is ampUfied from the formic acid SUBSTITUTE SHEET treated plasmid usmg PCR under mutagenic conditions (0.5 mM MnCl 2 and 1/5 the normal amount of ATP, see e.g. Leung et al., 1989. This treatment is expected to give a broad range of mutations since formic acid gives mainly transversions and PCR generated mutations mainly transitions. The resulting PCR fragments are cloned either by double recombination (Muhlrad et al., 1992) in vivo into the shuttle vector or digestion and Ugation into the shuttle vector and transformation of E. coli. Eight randomly picked clones have been sequenced and were found to contain 2-3 mutations in average - both transversion and transitions. By use of this method seven Ubraries were made containing from 10,000 to 140,000 clones. c) Localized random mutagenesis A mutagenic primer (oUgonucleotide) is synthesized which conesponds to the part of the DNA sequence to be mutagenized except for the nucleotide(s) conesponding to amino acid codon(s) to be mutagenized. Subsequently, the resulting mutagenic primer is used in a PCR reaction with a suitable opposite primer. The resulting PCR fragment is purified and digested and cloned into the shuttle vector. Alternatively and if necessary, the resulting PCR fragment is used in a second PCR reaction as a primer with a second suitable opposite primer so as to aUow digestion and cloning of the mutagenized region into the shuttle vector. The PCR reactions are performed under normal conditions. When synthesizing the oligonucleotides used for the localized random mutagenesis, cal¬ culation of the doping level is important to estimate the mutagenesis frequency. The fre¬ quency of nucleotide exchanges can be calculated using the Binomial distribution for¬ mula: _Jϋ P(i) = i!(N-i)! x p x (l-p) N_i Where N = the number of doped oligo nucleotides; p = the fraction of none wt nucleo¬ tides; i = number of nucleotide exchanges; P(i) = the probability for the i number of exchanges. It is difficult to calculate the exact number of aa exchanges from the number of nucleotide exchanges, because the third position in a codon for most of the aa can be two or all four nucleotides with out changing the aa. The same is the case for the first or SUBSTITUTE SHEET second position for the three aa with 6 codons. For estimating the number of aa ex¬ changes a Monte-Carlo simulation is more appropriate. For example the program caUed RAMHA performs such a simulation (described in Siderovski and Mak 1993). This pro¬ gram simulates the synthesis of e.g. 10,000 oligonucleotides with the desired doping and calculates the frequency of 0 to n aa exchanges. A doping example The relationship between doping and aa exchanges in a region of 13 codons is (calculated using a Monte Carlo simulation): Percent 0 1 2 3 4 5 6 7 doping mutations level 5% 0.2 0.35 0.27 0.13 0.05 0 0 0 10% 0.04 0.13 0.24 0.25 0.19 0.11 0.05 0 15% 0.005 0.03 0.10 0.20 0.24 0.23 0.13 0.07 The possible number of combinations of aa exchanges for 13 aa can be calculated using the formula: y! 20* N = x!(y-x)! y= number of aa mutagenized x = number of aa exchanges 1 aa exchange in 13 aa = 260 possible exchanges 2 - - - - = 31200 - combinations 3 - - - - = 2.3 x lθ6 - From this follows that when screening e.g 100,000 colonies of a Ubrary doped with 10% in 13 codons giving the distribution shown in the above table wiU mean screening of about 13,000 with one aa exchange (13%). There are, however, only 260 possible one aa exchanges, so a large number of the same one aa exchanges are being screened. A higher doping of e.g. 15% (in the above table) will give fewer one aa exchanges (about 3%), however, the two aa exchanges will also be lowered to a degree (10%) that will not en¬ able screening of the 31200 possible combinations with a screenings capacity around 100.000. FinaUy, the aa exchanges are biased by the origin of the wt amino acid. E.g. it takes only one nucleotide exchange to change Glu to Ala, but three from Glu to Phe. This means that the probability is lower for the aa exchanges that requires 2 or 3 nucleotide exchanges than for those that requires one nucleotide exchange. Therefore we have in some cases allowed more than one aa at positions where we know it is possible. We have always chosen G/C at the third position of the codons with four or six codons. This low- 5 ers the bias of the wt codon and also lowers the likely hood of stop codon (from 4.7% to 3.1 % if completely scrambled). For a calculation of the probability of whether a given pool size contain the most probable and least probable replacement mutants, see Palzlrill et al. 1994. 10 Calculation of population distribution in screening of amplified libraries Another consideration may be taken into account. Most of the Ubraries presented herein are amplified in E. coli before they are transformed into yeast. This means that there is a probability for screening the same amplified clone more than once - see box I. 15 Box I Screening of an amplified random mutagenized Ubrary of e.g. 100,000 different clones: 20 64 % of the library is screened when 100,000 colonies have been screened. 90 % - - - - 230,000 - . 95 % - - - - 300,000 - . This is assuming that all 100,000 clones are ampUfied evenly. 25 The following formula can be used to calculate this: lnfl-P^ N = ln(l-l/D) 30 N is the number of screened clones, P is the fraction of different clones screened and D is the total number of different clones. 35 Anti-termination strategies In order to avoid premature truncated proteins nonsense mutations should be avoided in the codons with a potential to form stop codons. For codons that can be substituted with alternative codons with out the potential to form stop codons, the following strategies can be used: Gly: GGA GG(G,C,T) Leu: TT(A,G) CTN Arg: (A,C)GA (A,C)GG or CG(C,T) Ser: TC(A,G) TC(C,T) or AG(C,T) The following aa can, however, only be specified with codons exhibiting stop-codon po¬ tential: Cys, Glu, Lys, Gin, Tφ, and Tyr. Therefore only the doping can be designed to circumvent the random placement of nucleotides producing stop codons. For example: Glu (similar for Lys and Gin): (90%G/5%C,A) (90%A/3.3%C,G,T) (90%A/3.3%C,G,T). No TAA or TAG = STOP. Tyr (similar for Cys): (90%T/3.3%A,C,G) (90%A/3.3%C,G,T) (90%C/10%T). No TAG or TAA = STOP. Tφ: (90%T/3.3%A,C,G) (90%G/5%C,T) (90%G/5%C,T). No TGA or TAG = STOP. Such a strategy will of course abolish certain a.a. exchanges. Using these strategies the number of premature truncated proteins will be lowered dramatically. In vivo recombination of Humicola lanuginosa lipase variants (Gene shuffling) The DNA sequences of a number of Humicola lanuginosa Upase variants can be in vivo recombined in the same mixture. Vectors are prepared from the Upase variants by Ugation into the yeast expression vector pJSO37. AU vectors are cut open with Nrul. DNA fragment of aU homologous DNA sequences are prepared by PCR amplification using standard methods. The DNA fragments and the opened vectors are mixed and transformed into the yeast Saccharomyces cerevisiae YNG318 by standard methods. The recombination host ceU is cul¬ tivated and screened as described above. Apearing transformants are isolated and tested for improved wash performance using one of the filter assay methods described above. Positive transformants are variants with improved wash performance resulting from gene shuffling of homologous DNA sequences. Site-directed in vitro mutagenesis of Upolytic enzyme gene 5 One approach which may be used for introducing mutations into the Upolytic enzyme gene is described in Nelson & Long, Analytical Biochemistry, 180, 147-151 (1989). It involves the 3-step generation of a PCR (polymerase chain reaction) fragment containing the desired mutation introduced by using a chemicaUy synthesized DNA-strand as one of the primers in the PCR-reactions. The construction of a PCR fragment may be performed in accordance with 10 methods known in the art. From the PCR generated fragment, a DNA fragment carrying the mutation can be isolated by cleavage with restriction enzymes and re-inserted mto the expression plasmid. In figures 4 and 5 the method is further outlined. An alternative method for the construction of variants of a H. lanuginosa Upolytic enzyme involves the use of the commercial kit, Chameleon double-stranded, site-directed mutagenesis 15 kit according to the manufacturer's instructions. The gene encoding the Upolytic enzyme in question is inserted into the plasmid pΗD414. In accordance with the manufacturer's instructions the Seal site of the Ampicillin gene of pHD414 is changed to a Mlul site by use of the foUowing primer: Primer 3: AGAAATCGGGTATCCTTTCAG 20 The pHD414 vector comprising the Upolytic gene in question is then used as a template for DNA polymerase and oUgos 7258 and 7770 the sequences of which are disclosed in the Examples hereinafter. The desired mutation (e.g. in the N-terminal of the Upolytic gene) is introduced into the Upolytic gene in question by addition of an appropriate oUgos comprising the desired mutation. When an N-terminal peptide addition is appUed this may be accom- 25 pUshed by mutating codons of the DNA sequence encoding the pro- or prepro part of the par¬ ent Upolytic enzyme. PCR reactions are performed according to the manufacturer's recomendations. DNA sequencing was performed by using AppUed Biosystems ABI DNA sequence model 30 373A according to the protocol in the ABI Dye Terminator Cycle Sequencing kit. Expression of Humicola lanuginosa lipolytic enzyme in Aspergillus oryzae S Cloning of H. lanuginosa Upolytic enzyme is described in EP 305 216. It also describes expression and characterization of the enzyme in A. oryzae. The expression plasmid used is named p960. The expression plasmid used in this apphcation is identical to p960, except for minor modifications just 3' to the Upase coding region. The modifications were made the foUowing way: p960 was digested with Nrul and BamΗI restriction enzymes. Between these two sites the BamΗI/Nhel fragment from plasmid pBR322, in which the Nhel fragment was fiUed in with Klenow polymerase, was cloned, thereby creating plasmid pAOl (figure 2), which contains unique Ba ΗI and Nhel sites. Between these unique sites BamΗI/Xbal fragments from p960 was cloned to give pAΗL (figure 3). Transformation of AspergiUus oryzae (general procedure) Transformation of Aspergillus orvzae (general procedure. 100 ml of YPD (Sherman et al., (1981), Methods in Yeast Genetics, Cold Spring Harbor Laboratory) are inoculated with spores of A. oryzae and incubated with shaking for about 24 hours. The myceUum is harvested by filtration through miracloth and washed with 200 ml of 0.6 M MgSO 4 . The myceUum is suspended in 15 ml of 1.2 M MgSO 4 , 10 mM NaH 2 PO 4 , pH 5.8. The suspension is cooled on ice and 1 ml of buffer containing 120 mg of Novozym * 234, batch 1687 is added. After 5 min., 1 ml of 12 mg/ml BSA (Sigma type H25) is added and incubation with gentie agitation continued for 1.5-2.5 hours at 37°C until a large number of protoplasts is visible in a sample inspected under the microscope. The suspension is filtered through miracloth, the filtrate transfened to a sterile tube and overlayed with 5 ml of 0.6 M sorbitol, 100 mM Tris-HCl, pH 7.0. Centrifugation is per¬ formed for 15 min. at 1000 g and the protoplasts are coUected from the top of the MgSO 4 cushion. 2 volumes of STC (1.2 M sorbitol, 10 mM Tris-HCl, pH 7.5, 10 mM CaCy are added to the protoplast suspension and the mixture is centrifugated for 5 min. at 1000 g. The protoplast peUet is resuspended in 3 ml of STC and repeUeted. This is repeated. FinaUy, the protoplasts are resuspended in 0.2-1 ml of STC. 100 μl of protoplast suspension are mixed with 5-25 μg of p3SR2 (an A. nidulans amdS gene carrying plasmid described in Hynes et al., Mol. and Cel. Biol., Vol. 3, No. 8, 1430- 1439, Aug. 1983) in 10 μl of STC. The mixture is left at room temperature for 25 min. 0.2 ml of 60% PEG 4000 (BDH 29576), 10 mM CaCl 2 and 10 mM Tris-HCl, pH 7.5 is added and carefuUy mixed (twice) and finaUy 0.85 ml of the same solution are added and carefuUy mixed. The mixture is left at room temperature for 25 min., spun at 2.500 g for 15 min. and the peUet is resuspended in 2 ml of 1.2M sorbitol. After one more sedimentation the proto¬ plasts are spread on minimal plates (Cove, (1966), Biochem. Biophys. Acta 113, 51-56) con- taining 1.0 M sucrose, pH 7.0, 10 mM acetamide as nitrogen source and 20 mM CsCl to in¬ hibit background growth. After incubation for 4-7 days at 37°C spores are picked, suspended in sterile water and spread for single colonies. This procedure is repeated and spores of a -sin¬ gle colony after the second re-isolation are stored as a defined transformant. Transformation of A.orvzae A1560-T40 The plasmid carrying a DNA sequence encoding a variant of the invention is transformed mto Aspergillus orvzae A1560-T40, a protease deficient derivative of A. oryzae IFO 4177, using selection on acetamide by cotransformation with pToC 90 harboring the amdS gene from A. nidulans as a 2.7 kb Xba I fragment (Corrick et a (1987), GENE 53, 63-71) on a pUC 19 vector (Yannisch-Perron et a (1985), GENE 33, 103-119). Transformation is performed as described in EP 238023. Fed batch fermentation Fed batch fermentation is performed in a medium comprising maltodextrin as a carbon source, urea as a nitrogen source and yeast extract. The fed batch fermentation is performed by inoculating a shake flask culture of A. oryzae host ceUs in question into a medium com¬ prising 3.5 % of the carbon source and 0.5 % of the nitrogen source. After 24 hours of culti¬ vation at pH 5.0 and 34°C the continuous supply of additional carbon and nitrogen sources are initiated. The carbon source is kept as the Umiting factor and it is secured that oxygen is present in excess. The fed batch cultivation is continued for 4 days, after which the enzymes can be recovered by centrifiigation, ultrafiltration, clear filtration and germ filtration. Further purification may be done by anionexchange chromatographic methods known in the art. Purification of H. lanuginosa lipolytic enzvme variants expressed in S. cerevisiae The fermentation broth is sterile filtered and ammonium acetate (92 g) is added to the filtrate (1400 ml) to give a 0.8 M solution of ammomum acetate. The solution is added onto a Toy¬ opearl Butyl column (XK 16/10). The column is washed with 0.8 M ammomum acetate and the Upolytic enzyme eluted in H 2 O at a flow rate of 5 ml/min. 10 ml fractions are coUected and Upolytic enzyme containing fractions are pooled according to activity in the standard U- pase titration assay. The Upase containing pool are filtered and the pH is adjusted to pH 8.5 and added onto a Q-Sepharose column (HPQ XK 26/10). The column is washed with 200 ml 0.1 M Tris-HCl, pH 8.5 and the Upolytic enzyme eluted in a Unear gradient of 0 to 0.3 M NaCl in 400 ml of 0.1 M Tris-HCl, pH 8.5 at a flow rate of 5 ml/min. 10 ml fractions are coUected and the Upase containing fractions pooled according to activity in the standard Upase titration assay. Fractions containing Upase activity and absoφtion A280/A260 nm is greater than 1.7 are pooled. Purification of H. lanuginosa lipolytic enzyme variants without peptide addition and expressed A. orvzae Fermentation supematant from the A. oryzae culture is centrifuged and ceU debris dis¬ carded. The supernatant is filtered though a 0.45 μ miUipore filter. Then the is precipitated with 60% saturated ammomum sulphate. The precipitate is dissolved in water and soUd am¬ monium acetate added to a final concentration of 0.8 M. The solution is appUed onto a Butyl Toyopearl column pre-equiUbrated with 0.8 M ammomum acetate. The bound enzyme is eluted with gradient using water and 50% ethanol as eluent. Fractions containing enzyme activity are then pooled and conductance is adjusted to lower than 5 mSi and pH is adjusted to 8.5. The pools containing activity are then appUed onto an anion exchange column (e.g. High- performance Q Separose®) pre-equiUbrated with 25 mM Tris-acetate buffer, pH 8.5. The bound activity is eluted with Unear salt gradient using same buffer and 0.5 M sodium chlo¬ ride. Fractions containing high Upolytic enzyme activity are pooled. Fractions containing U- pase activity and absoφtion A280/A260 nm is greater than 1.7 are pooled. Assay for test of First Wash effect The wash performance of Upolytic enzymes was tested in a one cycle wash trial carried out in a thermostated Terg-O-Tometer (TOM) foUowed by linedrying. The experimental conditions were as foUows: Wash Uquor: 1000 ml per beaker Swatches: 7 cotton swatches (9 x 9 cm) per beaker. Stain: Lard coloured with sudan red (Sigma) (0.75 mg sudan red/g of lard). 50 μl of lard/sudan red heated to 70 °C were appUed to the centre of each swatch. After apphcation of the stain the swatches were heated in an oven for 25 minutes at 75 °C. Stored ovemight at room temperature. Water: 3.2 mM Ca 2+ /Mg 2+ (in a ratio of 5:1) Detergent: 5 g/1 of Detergent Composition A or Detergent B. pH adjusted artificiaUy to 10 by NaOH. Detergent Composition A: 0.300 g/1 of alkyl sulphate (AS; C 14 . 16 ) 0.650 g/1 of alcohol ethoxylate (AEO; C 4 , 6EO) 1.750 g/1 of ZeoUte P 0.020 g/1 of Sokalan CP5 0.050 g/1 " of CMC (carboxy- methyl-ceUulose) Mixed in 3.2 mM Ca 2+ /Mg 2+ (5:1) in Milli-Q water, pH 10.2 Detergent Composition B as Detergent Composition A but additional containing the foUowing bleaching agents: 0.900 g/1 Sodium carbonate peroxyhydrate 0.300 g/1 TAED (tetra-acetyl-ethylene-di-irnine) Concentration of Upolytic enzyme (in Detergent composition A as weU as B): 0 and 1250 or 12500 LU/1 Wash Time: 20 minutes Wash temperature: 30°C Rinse: 15 minutes in mnning tap water Drying: Overnight at room conditions (approx. 20°C, 30-40% RH). Evaluation: The reflectance was measured at 460 nm. Afterwards, the fatty matter was extracted from the swatches with chloroform in a Soxhlet extraction apparatus, distilling off the solvent and determining the amount of fatty matter left on the swatches gravimetricaUy. The amout of fatty material may altematively be determined using thin layer chromato- graphy(TLC)/Flame Ionization Detector (FID)]. The percentage of lard removed is determined as: 1) % removal defined as: [(remaining fat on swatches washed with detergent without Upolytic enzyme) minus (remaining fat on swatches washed with detergent with Upolytic enzyme)] divided by (remaining fat on swatches washed with detergent without Upolytic enzyme) and multipUed by 100%, or 2) delta refectance(dR) defined as: (R(swatches washed in detergent with Upase)-R(swatches washed in detergent without Upase). The reflectance (which may also be termed remission) is measured on an Elrepho 2000 apparatus from Datacolor which illuminates the sample with 2 xenon bUtzlambs and measures the amount of reflected Ught so that entirely white conespond to a 100% reflection and entirely black a 0% reflection. Media and substrates: YPD: 10 g yeast extract, 20 g peptone, H 2 O to 810 ml. Autoclaved, 90 ml 20% glucose (sterile filtered) added. LB-medium: 10 g Bacto-tryptone, 5 g Bacto yeast extract, 10 g NaCl in 1 titre water. FG4 medium: 3% soy meal, 3% maltodextrin, 1 % peptone, pH adjusted to 7.0 with 4 M NaOH SC Ura- plates: 10% lOxBasal salts with out amino acids, 0.5% Casamino acids, 0.02% Threonine, 0.01% Tryptophane, 2% Glucose, 1.5% Agar. lOxBasal salts with out amino ac¬ ids: 60g NaOH, 66.8g Yeast nitrogen base with out amino acids (Difco), and lOOg Succinic acid in 1 Uter water. Iitex Agarose HSB 2000 (CAT NO: F90472) BG-reagent: 4 mg/ml BriUiant Green (BG) dissolved in water Substrate 1: 10 ml OUve oU (Sigma CAT NO. 0-1500) 20 ml 2 % polyvinyl alcohol (PVA) The Substrate is homogenised for 15-20 minutes. PCS detergent 10 g/1: SDS 0.52 g Dobanol 25-3 0.60 g Dobanol 25-1 0.58 g NaBO 3 H 2 O 1.50 g Ad 1 titre 0.1 M Tris buffer (pH 9), and dilute further with the Tris buffer to the double concentration of the desired concentration on the PCS plates. PCS-plates Solution for making PCS plates BriUiant Green (BG-reagent) 10 ml Substrate 1 24 ml PCS detergent 500 ml 2% agarose (in TRIS buffer (pH 9) 500 ml Lipase Substrate (Sigma catalogue no. 800-1) BriUiant Green (Merck, art. No. 1.01310) EXAMPLES EXAMPLE 1 Construction of random lipolytic enzyme variants Random mutagenized Ubraries of the entire H. lanuginosa Upolytic enzyme gene and of amino acids (aa) 91-97 and 206-211 thereof were prepared as described in Materials and Methods above. The amino acid regions 91-97 and 206-211 were chosen for the first round of localized mutagenesis since these regions have been found to be important for wash performance. Region 91-97 is a part of the Ud region of the enzyme and region 206-211 constitutes part of the hydrophobic cleft of the enzyme. One oUgonucleotide was synthesized for each of these regions comprising 93 % of the wUd type nucleotides and 2.33% of each of the other three nucleotides at ammo acid codons wanted to be mutagenized. Where possible without changing the amino acid, the third nucleotide (the wobble base) in codons were synthesized with 50%G/50%C to give a larger likelihood for changes to amino acids with one or two codons. The composition of the mutagenic oUgonucleotide of region 91-97 is shown in Table 1. By use of this oUgonucleotide a calculated mutation frequency of approximately 65-70% is obtained in the Ubrary for one amino acid change having been introduced in the parent Upolytic enzyme. The mutation frequency for two or more amino acid changes having been introduced are less than 35 %. This low mutation frequency is chosen to ensure that the observed amino acid changes in positive clones are involved in improving the enzyme and not just "neutral" changes due to a high mutation frequency. The mutagenic primers were used in a PCR reaction with a suitable opposite primer. The resulting PCR fragments were purified and in the case of region 206-211 digested and cloned into the shuttle vector. In the case of region 91-97 the resulting PCR fragment was used in a second PCR reaction as a primer with a second suitable opposite primer. This step was necessary to be able to digest and clone the mutagenized region into the shuttle vector. Libraries of region 91-97 and of region 206-211 have been prepared containing from 10,000 to 80,000 clones/Ubrary. Most colonies were positive (more than 90%) when checked under conditions where the parent Upase is positive, i.e. exhibits Upase activity. The positive reaction was determined in a filter assay with 2.5 mM Ca (instead of 5 mM EGTA). 450,000 colonies were screened from the different Ubraries using the Dobanol®25-7 and low calcium assays described in Materials and Methods above. 25 low calcium positives from the aa 91-97 Ubrary (Ud-region) and twelve Dobanol® 25-7 positives from the whole gene Ubraries were isolated. Fourteen of the low calcium positives from mutagenesis of aa 91- 97 were sequenced. The three other mutations (in codon 83, 103, 145), outside the mutagenized region, can be explained by PCR misincorporation, although the mutation of S83T is a transversion which is quite unusual for PCR misincorporations. Sequence: 5' 5 C G T 5 C 3' T 7 A A 8 G Bottie5ι93%A;2.33%C;2.33%Gand2.33%T T 8 T T A/C T T 5 C C 7 T T 5 C Bottie6ι93%C;2.33% A; 2.33% G and 2.33% T T 8 ' T T 8 A 6 C/G T 5 6 G Bottie7l93%G;2.33% A; 2.33% C and 2.33% T 5 6 G 7 G A 8 AA A 6 T C Bgttie_8ι93%T;2.33%A;2.33%Cand2.33%G 7 Table 1: Illustration of the constmction of oUgonucleotides (SEQ ID No.4) used for localized random mutagenesis of amino acids 91-97 ofthe H. lanuginosa Upolytic enzyme. The numbers presented in the sequence refer to the bottles the composition of which is apppearing to the right of the sequence. Table 2 Strain Variant number type 5 59 I G91A N94K D96A 60 π S83T N94K D96N 61 π S83T N94K D96N 62 m E87K D96V 63 IV E87K G91A D96V 10 64 π S83T N94K D96N 65 m . E87K D96V 67 V N94K F95L D96H 69 V N94K F95L D96H 71 m E87K D96V 15 72 π S83T N94K D96N Table 2: Strain number refers to the originaUy picked clones cloned into Aspergillus 20 expression vector pAHL. Variant type refers to identical clones, which probably have arisen during ampUfication of the random mutagenized Ubrary. Variant types I and π are active in 0.01 % Dobanol®25-7 whUe the rest are inactive like wtid type. Table 3 5 Strain Variant DNA sequence number type (Amino acid number above the sequence) 82 83 84 85 86 87 88 89 90 91 92 wt GGC TCT CGT TCC ATA GAG AAC TGG ATC GGG AAT 1059 I C 60 II A C 61 II A C 62 III A C 63 IV A C 1564 II A C 65 III A C 67 V C 52/68 wt 53 wt 2069 V C 71 III C 72 II A C 73 VI 25 93 94 95 96 97 98 99 100 -103 -145 Wt CTT AAC TTC GAC TTG AAA GAA ATA -ATT -CAT 59 I G G C 60 II G G A 3061 II G G A 62 III T 63 IV C 64 II G 65 III G 3567 V A C A C 52/68 Wt 53 wt 69 V A C A C 71 III G 1 4072 II G A A 73 VI A Table 3: The wUd type seqence is shown at the toptine. Only nucleotides differing from wt are written at the variant sequences. The base of codon 91 and 93 were doped with 1: 1 of C/T 45 and T/G, respectively. Otherwise the nucleotides at codon 91-97 were doped using 93% wt and 2.33 % of the three other nucleotides. SUBSTITUTE SHEET Results from screening a random mutagenized library of aa 85-99 (the lid region) with a doping based on the results obtained from positives from random mutagenesis of aa 91-97. Construction of the random mutagenized library Background Five different types of strong positive mutants were found in screening of the first U- brary of the Ud region (aa 91-97, see previous example). D96 was changed to A, V, N or H and amino acid change E87K, G91A and N94K were found in two to three independ¬ ent mutants in combination with a change of D96 indicating their importance for inde¬ pendence of calcium of Lipolase. Since these mutations were improved with respect to low calcium/Dobanol activity compared to wt in several assays, they were used as a starting point in a second random mutagenesis of the whole lid region. Localized random mutagenesis. The amino acid region aa 85-99 + 83S/T were random mutagenized as foUows. Doping scheme: S83 - 50%S/50%T; E87 - 93%K 7%X; G91 - 93%A/7%X; N94 - 50%K 50%N; D96 - 100%X; the rest were 93%wt/7%X (the percentages refers to the doping of the codons at the nucleotide level (see the sequence of the oligo). The theoretical percentage of the various amino acid codons resulting from these dopings may be calculated using a state of the art computer program). Where possible without chang¬ ing the amino acid, the third nucleotide (the wobble base) in codons were synthesized with 50% G/50 %C to give a larger likelyhood for changes to amino acids with only one or two codons. The composition of the mutagenic oligonucleotide is shown in SEQ ID NO. oligo 2. (SEQ ID NO 6). The none mutagenized nucleotide region were chosen us¬ ing the Oligo program optimizing for stability and no secondary stmcture. This mutagenesis gives a calculated frequency of approximately 93% changes of the starting point (not including S83, N94 and D96) in the Ubrary. This is a very high muta- tion frequency which should give the chance af major changes of the lid region. The mutagenic primer were used in a PCR reaction with a suitable opposite primer. The resulting PCR fragment was used in a second PCR reaction as a primer with a second suitable opposite primer. This step was necessary to be able to digest and clone the mutagenized region into the yeast expression vector pYESHL. It is important to take the A added to the 3' end of the PCR fragment by Taq polymerase into account when de¬ signing a mutagenic primer for such a two step PCR method. In this way random mutagenized libraries of the region aa 85-99 + 83S/T were pre¬ pared. Screening The low calcium filter assay was used with Dobanol and LAS. Screening of the Ud2 library was made with 5 mM EGTA, 0.01 % Dobanol and 0.006% LAS. Several posi¬ tives were detected and isolated, sequenced, transformed into Aspergillus, purified and tested in wash tests. Sequence and wash results of selected positives UnderUned shows conditions used in the filter assay. IF = improvement factor in 3- cycle wash. 5 mM EGTA.0.01 % Dobanol.0.006% LAS E87K,G91A,L93I,N94K,D96A. IF=1.3 5 mM EGTA .0.02% Dobanol N73D,S85T,E87K,G91A,N94K,D96A. IF =1.1 S83T,E87K,W89G,G91A,N94K,D96V. IF=0.8 E87K,G91A,D96R,I100V. IF=5.2 S83T,E87K,Q249R 2g/l PCS E87K,G91A. IF=5.0 Sequence of Oligo-Ud2 (SEQ ID NO. 6): 5'-C ATT TAT 886 888 655 (C/G)(A/C/G/T)(A/C/G/T) 755 (C/G)88 (A/Q57 588 (C/G)76 (7/8)58 665 788 688 (8/7)58 775 ACG AG(A/T) GCC ACG-3' Flask 5: 93% A; 2,33% C; 2,33% G og 2,33 % T. Flask 6: 93% C; 2,33% A; 2,33% G og 2,33 % T. Flask 7: 93% G; 2,33% A; 2,33% C og 2,33 % T. Flask 8: 93% T; 2,33% A; 2,33% C og 2,33 % G. Local random mutagenesis performed on two regions simultaneously Random mutagenized libraries of aa region 56 to 64 and 81 to 99 + 102 were prepared as described in the Materials and Methods using the two oligo nucleotides 004 and 005 as shown in Table 4 in a PCR reaction. Oligo 004 was synthesized for the aa region 81 to 99 + 102 with 93% wt nucleotides and 2.33% of each of the other 3 nucleotides in each posi¬ tion except for the S83 codon which was doped to give 50%S/50%T (see table 4). For aa with 4 or 6 codons a 50%/50% mixture of G/C or A/C was used for the third base (see ta¬ ble 4). For the third base of the Ile codon a 50%/50% mixture of bottle 7 and 8 was used. D96L was used as starting point in the random mutagenesis since it was found in previous good performing variants. Oligo 005 was synthesized for the aa region 56 to 64 with 93% wt nucleotides and 2.33% of each of the other 3 nucleotides in each position. For the posi¬ tions 56, 57 and 62 a bias of positively charged aa among others were introduced (see table 4). For aa with 4 or 6 codons a 50%/50% mixture of G/C or G/T was used for the third base. In general the PCR reaction may introduce mutations outside the doped region which is an advantage since such mutations may benefit to the property of a variant. The oligo 004 was also used in combination with oligo 006 (see table 4) cloned by a double PCR reaction resulting in library covering region 81 to 99 + 102 and region 248- 257,259, 263-269. The oligo 006 was synthesized for the aa region 248-257,259, 263-269 with 93% wt nucleotides and 2.33% of each of the other 3 nucleotides in each position. For aa with 4 or 6 codons a 50%/50% mixture of G/C or A/C was used for the third base (see table 4). For the third base of the Ile codon a 50%/50% mixture of bottle 7 and 8 was used. The oligo 005 and 006 were also used for constmction of random mutagenized libraries using positives in the lid region as a template. Table 4: OUgo.004 102: T C A Lid3 C T G (antisense) 5'- C G C aa 81-99, G C C C BSTITUTE SHEET G G 5 6 A/C G C C?G 5 6 A A 5 7 7 A 7 8 G A 7 8 A T 5 6 A 7 5 8 A 8 C/G 6 G 6 • 8 7/8 -3' A 8 5 Bottle 5: 93% A; T A/C 8 2,33% C; 2,33% G T 5 C/G og 2,33 % T. T 7 7 Bottle 6: 93% C; A 5 5 2,33% A; 2,33% G T 8 A/C og 2,33 % T. 8 8 6 Bottle 7: 93% G: 8 C/G 7 2,33% A; 2,33% C 6 6 A og 2,33 % T. 8 6 G Bottle 8: 93% T; 8 7/8 A/T 2,33% A; 2,33% C 8 5 C/G og 2,33 % G. 6 8 6 5 6 6 Oligo.005 lid-contact aa 56-64: 5'- G A A A T G C A T G C C C C G TE SHEET A 5/7 8 G G 6/8 6 G G T/G C T 7 T A 7 T G G/T C A 7 3' G 8 A . G/C Bottle 5: A 7 93% A; G 7 2,33% C; G G/C 2,33% G og C 6/7 2,33 % T. G 5/7 Bottle 6: G 6/8 93% C; A 7 2,33% A; T 8 2,33% G og G G/C 2,33 % T. C 5 Bottle 7: A 6 93% G; A G/C 2,33% A; C 2,33% C og G 2,33 % T. T Bottle 8: T 93% T; T 2,33% A; C 2,33% C og T 2,33 % G. C T A C T C G T T T 7/5 7/5 7/5 6/7 Otigo 006 6 A C/G G C-terminal 5 C 7 C aa 248- A/C C 7 C 257,259, 7 A 5/8 -3' 263-269 8 A/C 5 Bottle 5: 93% A: C/G 5 8 2,33% C; 2,33% G 5'- 6 7 7 og 2,33 % T. G 6 G 8 Bottle 6: 93% C; C 5/8 ■ T 8 2,33% A; 2,33% G C 5 G A/C og 2,33 % T. C 8 C/G 7 Bottle 7: 93% G; T A/C 7 7 2,33% A; 2,33% C C 5 6 6 og 2,33 % T. T 7 A/C 8 Bottle 8: 93% T; A C/G 7 7 2,33% A; 2,33% C G 6 7 7 og 2,33 % G. A 6 7/8 8 C G 5 8 T A 8 A A A 5 T A/C G 8 T 5 T 6 7 5 Some of the positives obtained from screening these libraries on detergent containing plates are shown below: E56R+D57L+I90F+D96L+E99K E56R+D57L+V60M+D62N+S83T+D96P+D102E D57G+N94K+D96L+L97M E87K+G91A+D96R+I100V+E129K-I-K237M+I252L+P256T+G263A-I-L264Q E56R+D57G+S58F+D62C+T64R+E87G+G91A+F95L+D96P+K98I A47V+D62G+D96L+A121T E56G+D57G+V60E+D62G+N94K+D96L The following variants were obtained from random mutagenesis of the whole gene alone (by PCR or PCR + formic acid as described in the Materials and Methods section) and screened on detergent containing plates: I34V+S54P+F80L+S85T+D96G+R108W+G109V+D111G+S116P+L124S+V132 M+V140Q+ V141A+F142S+H145R+N162T+I166V+F181P+F183S+R205G+A243T+D254G-I- F262L A19T+D167G+E210V+W221L (random mutagenesis based on D167G+E210V) A49P+D167G+E210V (random mutagenesis based on D167G+E210V) EXAMPLE 2 Construction of first wash variants of the H. lanuginosa lipolytic enzyme I. Domain shuffling by recombination and screening 20 H. lanuginosa Upolytic enzyme variants having a very good washing performance (as evaluated in various wash related tests) some of which were constmcted according to Example 1 were aUowed to recombine by an in vivo recombination method in S. cerevisiae YNG318 as described in the Materials and Methods section herein. The Upolytic enzyme variants used are apparent from table 1. Most of these variants had been constmcted by random or localized T random mutagenesis as described in the Materials and Methods section above and screening for a decreased dependence on calcium and an improved tolerance towards the detergent component Dobanol 25-7 (cf. the Materials and Methods section above). Some of the variants are the result of two or more consecutive rounds of mutagenesis and screening. The restriction enzyme opened vector and the PCR fragments apparent from the table below and further discussed in the Materials and Methods section were mixed in a molar ratio of approximately 1:1 and used for transformation of competent S. cerevisiae ceUs (made by the tithium acetate method as described in Current Protocols in Molecular Biology, eds. F.M. Ausubel et al., chapter 13.7, John WUey & Sons, Inc., USA.). The transformed ceUs were plated on filters and screened for a reduced calcium dependency and an increased detergent tolerance using the filter assay described in the Materials and Methods section above. Colonies giving a positive signal were streaked out to single colonies on new plates and filters and re-screened. After 2 to 4 rescreenings positive colonies were fermented as foUows: Fermentation of H. lanuginosa lipolytic enzyme and variants in yeast 10 ml of SC-ura " medium are inoculated with a S. cerevisiae colony and grown at 30°C for 2 days. The 10 ml is used for inoculating 300 ml SC-ura " medium which after inoculation is grown at 30°C for 3 days. The 300 ml is used for inoculation 5 1 of the foUowing G- substrate: 400 g Amicase 6.7 g yeast extract (Difco) 12.5 g L-Leucine (Fluka) 6.7 g (NΗ^SO, 10 g MgSO 4 -7H 2 O 17 g K 2 SO 4 10 ml Trace compounds, cf below 5 ml Vitamin solution, cf below 6.7ml H 3 PO 4 25 ml 20% Pluronic (antifoam) In a total volume of 5000 ml. Tracecompounds: 6.8 g ZnCl 2 54.0 g FeCl 2 6H 2 O 19.1 g MnCl 2 4H 2 O 2.2 g CuSO 4 5H 2 O 2.58 g CoCl 2 0.62 g H 3 BO 3 100 ml HCl (concentrated) In a total volume of 1 1. Vitamin solution: 250 mg Biotin 3 g Thiamine 10 g D-Calciumpantothenat 100 g Myo-Inositol 50 g Chotinechlorid 1.6 g Pyridoxin 1.2 g Niacinamid 0.4 g FoUcacid 0.4 g Riboflavin In a total volume of 1 1. The yeast ceUs were fermented for 5 days at 30°C in 5000 ml medium. Glucose was fed to the fermenter with a start dosage of 100 ml 70% glucose and addition of 400 ml 70% glucose/day. The pH of the fermentation broth was kept at pH 5.0 by controUed addition of a 10% NH 3 solution. Agitation was 300 rpm for the first 22 h foUowed by 900rpm for the rest of the fermentation. Air was given with 1 1 air/1/min for the first 22 h foUowed by 1.5 1 air/1 min for the rest of the fermentation. After purification the capability of the variant in removing lard was tested in the one cycle wash assay described in the Materials and Methods section above. The results were given in Example 2 and 3 hereinbelow. UBSTITUTE SHEET Table 1: Variants used for recombination Humicola lanusinosa lipase variants used for preparing vector (Opened with NmP for invo recombination Cgene shuffling.: E56R,D57L,I90F,D96L,E99K E56R,D57L,V60M,D62N,S83T,D96P,D102E D57G,N94K,D96L,L97M E87K,G91A,D96R,I100V,E129K,K237M,I252L,P256T,G263A,L264Q E56R,D57G,S58F,D62C,T64R,E87G,G91A,F95L,D96P,K98I,(K237M) E210K Humicola lanuginosa lipase variants used for preparing DNA fragments (bv standard PCR amplification of the whole gene from the plasmids containing the variant, for invo recombination fgene shuffling.: S83T,N94K,D96N E87K,D96V N94K,D96A E87K,G91A,D96A D167G,E210V S83T,G91A,Q249R E87K,G91A S83T,E87K,G91A,N94K,D96N,D111N. N73D,E87K,G91A,N94I,D96G. L67P,I76V,S83T,E87N,I90N,G91A,D96A,K98R. E210K S83T,E87K,G91A,N92H,N94K,D96M S85P,E87K,G91A,D96L,L97V. E87K,I90N,G91A,N94S,D96N,I100T. ( TITUTE SHEET I34V,S54P,F80L,S85T,D96G,R108W,G109V,D111G,S116P,L124S,V132M,V140Q,V141 A,F142S,H145R,N162T,I166V,F181P,F183S,R205G,A243T,D254G,F262L. E56R,D57L,I90F,D96L,E99K E56R,D57L,V60M,D62N,S83T,D96P,D102E D57G,N94K,D96L,L97M E87K,G91A,D96R,I100V,E129K,K237M,I252L,P256T,G263A,L264Q E56R,D57G,S58F,D62C,T64R,E87G,G91A,F95L,D96P,K98I,(K237M) 2. Domaine shuffling by traditional cloning of two positives together The Aspergillus expression vector pHD414 containing a DNA sequence encoding the Humicola lanuginosa Upase variant (D57G+N94K+D96L+L97M) were digested with the restriction enzymes Narl and Xbal resulting in two fragments. The fragments were separated by agarose gel electrophoresis and the largest fragment were isolated from the agarose gel. This fragment were tigated to the smaUest fragment from digestion of the variant (S83T+G91A+Q249R) with Narl and Xbal. The Ugation was transformed into E. coli and the resulting plasmid constmctions were isolated from one of the transformants and sequenced to test for the correct assembly. The plasmid was transformed into Aspergillus oryzae fermented and purified as described in the Materials and Methods section.This variant contained the foUowing mutations D57G+N94K+D96L+L97M+Q249R. EXAMPLE 3 Constmction and expression of H. lanuginosa lipolytic enzyme variant HL9 in Aspergillus orvzae JaL125 The variant HL9 contains the following mutations in the mature part: E1P+D57G+N94K+D96L+Q249R and the N-terminal peptide addition SPIRPR fused to EIP (resulting in the overall N-terminal peptide addition SPIRPRP. An N-terminal peptide addition was applied to the parent H. lanuginosa (DSM 4109) lipolytic enzyme having the amino acid and DNA sequence, respectively, apparent from EP 305 216 and in addition carrying the following mutations D57G+N94K+D96L+Q249R in its mature part (inserted by conventional site-directed mutagenesis) in the DNA sequence (EP 305 216). The peptide addition SPIRPRP was applied to the N-terminus of the parent enzymes as follows: Constmction of pIVI220: The plasmid was constmcted using the Chamelon double stranded, site-directed mutagenesis kit from Stratagene according to the described protocol. pΗL296 was used as the plasmid template. Said plasmid contains the gene encoding the H. lanuginosa lipolytic enzyme with the above mentioned mutations (D57G+N94K+D96L+L97M+Q249R) cloned into pHD414. Primer no. 7258 was used as the selection primer. 7258: 5'p gaa tga ctt ggt tga cgc gtc ace agt cac 3 1 (Thus changing the Seal site found in the ampicilUn resistance gene and used for cutting to a Mlul site). Primer no. 7770 was used as the selection primer. 5 7770: 5'p tct age cca gaa tac tgg atc aaa tc 3 ' (Changes the Seal site found in the H. lanuginosa lipase gene without changing the amino acid sequence). Primer no.8479 was used as the mutagenic primer. 8479: 5'p gcg tgg acg gcc ttg get age cct att cgt cct cga ccg gtc teg eag gat ctg (replacing the propeptide and the N-terminal El of the parent H. lanuginosa enzyme (SPIRRE by 0 SPIRPRP). Constmction of pIVI245: The plasmid was constmcted using the Chameleon double-stranded, site-directed mutagenesis kit from Stratagene (cat no. 200509) according to the described protocol. 5 pIVI220 was used as the plasmid templated and primer no.7887 as the selection primer (changing the introduced Mlul site found in the ampicillin resistance gene and used for cutting to a Seal site). 7887: 5'p-gaa tga ctt ggt tga gta etc ace agt cac 3" Primer no. 8932 was used as the mutagenic primer (8932: 5'p-g aac tgg ata gga aat ttg aag ttc ctg ttg aaa gaa ata aat gac 3' (thus changing M97 back to L97 as wildtype 0 and still preserving the two mutations N94K and D96L)). 2. Constmction of the A. orvzae expression plasmid pCaΗi483 pCaHj483 is depicted in Figure 8. It is build from the following fragments : 5 a) The vector pToC65 (WO91/17243) cut with EcoRI and Xbal. b) A 2.7 kb Xbal fragment from A. nidulans carrying the amdS gene (C. M. Corrick et al., (1987), Gene 53, p. 63-71). The amdS gene is used as a selective marker in fungal transformations. The amdS gene has been modified so that the BamHI site normally present in the gene is destroyed. This has been done by introducing a silent point 0 mutation using Primer 3: AGAAATCGGGTATCCTTTCAG (SEQ ID No. 1) c) A 0.6 kb EcoRIIBamHI fragment carrying the A. niger NA2 promoter fused to a 60bp DNA fragment of the sequence encoding the 5 ' untranslated end of the mRNA of the A. nidulans tpi gene. The NA2 promoter was isolated from the plasmid pNA2 (EP 383 779) and fused to the 60 bp φi sequence by PCR. The primer (Primer 4 SEQ ID No. 7) encoding the 60 bp φi sequence had the following sequence : 5'- GCTCCTCATGGTGGATCCCCAGTTGTGTATATAGAGGATTGAGGAAGGAAGAG AAGTGTGGATAGAGGTAAATTGAGTTGGAAACTCCAAGCATGGCATCCTTGC - 3 ' d) A 675 bp Xbal fragment carrying the A. niger glucoamylase transcription terminator. The fragment was isolated from the plasmid pICAMG/Term (EP 238 023). The BamHI site of fragment c) was connected to the Xbal site in front of the transcription terminator on fragment d) via the pIC19R linker (BamHI to Xbal) Constmction of the HL9 expression plasmid pCaHi485 The plasmid pJVi 245 was digested with BamH I and Sal I, and the resulting 904 bp fragment encoding the HL9 lipolytic enzyme was isolated. pCaHj 483 was digested with BamH I and Sal I, and the large vector fragment (6757) was Ugated to the HL9 fragment. The Ugation mixture was used to transform E. coli DH5α cells, and a transformant harbouring the expected plasmid was isolated. The plasmid was termed pCaHj485. 3. Transformation of pCaHi 485 into JaL125 Aspergillus oryzae JaL 125 was transformed with pCaHj 485 using selection on acetamide as described in patent EP 0 531 372. Transformants were spore reisolated twice. Spores from second reisolation of each transformant were used to inoculate 200 μl YPM (1 % yeast extract, 2% peptone, 2% maltose) in 96 well microtiter dishes. The YPM cultures were grown for 4 days at 34 °C, and the higest producers were selected using a p-nitro phenylbutyrate assay: Stock solution: 18 μl p nitro phenyl butyrate was dissolved in 1 ml isopropanol. Working solution: 0.1 ml stock solution was mixed with 10 ml 50 mM Tris/HCl pH 7.5; 10 mM CaCl 2 . Assay: 1 μl of YPM supematant was mixed with 200 μl of working solution in 96 weU microtiterdishes-l- and the color development was measured at 450 nm using an ELISA reader. One transformant was selected for tank fermentation. 4. Tank fermentation of JaL 125/ρCaHi 485 The fermentation was carried out as a fed-batch fermentation using a constant medium temperature of 34°C and a start volume of 1.2 litre. The initial pH of the medium was set to 6.5. Once the pH had increased to 7.0 this value was maintained through addition of 10% H 3 PO 4 . The level of dissolved oxygen in the medium was controlled by varying the agitation rate and using a fixed aeration rate of 1.0 litre air per litre medium per minute. The feed addition rate was maintained at a constant level during the entire fed-batch phase. The batch medium contained maltose symp as carbon source, urea and yeast extract as nitrogen source and a mixture of trace metals and salts. The feed added continuously during the fed-batch phase contained maltose symp as carbon source whereas yeast extract and urea were added in order to assure a sufficient supply of nitrogen. 5. Purification of the first wash lipolytic enzyme 1) Fermentation supematant was filtered through milipore filter Cat. No. AP2504700 FUter type AP25. 2) Fermentation supematant was filtered once more on through the sterile filter from MiUipore membrane Type GS 0.22 micron. 3) Fermenation supematent was then adjusted to 0.8 M ammonium acetate by adding soUd ammonium acetate. 4) A Hydrophobic chromatography on TSK gel Butyl-Toyopearl 650. 50 ml column was packed with the Butyl-Toyopearl matrix. The column was washed and equilibrated with 0.8 M ammonium acetate. One Uter ferementation supematent adjusted with amonium acetate was then appUed on the Butyl column. The column was washed with 0.8 M ammomum acetate till aU unbound material was washed out. Bound material was then eluted with water and 50 % ethanol sequentiaUy. Fractions were coUected and analyzed for Upase activity using Standard LU assay. Fractions containing Upase activity were pooled and dtiuted to adjust conductivity of the pool below 4 mSi and pH to 8.5. 5) Anion exchange chromatography on High Performance Q sepharose (Pharmacia, Code No.17-1014-01). 50 ml column was packed and washed with 50 mM Borate buffer pH 8.5. Pool containing Upolytic enzyme activity was then appUied on The High performance Q se¬ pharose column. Unbound material was washed with the Borate buffer pH 8.5. Bound activity was then eluted with Unear gradient using Borate buffer conatining 1 M Sodium Chloride pH 8.5. Fractions were coUected and assayed for Lipase activity. Fractions containing Lipase activity with a ratio of UV absorbence at A280and A260 more than 1.7 are pooled. EXAMPLE 4 Constmction of N-terminal additions bv random mutagenesis Random mutagenesis of the part of the DNA sequence encoding the N-terminal addition SPIRPRP added to the first amino acid residue of the mature H. lanuginosa Upolytic enzyme (obtainable from DSM 4109) and containing the foUowing further mutations in its mature part: D57G+N94K+D96L+L97M+Q249R was performed. The mutations in the mature part of the parent Upolytic enzyme was performed by PCR driven site-directed mutagenesis using the appropriate primer sequences using the procedures described in WO 95/26215. The peptide addition SPIRPRP was appUed as described in Example 3, (i.e. the last P replacing El). The nucleotide doping scheme of the SPIRPRP codons was as foUows: Oligo 1: 5'- GCG TGG ACG GCC TTG GCC 86.T/A. 66ΪA/T 58(T/A . 67H7A. 66CT/A. 575 66(T/A . GAG GTC TCG CAG GAT CTG -3' (57-mer) (SEQ ID NO 11) the numbers referring to which of the foUowing flasks to be used. Flask 5: 80% A; 6.66% C; 6.66% G og 6.66 % T. Flask 6: 80% C; 6.66% A; 6.66% G og 6.66 % T. Flask 7: 80% G; 6.66% A; 6.66% C og 6.66 % T. Flask 8: 80% T; 6.66% A; 6.66% C og 6.66 % G. A two step PCR reaction protocol was used: The first step with the above primer as the 5' primer and with the primer 2056 (5'gca cgt aat gtt tgt ace 3') as the 3' primer con- ducted using pHL296 as the plasmid template. The product of the first PCR round was used in a new PCR with 4699 (5 'egg tac ccg ggg atc cac 3') as the 5' primer (to intro¬ duce the BamHI site and the first part of the coding sequence) and with the PCR product ET as the 3' primer using the same template. The resulting product was purified on Spin 100 (from Clonetech Lab., Inc.) and cut with BamHI and Pvull. The resulting DNA fragment was purified from the agarose gel with SpinX (Costar) and ligated into the yeast expres¬ sion vector pJSO37 containing the H. lanuginosa Upolytic enzyme gene from pHL296 cloed as a BamHI-Xbal fragment cut with BamHI and Pvull. The resulting DNA was electrotransformed into DH10/DH12 E. coli ceUs (Gibco/BRG Lifetechnologies) using the conventional technique. After transformation into E. coli and amplification the plasmid was purified and trans¬ formed into S. cerevisiae YNG 318. The resulting S. cerevisiae ceUs were screened for good performers in the altemative Upase filter assay containing detergent (3 g/1 of PCS). The posi¬ tives were sequenced and found to contain the foUowing peptide additions: GPIRPRP, SHSRHNA, TAIRPRK, SALRRRP, STRRPRP, SPRRPRT, SPIPPGP, LPFRQRP, SPFRPKL, and SALRRP (termed HLlOsl-10, respectively). The one-cycle wash performance of each of HL10sl-6 was tested as described in the Mate- rials and methods section above (Assay for test of first wash effect) at a temperature 30°C and using 5 g/1 of enzyme inactivated Ariel Futur as detergent. The amount of fatty material removed by each of the modifed enzymes are shown below: Lipase variant Low dosage % lard High dosage % lard removed removed HLvlOsl 1250 LU/1 26 12500 LU/1 54 HLvl0s2 1250 LU/1 22 12500 LU/1 53 HLvl0s3 1250 LU/1 34 12500 LU/1 55 HLvl0s4 1250 LU/1 33 12500 LU/1 55 HLvl0s5 1250 LU/1 23 12500 LU/1 47 HLvlOsό 1250 LU/1 30 12500 LU/1 53 The tendency was that the best performers had more positive charged amino acids in the N- terminal addition. Analogously, random mutagenesis of the N-terminal addition RPRPRPRP added to the H. lanuginosa Upase variant El*+ D57G+ N94K+ D96L+ L97M+ Q249R plus other vari- ants were performed. The nucleotide doping scheme of the RPRPRPRP codons was as fol¬ lows: OUgo 2: 5'-GTC TCT GCG TGG ACG GCC TTG GCG GCG CCA CCT CCA 67(T/A) 66(T/A) 575 66(T/A) 67(T/A) 66(T/A) 575 66(T/A) (6/7) (7/8) (C/G) 57(C/G) C57 (5/7)5(C/G) CTG TTT AAC CAG TTC AAT CTC-3' (93-mer) Flask 5: 80% A; 6.66% C; 6.66% G og 6.66 % T. Flask 6: 80% C; 6.66% A; 6.66% G og 6.66 % T. Flask 7: 80% G; 6.66% A; 6.66% C og 6.66 % T. Flask 8: 80% T; 6.66% A; 6.66% C og 6.66 % G. APPP is added in the N-terminal of the randomly mutagenized RPRPRPRP and prior to the signal peptide in order to protect against proteolytic degradation of the N-terminal addi¬ tion. This may not be required. El was deleted in order to remove one negatively charged amino acid. The amino acids in position 2 to 5 of the mature H. lanuginosa Upase sequence were also mutagenized in order to find improved mutants in this non-stmctural part of the U- pase. Otherwise the procedure is as stated above for the random mutagenesis of SPIRPRP. The following N-terminal peptide additions were obtained: Ala-Pro-Pro-Pro-Arg-Pro-Arg-Leu-Leu-Pro-Ile-Ser( APPPRPRLLPIS) (in addition to the deleted El residue this variant carries the additional mutation D5E in its non-stmctural N- teπninal part of the mature enzyme). Ala-Pro-Pro-Pro-Thr-Arg-Gln-Arg-Gln-Ser-Pro(APPPTRQRQSP) (in addtion to the de¬ leted El residue this variant carries the additional mutations V2L, S3T and D5V in its non-stmctural N-terminal part of the mature enzyme). Ala-Pro-Pro-Pro-Arg-Thr-Ile-Pro-Arg-Ser-Ser-Pro( APPPRTIPRSSP) (in addition to the deleted El residue this variant carries the additional mutations V2L, S3R and D5E in its non-stmctural N-terminal part of the mature enzyme). Ala-Pro-Pro-Pro-Arg-Pro-Arg-Pro-Arg-Pro-Arg-Pro (APPPRPRPRPRP) (in addtion to the deleted El residue this variant carries the additional mutations V2G and D5E in its non- structural N-terminal part of the mature enzyme). Ala-Pro-Pro-Pro-Arg-Thr-Arg-Pro-Arg-Pro-Arg-Ser (APPPRTRPRPRS) (in addtion to the deleted El residue this variant carries the additional mutations V2GL, S3T, Q4P and D5E in its non-stmctural N-terminal part of the mature enzyme). Ala-Pro-Pro-Pro-Lys-Ala-Ser-Pro-Arg-Gln-Arg-Pro (APPPKASPRQRP) (in addtion to the deleted El residue this variant carries the additional mutations V2GL, D5Q and L6M in its non-stmctural N-terminal part of the mature enzyme). EXAMPLE 5 First wash activity of lipolytic enzymes of the invention The first wash activity of lipolytic enzymes was tested using the "Assay for test of First Wash effect" described in the Materials and Methods section above with Detergent Composition A or B. A few of the new first wash lipase are compared to what is considered as being the present state of art within lipolytic enzymes for detergents. Note: π are produced in Aspergillus oryzae as described in example 2. * are produced in yeast as described in example 4. Detergent Composition A Lipolytic Enzyme % removal at % removal at 1250 LU/1 12500 LU/1 E1SPIRPRP+D57G+N94K+D96L+L97M+ 15% 49% Q249R Lumafast™ (Ps. mendocina) 0% 2% Lipomax™ (Ps. pseuodoalcaUgenes L21M) 0% 9% Fusarium solani pisi 0% 0% Lipolase 0% 0% Lipolase Ultra 0% 0% Detergent Composition B Lipolytic Enzyme % removal at % removal at 1250 LU/1 12500 LU/1 E1SPIRPRP+D57G+N94K+D96L+L97M+ 15% 46% Q249R Lumafast™ (Ps. mendocina) 6% 6% Lipomax™ (Ps. pseuodoalcaligenes L21M) 0% 0% Liposam™ 4% 7% Fusarium solani pisi 2% 5% Lipolase 5% 6% Lipolase Ultra 6% 0% Additional examples: Detergent Composition A Lipolytic Enzyme % removal at 12500 LU/1 SPIRR+D57G+G59V+N94K+D96L+L97M+S116P+S1 42% 70P+Q249R * SPIRR+A49P+D167G+E210V * 44% SPIRR+E56K+D57G-I-D62R-I-S83T+S85F+D96L+D10 36% 2Y+E210K * SPIRR+N94K-I-F95L+D96H-I-N101S+F181L-I-D234Y+I 41 % 252L+P256T+G263A+L264Q * EXAMPLE 6 Activity-in-Detergent (AiD) assay The AiD assay is an analytical assay that is useful for selecting parent Upolytic enzymes to be used in the construction of a first wash lipolytic enzyme as described herein. Equipment: Water bath with 150 ml beakers. Stirring is obtained by an agitator. Lipolytic enzyme dosage: 12500 LU/1. Substrate:6 pieces (3.5*3.5 cm) of cotton with 6 μl olive oil for one test. Detergent:0.5 g/1 model Uquid detergent * dissolved in 0.36 mM Ca 2 7Mg 2- (5:1), adjusted to pH 10, 100 ml per beaker. After stirring the sample for 60 min. at 30°C the remaining detergent on the swatches is removed by addition of tap water for 15 min. The swatches are put into a flask containing 10 ml Tetrahydrofuran and 6.25 ml 4 M HCl and evaporated over night, after which the sample is redissolved in Tetrahydrofuran. The fat composition is determined by TLC/FID and the amount of % FFA (free fatty acids) is used to distinguish between the Upolytic enzymes. * detergent formulation below Note π are produced in Aspergillus oryzae ast as described in example 2 * all variants are produced in yeast as described in example 4. AiD assay Lipolytic Enzyme % FFA SP-tRR+D57G+G59V+N94K+D96L+L97M+S116P+ 20% S170P+Q249R * SPIRR+A49P+D167G+E210V * 25% SPffiR+E56K+D57G+D62R+S83T-l-S85F+D96L+ 25% D102Y+E210K * SP-rRR+N94K+F95L+D96H+N101S+F181L-l-D234Y+I 20% 252L+P256T+G263A+L264Q * ElSPIRPRP+D57G+N94K+D96L+L97M+Q249Rπ 27% Lumafast™ (Ps. mendocina) 5% Lipomax™Cos (Ps. pseudoalcaUgenes) 31% Fusarium solaiή pisi 6% Lipolase 5% Lipolase Ultra 5% TITUTE SHEET Model liquid detergent: Component % w/w LAS 17.50 AEO 14.40 DTSA 10.00 Oleic acid 3.00 Coconut oil 5.00 MEA 14.50 MPG 10.70 Ethanol 1.40 Phosphonate 1.00 Boric acid 0.80 Citric acid 3.90 Sodium chloride 0.13 Potassium chloride 0.38 Hydrochloric acid 4 6.00 M Water 9.7 EXAMPLE 7 The first wash activity of a large number of potential first wash Upolytic enzyme was tested using the "Assay for test of First Wash effect" described in the Materials and Methods section above with a specific commercial detergent - Ariel Futur (commerciaUy avaUable batch No.4279 B 23:35). The enzymes already present in the detergents were inactivated by heat (4 minutes at 85°C in micro oven) prior to wash. The first table can be used to compare to example 5 & 6. Afterwards the results are divided as foUows: a) % removal when dosing after LU-units (see methods & materials for definition) b) % removal when dosing after miUigrams of pure enzyme protein c) delta Reflectance when dosing after LU-units (see methods & materials for definition) d) delta Reflectance when dosing after miUigrams of pure enzyme protein Note: π are produced in Aspergillus oryzae as described in example 2 * are produced in yeast as described in example 4. The following results were obtained: Enzyme Inactivated Commercial European Detergent Lipolytic Enzyme % removal at 1250 LU/1 12500 LU/1 SPIRR+D57G+G59V+N94K+D96L+L97M+ 12% 37% S116P+S170P+Q249R * SPIRR+A49P-I-D167G+E210V * 8% 38% SPIRR+E56K+D57G+D62R+S83T+S85F+ 8% 34% D96L+D102Y+E210K * SPIRR+N94K+F95L+D96H+N101S+F181L+ 11 % 37% D234Y+I252L+P256T+G263A+L264Q * E1SPIRPRP+D57G+N94K+D96L+L97M+ 27% 53% Q249Rπ Lumafast™ (Ps. mendocina) 3% 3% Lipomax™ (Ps. pseuodoalcatigenes L21M) 1 % 0% Fusarium solanipisi 0% 1% Lipolase 0% 0% a) Enzyme Inactivated Commercial European Detergent Lipolytic Enzyme % removal at 1250 2500 12500 LU/1 LU/1 LU/1 SPIRR+D57G+N94K-I-D96L+L97M+Q249R * 12% n.d. 38% SPIRR+N94K+D96L+Q249R* 13% n.d. 45% SPIRR+I90F+D96L+E99K+V187A* n.d. 27% 48% SPIRR+D137G+D167G+D210V+W221L* 13% n.d. 47% SHSRHNA+D57G+N94K+D96L+L97M+Q249R* n.d. 22% 53% GP-rRPRP+D57G+N94K+D96L+L97M+Q249R* n.d. 26% 54% TAIRPRK+D57G+N94K+D96L+L97M+Q249R* n.d. 34% 55% b) Enzyme Inactivated Commercial European Detergent Lipolytic Enzyme % remov al at 0.25 2.50 mg/l mg/l I90F+D96L+E99K+V187AQ 1% 26% ElPSPIRPR+D57G+N94K+D96L+L97M+Q249Rπ 21% 51% I c) Enzyme Inactivated Commercial European Detergent Lipolytic Enzyme delta Reflectance (dR) 1250 5000 12500 LU/1 LU/1 LU/1 A47V+D92G+D96L+A121Tπ 1 n.d. 2 D57G+N94K+D96L+P256Tπ 0 n.d. 2 N94K+D96A+Q249Rπ 0 n.d. 2 SPIRR+Lipolase™* n.d. n.d. 3 D57G+G59V+N94K+D96L+L97M+S116Pπ 1 n.d. 3 D57G+N94K+D96L+L97M+D167G+E210Vπ 1 n.d. 3 QPIRR+D57G+N94K+D96L+L97M+Q249Rci 1 n.d. 3 SPIR+D57G+N94K+D96L+L97M+Q249Rπ n.d. n.d. 4 SHWQQ+D57G+N94K+D96L+L97M+Q249Rπ 1 n.d. 4 I90F +D96L+E99K+ VI 87A +D234Yπ 1 4 n.d. E1AWWPSPIRPRP+D57G+N94K+D96L+L97M+ 2 6 n.d. Q249Rπ SPIRR+A19T+D167G+E210V+W221L* 1 n.d. 6 SPIRR+D57G+N94K+D96L+P256T* 1 n.d. 6 SPIRR+E56K+D57G+D62R+S83T+S85F+ 4 n.d. 11 D96L+D102Y+E210K* SPIRR+N94K+F95L+D96H+N101S+F181L+ 4 n.d. 11 D234Y+Y252L+P256T+G263A+L264Q* SPIRR+D57G+G59V+N94K-I-D96L+L97M+S116 5 n.d. 11 P+S170P+Q249R* SPIRR+A49P+D167G+E210V* 3 n.d. 12 SPIRR+N94K+D96L+Q249R* 4 n.d. 13 SPIRR+D137G+D167G+E210V-I-W221L* 5 n.d. 13 SPIRR-I-D57G+N94K+D96L-I-L97M-I-Q249R* 6 n.d. 13 SPIRR+I90F+D96L+E99K+V187A* 6 n.d. !5 d) Enzyme Inactivated Commercial European Detergent Lipolytic Enzyme delta Reflectance (dR) 0.25 1.00 2.50 mg/l mg/l mg/l D57G+N94K+D96L+L97M+D167G+E210Vπ 1 n.d 3 S3R.D137G+D167G+E210V+W221Lπ 0 2 2 D57G+N94K+D96L+L97M+E210Kπ n.d. n.d. 3 E1SPPRRP+I90F+D96L+E99K-I-D137G-I-D167G+ n.d. 4 n.d. V187A+Q249Rπ E87K+G91A+D167G+E210Vπ n.d. n.d. 4 E87K+G91A+E210KD 1 n.d. 4 I90F+D96L+E99Kπ 0 2 5 APPPRTRPRPRPR+E1S+V2G+S3T+Q4P+D5E+ 0 2 n.d. D57G+N94K+D96L+L97M+Q249Rα N94K+D96L+L97M+N233R+Q249Rπ 0 3 n.d. SPIRKSPIRR+I90F+D96L+E99K+V187Aπ 3 5 D137G+D167G+E210V+W221L+N233Rπ 3 5 SPIRRSPIRR+I90F+D96L+E99K+V187Aπ 3 6 D167G+E210V+N233R+Q249Rπ 3 n.d. ElW+V2P+N94K+D96L+Q249Rπ 3 n.d. D96L+E99K+V187Aπ 3 n.d. ElSPPWWPRW+N94K+D96L+Q249Rπ 2 3 n.d. N94Kπ 2 3 n.d. D96L+D137G+D167G+E210Vπ 2 3 n.d. E1SQRIKQRIK+I90F-I-D96L+E99K-I-V187AD 0 4 n.d. E1SPPRRP+I90F+D96L+E99K+D137G+D167G+ 0 4 n.d. V187A+Q249Rπ I90F+D96L+E99K+V187A+D234Y+Q249Rπ 0 4 n.d I90F.D96L+E99K+V187A+N233RQ 1 4 n.d ElA+S3R+N94K+D96L+Q249Rπ 1 4 n.d. d) Enzyme Inactivated Commercial European Detergent Lipolytic Enzyme delta Refl ectance (dl ) 0.25 1.00 2.50 mg/l mg/l mg/l S3R+I90F+D96L+E99K+V187A+Q249Rπ 1 4 n.d ElA+I90F+D96L+E99K+V187Aπ 1 4 7 I90F+D96L+E99K+V187Aπ 1 4 8 ElSPIRPRP+D57G-l-N94K+D96Lπ 2 4 n.d. ElSPPWWP.N94K+D96L+Q249Rπ 2 4 n.d. SP K+D57G+N94K+D96L+L97M+Q249Rπ 3 4 10 SPIRRP+D57G+N94K+D96L-l-L97M+Q249Rπ 3 n.d. 11 I90F+D96L+E99K+V187A+Q249Rπ 1 5 8 I90F+D96L+E99K+V187A+T231RO 2 5 n.d. ElSPPRWP+N94K+D96L+Q249Rπ 2 5 n.d. E1SPPRWPWR+N94K+D96L+Q249R° 2 5 n.d. N94K+D96L+E99Kπ 2 5 n.d. E1A+I90F+D96L+E99K+Q249R* 1 6 n.d. ElK+D96L+D167G+E210V+N233R+Q249Rα 2 6 n.d. ElSPIRKPRIK+I90F-l-D96L+E99K+V187Aπ 2 6 n.d. SHWRK+ +D57G+N94K+D96L+L97M+Q249Rπ 3 6 n.d. SPIRKAWWP+I90F+D96L+E99K-l-Vl87Aπ 2 7 10 N94K+D96L+E99K+Q249Rπ 2 7 n+d. ElSPPWRPRR+N94K+D96L+Q249Rα 2 7 n.d. E1SPPRWPRR+N94K+D96L+Q249RO 2 7 n.d. D137G+D167G+E210V+W221L+D234Rπ 2 7 n.d. P-4C+N94K+D96L+E239C+Q249Rπ 3 7 n.d. E1SPIRPRPSPIRPRP-I-D57G-I-N94K+D96L-I-L97M 3 7 n.d. +Q249Rπ E1APPPRPRPRPRP+V2G+D5E+D57G+N94K+ 4 7 n.d. D96L+L97M+Q249R* d) Enzyme Inactivated Commercial European Detergent Lipolytic Enzyme delta Refl ectance (dl 9 0.25 1.00 2.50 mg/l mg/l mg/l ElSPPWPRPRP+N94K+D96L+Q249Rπ 2 8 n+d. ElSPKRKPRP+D137G+D167G+E210V+W221Lπ 3 8 n.d. E1SPPRRP+D96L+E99K+D137G+D167G+ 4 8 n.d. V187A+Q249Rπ ElSPPRRP+D57G+N94K+D96L+Q249Rπ 4 9 11 ElSPIRPRP+N94K+D96A-l-Q249Rπ 4 9 n.d. ElSPPRRP+D57G+N94K+D96L+L97M-l-Q249Rπ 5 9 n.d. ElSPPRRP+I90F+D96L+E99K+D137G+V187Aπ 5 9 n.d. E1SPPRRP+Y53C+D57G+N94K+D96L+K127C+ 5 9 n.d. Q249Rπ E1SPPRRP+I90F+D96L+E99K+D137G+V187A+ 4 10 n.d. Q249Rπ E1SPPRRP+N94K+D96L+Q249R Q 5 10 n.d. ElSPPRRP+N94K+D96L+E99Kπ 5 10 n.d. E1SPPRRP+N94K+D96L+E99K+Q249RH 5 10 n.d. ElSPIRPRP+D57G+N94K+D96L+Q249Rπ 6 10 13 ElSPPRRP+I90F+D96L+E99K+V187Aπ 6 10 15 ElSPIRPRP+N94K+D96L+L97M+Q249Rπ 6 10 n.d. E1SPPRPRP+N94K+D96L+Q249R Q 6 10 n.d. APPPRPRLLPIS+D5E+D57G+N94K+D96L+ 6 10 n.d. L97M+Q249R* ElSPIRPRP+D137G+D167G+E210V+W221Lπ 7 10 13 ElSPPPRPRP+N94K+D96L+L97M+Q249Rπ 7 10 n.d. E1SPIRPRP+N94K+D96L+Q249RD 7 11 n.d. ElSPIRPRP+D57G+N94K+D96L+L97M+Q249Rπ 7 13 16 EXAMPLE 8 The first wash activity of one first wash Upolytic enzyme was tested using the "Assay for test of First Wash effect" described in the Materials and Methods section above with an array of commercial detergents. The enzymes already present in the detergents were inactivated by heat (4 minutes at 85° C in microoven) prior to wash. The Lipolase variant E1SPIRPRP+D57G+N94K+D96L-I-L97M-I-Q249R produced in Aspergillus oryzae as described in example 2 was used. The following different geographic condtions were used: European: Time: 20 min. Temperature: 30°C Water hardness: 3.2 mM Ca 7Mg 2 - (5:1) ~18°dH US: Time: 10 min. Temperature: 30°C Water hardness: 1.07 mM Ca 2 7Mg 2 ' (5:1) ~ 6°dH E1SPIRPRP+D57G + N94K + D96L + L97M + Q249Rπ in US detergents Detergent dR at dR at 0.25mg/l 1.00 mg/l Wisk HDL (2 g/1) 3 5 Wisk w. bleach (1 g/1) 3 7 Surf w. bleach (1 g/1) 1 4 Tide HDL (2 g/1) 1 4 Tide w. bleach (1 g/1) 2 5 E1SPIRPRP+D57G + N94K + D96L + L97M + Q249Rπ in European detergents Detergent dR t dR at 0.25mg/l 1.00 mg/l Ariel Futur (5 g/1) 6 12 TJBA 06731122 Ariel Futur color (5 g/1) 7 11 UBA 06730101 TandU Ultra Plus (5 g/1) 4 12 UBA 02500191 Tandil Ultra Plus Color (5g/l) 5 13 UBA 05761612 Sunil Aktiv (5.5 g/1) 4 15 UBA 05580168 Suml Aktiv Citrus (5.5 g/1) 3 13 UBA 05580168 SunU Aktiv Color (5.5 g/1) 4 13 UBA 05580169 Persil Megapearls (5 g/1) 2 12 UBA 04163661 Persil Megapearls Color (5 g/1) 3 16 UBA 04163662 EXAMPLE 9 PRODUCTION OF A FIRST WASH LIPASE IN F. GRAMINARUM Strains and Media The starting strain is Fusarium graminearum A3/5 (ATCC 20334). COVE plates are comprised of 343.3 g of sucrose, 20 ml of COVE salts solu¬ tion, 10 ml of 1 M acetamide, 10 ml of 3 M CsCl, and 25 g of Noble agar per Uter. The COVE salts (50X) solution is comprised of 26 g of KCI, 26 g of MgSO 4 -7H 2 O, 76 g of KH 2 PO 4 , and 50 ml of COVE trace metals solution. COVE trace metals solution is com- prised of 0.04 g of NaB 4 O 7 -10H 2 O, 0.040 g of CuSO 4 -5H 2 O, 0.70 g of FeSO 4 -H 2 O, 0.80 g of Na 2 MoO 2 -2H 2 O, and 10 g of ZnSO 4 per Uter. M400Da medium is comprised of 50 g of maltodextrin, 2.0 g of MgSO 4 -7H 2 O, 2.0 g of KH 2 PO 4 , 4.0 g of citric acid, 8.0 g of yeast extract, 2.0 g of urea, and 0.5 ml of trace metals solution per Uter. The medium is adjusted to pH 6.0 with 5 N NaOH. The trace metals solution is comprised of 14.3 g of ZnSO 4 -7H 2 O, 2.5 g of CuSO 4 -5H 2 O, 0.5 g of NiCl 2 -6H 2 O, 13.8 g of FeSO 4 -7H 2 O, 8.5 g of MnSO 4 -H 2 O, and 3.0 g of citric acid per Uter. Construction of the H. lanuginosa lipolytic enzyme variant HL A (E1SPIRPRP+D57G+N94K+D96L+L97M+Q249R) expression plasmid for Fusarium graminearum The construction an expression plasmid for Fusarium graminearum is outlined in Fig. 7. SpecificaUy, a Fusarium expression cassette is made using the technique of overlapping PCR (Higuchi. R., In Innis, M.A.. Gelfond, D.H., Snisky. J.J., and White. TJ., editors. PCR Protocols: A Guide to Methods and Applications, pages 177-183,. Academic Press, Inc., New York) to fuse the 1.24 kb Fusarium oxysporum trypsin pro¬ moter to the 1.1 kb Fusarium oxysporum trypsin terminator (Royer et al., 1995 Bio/Technology 13: 1479-1483). A polyUnker region containing Swal, Kpnl and Pad restriction sites is inserted between the promoter and terminator as part of the overlapping PCR reaction. At the 5'end of the promoter an Xhol site is added and the native EcoRI site is preserved. At the 3' end of the terminator, EcoRI, Hindlll and Nsil sites are in¬ corporated by the PCR reaction. A PCR fragment containing -1208 to -1 of the Fusarium oxysporum trypsin promoter plus a 25 bp polylinker is generated from plasmid pJRoy20 (Royer et al , 1995, supra) using the following primers: Xhol .EcoRI 5 'end of promoter Forward primer 1: 5'-gagctcgagGAATTCTTACAAACCTTCAAC-3' Pacl Kpnl Swal 3'end of promoter Reverse primer 2 : 5 ' - ttaattaaggtacctgaatttaaatGGTGAAGAGATAGATATCCAAG-3' Upper case letters are the native sequence of the Fusarium oxysporum trypsin promoter. The PCR conditions used are 95°C for 3 minutes followed by 25 cycles each at 95°C for 30 seconds, 50°C for 1 minute, and 72°C for 1 minute. The final extension cy¬ cle is at 72°C for 5 minutes. Pwo DNA polymerase (Boehringer Mannheim, Indianapo- 5 Us, IN.) is used with the manufacturer's supplied buffer. A PCR fragment containing -5 to -1 of the Fusarium oxysporum trypsin pro¬ moter, the 25 bp polylinker and 1060 bp of the 3' untranslated region of the Fusarium oxysporum trypsin gene (terminator region) is generated from plasmid pJRoy20 using the foUowing primers: 10 promoter Swαl Kpnl Pacl 5' end of terminator Forward primer 3: 5'-TCACCatttaaattcaggtaccttaattaaATTCCTTGTTGGAAGCGTCGA- 3' 15 N-sil Hindm EcoRI 3' end of terminator Reverse primer 4: 5'-tggtatgcataagcttgaattcAGGTAAACAAGATATAATTT-3' Upper case letters are the native sequence of the Fusarium oxysporum trypsin promoter and terminator. The PCR conditions used are as described above. 20 The final 2.3 kb overlapping PCR fragment which contains -1208 to -1 of the Fusarium oxysporum trypsin promoter, the 25 bp polylinker and 1060 bp of the Fusarium oxysporum trypsin terminator is made using 0.2 μl of the first PCR (promoter) reaction and 3 μl of the second (terminator) reaction as template and primers number 1 and 4. The PCR conditions used are 95°C for 3 minutes followed by 30 cycles each at 95°C for 25 30 seconds, 62°C for 1 minute, and 72°C for 3 minutes. The final extension cycle is 5 minutes at 72°C. Pwo DΝA polymerase is also used for this reaction. The resulting 2.3 kb band is digested with Xhol and Nsil and cloned into plasmid pBaΝeό that is digested partially with Nsil and to completion with Sail. In effect, the Aspergillus promoter and terminator sequences of pBaΝeό are replaced with the Fusarium 30 oxysporum trypsin promoter and terminator sequences. The resulting construct (pDM174.3) is digested with Swal and Pacl. DNA primers HLIP-A and HLIP-B shown below are used in a PCR reaction to amplify the HLA Upase gene from plasmid pJVi220: HLIP-A (Primer 5): 5'-cccatttaaatATGAGGAGCTCCCTTGTGCTG-3' HLIP-B (Primer 6): 5'-cccttaattaaCTAAAGACATGTCCCAATTAA-3' Uppercase letters represent sequences in the lipase gene The PCR is performed in a 50 μl reaction containing ca. 50 ng of pHLA, 0.05 mM each of dATP, dTTP, dGTP, dCTP, 100 pmol each of HLIP-A and HLIP-B. IX -Pvvol Buffer (Boehringer Mannheim, Indianapolis, IN), and 2.5 units Pwol (Boehringer Mannheim, Indianapolis, IN). The PCR conditions are 95°C for 3 minutes, 30 cycles each at 95°C for 1 minute, 60°C for 1 minute; and 72°C for 1.5 minutes, and then 72°C for 5 minutes. The PCR reaction mixture is run on a agarose gel and the ca. 0.9 kb HLA DNA band is excised. The DNA is purified by solubilization of the agarose with 3 vol¬ umes Qia-ex solubUization buffer (Qiagen, Los Angeles, CA) followed by a Qiaquick PCR spin column according to the manufacturer's directions (Qiagen, Los Angeles, CA). The DNA is recovered in 50 μl of 1 mM EDTA-10 mM Tris pH 8. A 20 μl aUquot of the DNA is cut in a final volume of 25 μl containing IX restriction enzyme buffers and restriction enzymes Pad and Swal as suggested by the manufacturers. The reaction mixture is then heated at 80°C for 10 minutes. One μl of the Pa / Swal cut HLA lipase gene is ligated into Pad/ Swal cut plasmid pBANeό. The ligation mixture is used to transform E. coli strain DH5α. The plasmid containing pBANeό and the HLA sequences is designated pJeRS33. The 0.9 kb Swάl/Pacl HLA fragment from pJeRS33 is cloned into the Swάl/Pacl digested pDM174.3 vector to create plasmid pDM177. Transformation of Fusarium graminearum F. graminearum strain A3/5 (ATCC 20334) is grown on 10 x 15 mm petri plates of Vogels medium (Vogel, 1964, Am. Nature 98: 435-446) plus 1.5% glucose and agar for 3 weeks at 25°C. Conidia (approximately IO 8 per plate) are dislodged in 10 ml of sterile water using a transfer loop and purified by filtration through 4 layers of cheese- cloth and finally through one layer of Miracloth. Conidial suspensions are concentrated by centrifugation. Fifty ml of YPG medium comprised of 1 % yeast extract, 2% bac- topeptone, and 2% glucose are inoculated with approximately 100 conidia, and incubated for 14 hours at 24°C, 150 rpm. Resulting hyphae are trapped on a sterile 0.4 mm filter and washed successively with sterile distilled water and 1.0 M MgSO 4 . The hyphae are resuspended in 10 ml of NOVOZYM 234™ (Novo Nordisk A/S, Bagsvaerd, Denmark) solution (2-10 mg/ml in 1.0 M MgSO 4 ) and digested for 15-30 minutes at 34°C with 5 agitation at 80 rpm. Undigested hyphal material is removed from the resulting protoplast suspension by successive filtration through 4 layers of cheesecloth and through Miracloth. Twenty ml of 1 M sorbitol are passed through the cheesecloth and Miracloth and com¬ bined with the protoplast solution. After mixing, protoplasts (approximately 5 x 10 8 ) are pelleted by centrifugation and washed successively by resuspension and centrifugation in 10 20 ml of 1 M sorbitol and in 20 ml of STC (0.8 M sorbitol, 0.05 M Tris pH 8.0, 0.05 M CaCl 2 ). The washed protoplasts are resuspended in 4 parts STC and 1 part SPTC (0.8 M sorbitol, 40% PEG 4000, 0.05 M Tris pH 8.0, 0.05 M CaCl 2 ) at a concentration of 1-2 x lOVml. One hundred μl of protoplast suspension are added to 3 μg pDM177 DNA and 5 μl heparin (5 mg/ml in STC) in polypropylene tubes (17 x 100 mm) and incubated on ice 15 for 30 minutes. One ml of SPTC is mixed gently into the protoplast suspension and in¬ cubation is continued at room temperature for 20 minutes. Fifteen ml of molten solution (cooled to 40°C) consisting of COVE salts, 0.8 M sucrose and 1 % low melting agarose (Sigma Chemical Company, St. Louis, MO) are mixed with the protoplasts and then plated onto a 150 mm petri plate containing COVE agar. Incubation is continued at room 20 temperature for 10 to 14 days. Expression of lipase activity Five pDM177 transformants of Fusarium graminearum A3/5 are cultured on M400Da medium in shake flasks for 7 days at 30°C. As a control culture, an A3/5 trans- 5 formant of plasmid pDM155 (Royer et al., 1995 Bio /Technology 13: 1479-1483), which contains the wild-type Humicola lanuginosa lipase inserted between the Fusarium ox¬ ysporum trypsin promoter and terminator, is grown at the same time under the same con¬ ditions. 30 First wash activity of HL A The above produced Upase as weU as the analogous Upase (carrying the same mutations, but produced in A. oryzae) was tested in the "Assay for test of First Wash effect" described herein using enzyme inactivated Ariel Futur. The foUowing results were obtained: ElSP-TRPRP+D57G-l-N94K-l-D96L-(-L97M+Q249R in dR at 1250 LU/1 12500 LU/1 A.oryzae 8 15 F.graminearum 9 14 EXAMPLE 10 - Construction of Absidia reflexa mutants Material and Methods Strains and plasmids are Usted in the Materials and Methods section preceeding the ex¬ amples. Primers: primer TiK57, primer TiK58 , primer TiK59, Primer TiK60, Primer TiK 61 , primer TiK62, primer TiK64, primer Tik66, primer TiK72, primer TiK74, primer Tik75, primers TiK76. Kits, solutions, media and the like: Taq-DNA polymerase (Promega) LB medium supplemented with 100 /-.g/ml ampicUlin DNA Maxi-Prep kit (QIAGEN). Polyethyleneglycol/LiOAc yeast transformation kit (Yeastmaker, Clontech). Yeast nitrogen base w/o amino acids (Difco 0919-15-3) Casamino acids (Difco 0230-01-1) Bacto-agar (Difco) YPG-medium (20 g casein-peptone and 10 g yeast extract (Difco)) LB medium supplemented with 100 μg/ml ampicilUn Equipment Applied Biosystems 373 DNA Sequencer CeUulose acetate filters (Schleicher-Schϋll) SYC-plates 13.6 g NaOH and 22.6 g succinic acid are completely dissolved in 500 ml H 2 O, 15 g yeast nitrogen base w/o amino acids (Difco 0919-15-3), 10 g casamino acids (Difco 0230-01-1). 20 ml of a 2 % threonine solution, and 20 ml of a 1 % tryptophan solution are added. The solution is filled up to 1 litre H 2 O. sterile-filtered, and stored at 4°C. For liquid media preparation 1 Utre of SYC is diluted by adding 200 ml of a 20% glucose solution and 800 ml of H 2 O. For preparation of agar plates 1 Utre of SYC is dUuted at 60°C with 800 ml of bacto-agar (37.5 g bacto-agar (Difco) dissolved in 1 Utre H 2 O) and 200 ml of a 20% glucose solution. Brilliant Green assay For preparation of BG-agar plates 10 g of agarose are dissolved by heating in 500 ml of a 100 mM Tris-CI buffer pH 9.0. This agarose solution is mixed at 60 °C with 24 ml of an olive oil emulsion (8 ml olive oil (Sigma) and 16 ml of a 2% polyvinylalco- hoi (Sigma) solution in H 2 O are mixed and thoroughly homogenized on ice with an Ultra- Turrax T25), 10 ml of a BriUiant Green stock (4 mg/ml H 2 O), 465 ml of 100 mM Tris¬ Cl buffer pH 9.0 containing 6.45 g of the detergent, and 1.4 ml of a 250 mM CaCl 2 so¬ lution. Transformed yeast colonies are grown for 3 days at 30 °C on sterile cellulose acetate filters (Schleicher-SchuU) on SYC-agar plates. The filters are then transfeπed to the BG-agar plates and incubated for 2-8 hours at 37° C. Colonies which generate a green spot in the agar are judged positive. Restriction enzyme analysis and sequencing of mutant libraries The recovered plasmid DNA was subjected to Nco/-digestion. Conect clones yielded two fragments of 2.9 and 3.1 kb. The plasmid DΝA was subsequently either dye- terminator PCR cycle sequenced or sequenced on an Applied Biosystems 373 DΝA Se¬ quencer. In the case of clones isolated from libraries 3 and 4 (Table 1) the oligonucleo¬ tides TiK61 and TiK66 were used for double-strand sequencing whereas for libraries 7 and 8 TiK62 and TiK72 were appUed. Example 10A Construction of Absidia reflexa ATTC 44896 lipase expression vectors Two vectors were constructed for expression of the wild-type Absidia reflexa ATTC 44896 lipase (with/without SPIRR peptide extension) in Saccharomyces cere- visiae. The cDNA clone encoding the mature Absidia reflexa ATTC 44896 Upase (SEQ ID No. 13 and Figure 1) was PCR amplified (T, = 50°C, 25 cycles, 5 units Taq-DNA polymerase) using either the primer pair TiK57/59 (providing a N-terminal SPIRR exten¬ sion) or the primer pair TiK58/59 (without SPIRR extension). The PCR fragments were Ugated via Nhel/Xbal-sites into the yeast expression vector pJSO37 (with a Nhel site in¬ troduced downstream of the signal peptide) which harbours region encoding an active H. lanuginosa Upase in the original BamΗI/Xbal cloning site. The gene encoding the active H. lanuginosa Upase was slightly modified by the in¬ troduction of a Nhel-site downstream of the BamΗI site (see Figure 2). Since Nhel- and Xbal-sites are compatible, the vector was treated with alkaUne phosphatase prior to Uga- tion. Finally, two vector constructs were obtained, namely pTiK04 which includes the SPIRR extension just upstream of the start of the mature lipase gene, and pTiK05 which did not contain a SPIRR extension encoding part. In both cases the original signal se¬ quence from the Humicola lanuginosa lipase was kept constant between the BamHI and Nhel sites. MFαl signal sequence constructs The H. lanuginosa lipase signal sequence was replaced by the mating factor αl signal. Genomic DNA of the yeast strain YPΗ499 (Stratagene) was prepared cording to standard protocols (Ausubel et al., (1995). Cuπent Protocols in Molecular Biology, John Wiley and Sons). One μg of genomic DNA was subjected to PCR (T a = 50°C, 25 cycles, 5 units Taq-DNA polymerase) with the primer pair TiK74/75. The amplified MFαl signal sequence fragment (Fig. 3) was inserted into pTiK04 and pTiK05 via the BamHI and Nhel sites to yield pTiKOό and pTiK07, respectively. Example 10B Construction of Absidia reflexa ATTC 44896 lipase variants by mutagenesis with doped oligonucleotides Four libraries were constructed with doped oligonucleotides. In Ubraries A and B (see Table 1 below)) random mutations were introduced in the putative lid region of Absidia reflexa ATTC 44896 lipase. In library A the SPIRR se¬ quence was kept constant, in Ubrary B it was omitted. Libraries C and D were constructed with mutations in two putative Upid contact zones in the N-terminus of Absidia reflexa ATTC 44896. In library C the SPIRR se¬ quence was again kept constant. The mutagenesis of the putative lid region of Absidia reflexa ATTC 44896 lipase gene (amino acid positions 82-99) was performed by standard PCR (Sambrook et al., (1989), Molecular cloning - A laboratory manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor. NY) with 5 units Taq-DNA polymerase with the primer pair TiK60/TiK64 at T a = 50°C and 25 cycles using pTiK05 as template. In a second stan- dard PCR, using primer TiK62 and the agarose gel-purified DNA fragment generated in the first PCR, the whole Absidia reflexa ATTC 44896 lipase gene was restored using the same amplification conditions as for the first PCR. In contrast to the first PCR, pTiK04 or pTiK05 were chosen as templates so that libraries (with/without SPIRR N-terminal extension) were obtained. For mutagenesis of the amino acid positions 30-45 the primers TiK76 and TiK64 were used for the first PCR. The second PCR was in principle identical to the above de¬ scribed one. The obtained PCR fragments were Ugated into pJSO37 via the BamHI/Xbal-sites. The transformation of E. coli and subsequently of competent YNG318 yeast cells was performed as described above. Table 1 Summary of the constructed and screened Absidia reflexa ATTC 44896 mutant Ubraries. No Comments Library Screened Positives in Positives in Restric¬ and muta¬ size in S. cerevisi¬ the BG-assay the BG-assay tion en¬ genized re¬ E.coli ae colonies at X g/1 de¬ at X g/1. zyme gions tergent 2.round analysis A Lid (pos. 82- 8 x l0 6 180000 21 at 3g/l 9 at 3 g/1 6 Example IOC Screening of the Absidia reflexa ATTC 44896 lipase mutant libraries and recovery of plasmids from positive colonies Transformed yeast ceUs obtained as described in Example 10B were spread on a cellulose acetate filter on a 14 cm SYC-agar plate. After selective growth of transformants and transfer of the filter to BG-agar plates, positive colonies were picked, resuspended in H 2 O, and re-spread on cellulose acetate filters on SYC-agar plates to allow a second con- firmatory round of the BG-assay (see Matarials and Methods section). Positive clones from the second round were transfeπed to 20 ml SYC-medium and shaken for 2 days at 30°C. Plasmid DNA was prepared from 1.5 ml of this saturated yeast culture according to the standard phenol/glassbead method (Ausubel et al., (1995), Cuπent Protocols in Molecular Biology, John WUey and Sons). An aliquot of the plasmid preparation was electroporated into E.coli DH10B ceUs which were then plated on LB-agar plates supplemented with 100 μg/ml ampicUlin. The DNA of the colonies was isolated and applied to restriction enzyme analysis and sequencing as described in the Material and Methods section above. Sequencing Sequencing of 15 randomly picked clones in Ubrary A and B showed that the chosen doping of 10% finally resulted in: 20% being wild type, 13% with 1 amino acid exchange, 20% with 2 exchanges, 13% with 3, 20% with 4, 0% with 5, and 13% with 6 amino acid exchanges per molecule. From library A six clones out of 1.8 x 10 s screened colonies at 3 g/1 detergent were iso¬ lated in the BG-assay (1 per each 3 x IO 4 screened), and from Ubrary B two positive clones were isolated out of 2.0 x 10 s screened colonies (1 per each 1.0 x IO 5 screened, Table 1). The noticed sequence differences of these 8 clones are depicted in Table 2 be¬ low. As mentioned above Ubraries C and D were constructed with mutations in two putative lipid contact zones in the N-terminus of Absidia reflexa ATTC 44896 lipase. Sequencing of 13 randomly picked clones showed that the chosen doping level of 10% resulted in: 8% being wild type, 15% with 1 amino acid exchange, 46% with 2 exchanges, 15% with 3, and 15% with 4 exchanges per molecule. Library C yielded 3 clones out of 1.6 x 10 s screened colonies at 3 g/1 of deter¬ gent in the BG assay (1 positive per each 53333 screened), and from library D one posi¬ tive clone could be isolated out of 1.7 x 10 s (Table 1). The sequences of these 4 clones are also depicted in Table 2. Table 2 Sequences of the improved Absidia reflexa ATTC 44896 variants. Target sequence: 82 99 Library Clone number T S I R N A I A P I V F V P V N Y A 303 w T 309 H T 312 S s 315 V T W L -N L H133R 318 C W K L S I ..V102F 321 S E B 401 S S A 402 ..Y136H.K137H 30 45 R T V I P G G R W S C P H C G C 701 W N 702 W N 703 ...Q4R D 801 .V95E Example IOD Expression of Absidia reflexa ATTC 44896 variants and lipase unit measurement Four of the improved mutants identified from library A were subjected to meas- urement of LU secreted to the culture medium as well as LU measurement of the crude cytosol/membrane fraction. 10 ml of YPG-medium were dissolved in 900 ml H 2 O, autoclaved and 100 ml of a 20% glucose solution added) were inoculated with 1 ml of a saturated yeast culture in SYC-medium. The culture was shaken at 30°C for 2 days. The cells were harvested by centrifugation (5 minutes x 4000 g) and the supernatant stored on ice for LU measure¬ ment. The ceU peUet was treated with Novozym™234 for spheroplast preparation and lysed using the glass bead procedure (Ausubel et al., (1995). Cuπent Protocols in Mo¬ lecular Biology, John Wiley and Sons). The obtained crude cytosol fraction which also included the membrane fraction was immediately applied to LU measurement (see Mate- rials and Methods section) in order to minimize proteolytic degradation. The result is shown in Table 3. The four clones showed weak but significant LU in the cytosol/membrane fraction. Moreover, from two out of these four clones weak LU signals could be recorded. All data differing from 0.0 LU/ml are significant enzyme ac¬ tivities as confirmed by repeated measurements. Table 3 Summary of the obtained lipase unit secreted to the medium or measured from the cytosol fraction of ATTC 44896 variants from Ubrary A. The standard deviation is 10%. Sample Lipase units in the Lipase units in the supernatant (LU/ml) cytosol (LU/ml) YNG318 (negative control) 0.0 0.0 303 0.0 1.0 309 0.0 0.5 312 1.7 0.6 321 0.6 1.0 EXAMPLE 12 Substrate affinity of Lipolytic enzymes A procedure has been developed for a simple comparison of the ability of Upolytic enzymes to accumulate on/in a substrate phase (oUve oil, incl. FFA) at alkaline pH (pH 9.0) and presence of the non-ionic surfactant Dobanol 25-7 (100 ppm) (i.e. a measure for substrate affinity). Procedure: 1. Two identical buffer solutions (5 ml) are prepared in 20 ml sealable vials, ("Sample" (s) and "Reference" (r)). 2. Enzyme is added into "Sample" and "Reference" and the lipase concentration is determined (X LU/ml). 3. Olive oil is added onto the "Sample" and both lipase solutions are shaken vigorously. Incubation at 4°C over night. 4. Remaining lipase concentration in the aqueous phases is determined after incubation, (Yi LU/ml; i=r,s). Summary of incubation conditions; Buffer: 100 mM Glycine (5 ml). pH: 9.0. Substrate: Olive oil (5 ml). Dobanol 25-7: 100 ppm. T: 4°C. Lipase: 5-10 LU/ml. Incubation: Over night (24-26 hours) Evaluation of data: The result after an experiment is calculated by comparing the activity-loss upon incuba¬ tion in the aqueous phase in contact with olive oU to the activity-loss in the aqueous phase in absence of oUve oil: α = Ys/Yr (see above) Results: Table 11: Lipase α (%) Lipolase™ 95 % D57G+N94K+D96L+L97M+Q249R 65 % SPIRPR+E1P+D57G+N94K+D96L+L97M+Q249R 45 % SALRPRK+D57G+N94K+D96L+L97M+Q249R 25 % SPIRPR+E1P+D137G+D167G+E210V+W221L 50% Comparing results presented above to wash data disclosed in examples 5-8, clearly indi¬ cate that Lipolase variants with increased first wash performance generally have increased substrate affinity as compared to Lipolase. Example 13 Localized random mutagenesis of the Pseudomonas sp. lipase fLiposam . A suitable doping scheme to use for introducing mutations contemplated to lead to a first wash activity of the above lipase may comprise localized random mutagenesis in the whole or parts of one or more of the foUowing regions. 93 % wt/7% random means that the respective codons are synthesized with 93% wt nucleotides and 7% of the other 3 nu¬ cleotides in the oligonucleotide used for constructing the random mutagenized Ubrary. Similarly for 90% wt/10% random. In the amino acid region 17-37: Amino acid position 17-18+20-24+26-29+32-37: 93% wt/7% random M19: doped to give preferentiaUy L,I,F In the amino acid region 109-161: Amino acid position 109-118: 93% wt/7% random Amino acid position 120 + 123-137 + 139-161: 90% wt/10% random In the amino acid region 208-239: Amino acid position 208-212+214-215+217-231+233-239: 90% wt 10% random In the amino acid region 253-271: Amino acid position 253+255+259-268+270-271: 90% wt 10% random V258: 90% wt/10% random but doped not to be a positive charged amino acid. The localized random mutagenesis may be performed as described in the Materials and Methods section and in Example 1 herein, and the resulting mutants screened for a re- duced dependence to calcium and/or an increased tolerance towards a detergent or deter¬ gent component and afterwards first wash activity. Subsequently, and if necessary, local¬ ized random mutagenesis of the resulting mutants may be repeated and/or the genes may be subjected to gene shuffling as disclosed herein. E SHEET REFERENCES CITED IN THE SPECIFICATION Muhlrad et al., 1992, Yeast, Vol. 8, 79-82 Shimada. Y, et al. (1989), cDNA Molecular Cloning of Geotrichum candidum Lipase, J. Biochem., 106, 383-388, HunkapUler et al., 1984, Nature 310:105-111 R. Higuchi, B. Krummel, and R.K. Saiki (1988), A general method of in vitro preparation and specific mutagenesis of DNA fragments: study of protein and DNA interactions, Nucl. Acids Res. 16:7351-7367, Yamaguchi. S, et al. (1991), Cloning and structure of the mono- and diglycerol tipase- encoding gene from PeniciUium camembertii U-150, Gene 103, 61-67, Hass. M.J. et al. (1991), Clomng, expression and characterization of a cDNA encoding a Upase from Rhizopus delemar, Gene 109. 107-113, Kugimiya. W. et al. (1992), Cloning and Sequences Analysis of DNA encoding Rhizopus niveus Lipase, Biosci. Biotech. Biochem. 56, 716-719. Dartois. V. et al. (1993), Cloning, nucleotide sequence and expression in Escherichia coti of a Upase gene from Bacillus subtilis 168, Biochemica et Biophysica acta 1131, 253-260, Sambrook et al. Molecular Cloning: A Laboratory Manual, 2nd Ed., Cold Spring Harbor, 1989, R.K. Saiki et al., Science 239, pp. 487-491, 1988, Beaucage and Caruthers, Tetrahedron Letters 22, pp. 1859-1869, 1981, Matthes et al. , The EMBO J, 3, pp. 801-805, 1984, J.O. Deshler, (1992) A simple method for randomly mutating cloned DNA fragments by using chemical mutagens and the polymerase chain reaction, GATA 9(4): 103-106 Leung et al., Technique, Vol. 1, No. 1, pp. 11-15, 1989 Fowler et al., Molec. gen. Genet., 133, pp. 179-191, 1974, Brady et al., "A Serine Protease Triad Forms the Catalytic Centre of a Triacylglycerol Lipase", Nature 343, 1990, pp. 767-770, 1990, TUbeyrgh. H, van Egloff, M.-P., Martinez.C, Rugani.N., Verger.R. and CambUlau (1993) Nature 362, p. 814-820, Interfacial activation of the Upase-proUpase complex by mixed miceUes revealed by X-ray crystaUography. Hudson et al., Practical Immunology, Third edition, BlackweU Scientific PubUcations, 1989 Alber. T. and Kawasaki, G., J.Mol.Appl. Genet i, 419-434 (1982) Joyce, G. F.: Directed Molecular Evolution. Scientific American December 1992, pp. 48-55. Siderovski, D. P., and Mak T. W.: RAMHA: A pc-based monte-carlo simulation of ran¬ dom saturation mutagenesis. Comput. Biol. Med. 23(6), pp. 463-474, 1993. PalzkUl T., Le Q.-Q., Venkatachalam K. V., LaRocco M., and Ocera H.:Evolution of antibiotic resistance: several different amino acid substitutions in an active site loop alter substrate profile of b-lactamase. Molecular Microbiology 12(2), pp. 217-229, 1994. Oltis, D.L. et al (1992) Prot. Eng. 5, p. 197. Svendsen, A. et al. (1995) Biochim. Biophys. Acta. 1259, p. 9. Grochulski, P. et al. (1993) J. Biol. Chem. 268, p. 12843. SEQUENCE LISTING (1) GENERAL INFORMATION: (i) APPLICANT: (A) NAME: Novo Nordisk A/S (B) STREET: Novo Alle (C) CITY: Bagsvaerd (E) COUNTRY: Denmark (F) POSTAL CODE (ZIP) : DK-2880 (G) TELEPHONE: +45 4444 8888 (H) TELEFAX: +45 4449 3256 (ii)TITLE OF INVENTION: Novel Lipolytic enzyme (iii)NUMBER OF SEQUENCES: 100 (iv) (iv) COMBUTER READABLE FORM: (A) MEDIUM TYPE: Floppy disk (B) COMPUTER: IBM PC compatible (C) OPERATING SYSTEM: PC-DOS/MS-DOS (D) SOFTWARE: Patentin Release #1.0, Version #1.30B (EPO) (2) INFORMATION FOR SEQ ID NO: 1: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (A) DESCRIPTION: /desc = "Primer 3" (xi) SEQUENCE DESCRIPTION: SEQ ID No. 1: AGAAATCGGG TATCCTTTCA G 21 (2) INFORMATION FOR SEQ ID NO: 2: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 30 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (A) DESCRIPTION: /desc = "Primer 7258" (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 2: gaatgacttg gttgacgcgt caccagtcac 30 (2) INFORMATION FOR SEQ ID NO: 3: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 26 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (A) DESCRIPTION: /desc = "Primer 7770" (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 3: tctagcccag aatactggat caaatc 26 (2) INFORMATION FOR SEQ ID NO:4: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 53 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (A) DESCRIPTION: /desc = "randon 1" (B) Bottle 5: 93% A; 2.33% C; 2.33% G and 2.33% T Bottle 6: 93% C; 2.33% A; 2.33% G and 2.33% T Bottle 7: 93% G; 2.33% A; 2.33% C and 2.33% T Bottle 8: 93% T; 2.33% A; 2.33% C and 2.33% G (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 4: 5 ' 5 C G T 5 C 3 T 7 . A A 8 G T 8 T T A C T T 5 C C 7 T T 5 C T 8 T T 8 A 6 C/G T 5 6 G 5 6 G 7 G A 8 AA A 6 T C 7 (2) INFORMATION FOR SEQ ID NO: 5: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 93 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (A) DESCRIPTION: /desc = "Oligo 2" Flask 5: 80% A; 6,66% C; 6,66% G og 6,66 % T. Flask 6: 80% C; 6,66% A; 6,66% G og 6,66 % T. Flask 7: 80% G; 6,66% A; 6,66% C og 6,66 % T. Flask 8: 80% T; 6,66% A; 6,66% C og 6,66 % G. (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 5: GTCTCTGCGT GGACGGCCTT GGCGGCGCCA CCTCCA67 (T/A) 66(T/A) 575 66(T/A) 67 (T/A) 66 (T/A) 575 66 (T/A) (6/7) (7/8) (C/G) 57 (C/G) C57 (5/7) 5 (C/G) CTGT TTAACCAGTT CAATCTC 93 (2) INFORMATION FOR SEQ ID NO: 6: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 64 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid HEET (A)DESCRIPTION: /desc = "Oligo lid2" Flask 5: 93% A; 2,33% C; 2,33% G og 2,33 % T. Flask 6: 93% C; 2,33% A; 2,33% G og 2,33 % T. Flask 7: 93% G; 2,33% A; 2,33% C og 2,33 % T. Flask 8: 93% T; 2,33% A; 2,33% C og 2,33 % G. (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 6: CATTTAT886 888655 (C/G) (A/C/G/T) (A/C/G/T) 7 55 (C/G) 88 (A/C) 5758 8 (C/G) 76 (7/8) 58665 788688(8/7)587 75ACGAG(A/T) GC CACG 64 2) INFORMATION FOR SEQ ID NO: 7: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 105 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (A) DESCRIPTION: /desc = "Primer 4" (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 7: GCTCCTCATG GTGGATCCCC AGTTGTGTAT ATAGAGGATT GAGGAAGGAA GAGAAGTGTG GATAGAGGTA AATTGAGTTG GAAACTCCAA GCATGGCATC CTTGC 105 (2) INFORMATION FOR SEQ ID NO: 8: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 54 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (A) DESCRIPTION: /desc = "Primer 8479" (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 8: GCGTGGACGG CCTTGGCTAG CCCTATTCGT CCTCGACCGG TCTCGCAGGA TCTG 54 (2) INFORMATION FOR SEQ ID NO: 9: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 30 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (A) DESCRIPTION: /desc = "Primer 7887" (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 9: GAATGACTTG GTTGAGTACT CACCAGTCAC 30 (2) INFORMATION FOR SEQ ID NO: 10: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 46 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (A) DESCRIPTION: /desc = "Primer 8932" (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 10: GAACTGGATA GGAAATTTGA AGTTCCTGTT GAAAGAAATA AATGAC 46 (2) INFORMATION FOR SEQ ID NO: 11: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 57 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (A) DESCRIPTION: /desc = "Oligo 1" Flask 5: 80% A; 6.66% C; 6.66% G og 6.66 % T. Flask 6: 80% C; 6.66% A; 6.66% G og 6.66 % T. Flask 7: 80% G; 6.66% A; 6.66% C og 6.66 % T. Flask 8: 80% T; 6.66% A; 6.66% C og 6.66 % G. (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 11: GCGTGGACGG CCTTGGCC86 (T/A) 66 (A/T) 58 (T/A) 67 (T/A) 66 (T/A) 575 66 (T/A) GAGGTC TCG CAGGATC TG 57 (2) INFORMATION FOR SEQ ID NO: 12: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (A) DESCRIPTION: /desc = "Primer 2056" (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 12: GCACGTAATG TTTGTACC 18 (2) INFORMATION FOR SEQ ID NO: 13: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (A) DESCRIPTION: /desc = "Primer 4699" (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 13: CGGTACCCGG GGATCCAC 18 (2) INFORMATION FOR SEQ ID NO: 14: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 30 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (A) DESCRIPTION: /desc = "Forward Primer 1" (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 14: GAGCTCGAGG AATTCTTACA AACCTTCAAC 30 (2) INFORMATION FOR SEQ ID NO: 15: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 47 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (A) DESCRIPTION: /desc = "Reverse Primer 2" (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 15: TTAATTAAGG TACCTGAATT TAAATGGTGA AGAGATAGAT ATCCAAG 47 (2) INFORMATION FOR SEQ ID NO: 16: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 51 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (A) DESCRIPTION: /desc = "Forward Primer 3" (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 16: TCACCATTTA AATTCAGGTA CCTTAATTAA ATTCCTTGTT GGAAGCGTCG A 51 (2) INFORMATION FOR SEQ ID NO: 17: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 42 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (A) DESCRIPTION: /desc = "Reverse Primer 4" (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 17: TGGTATGCAT AAGCTTGAAT TCAGGTAAAC AAGATATAAT TT 42 (2) INFORMATION FOR SEQ ID NO: 18: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 32 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (A) DESCRIPTION: /desc = "HLIP-A Primer 5" (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 18: CCCATTTAAA TATGAGGAGC TCCCTTGTGC TG 32 (2) INFORMATION FOR SEQ ID NO: 19: SUBSTITUTE SHEET (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 32 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (A) DESCRIPTION: /desc = "HLIP-B Primer 6" (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 19: CCCTTAATTA ACTAAAGACA TGTCCCAATT AA 32 (2) INFORMATION FOR SEQ ID NO: 20: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 6 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide addition (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 20: Arg-Pro-Val-Ser-Gln-Asp 5 (2) INFORMATION FOR SEQ ID NO: 21: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 5 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide addition (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 21: Ser-Pro-Ile-Arg-Met 5 (2) INFORMATION FOR SEQ ID NO: 22: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 6 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide addition (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 22: Ser-Pro-Ile-Arg-Ala-Arg 5 (2) INFORMATION FOR SEQ ID NO: 23: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 6 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide addition (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 23: Ser-Pro-Ile-Arg-Pro-Arg 5 (2) INFORMATION FOR SEQ ID NO: 24: (i) SEQUENCE CHARACTERISTICS: TTTUTE SHEET (A) LENGTH: 6 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide addition (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 24: Ser-Pro-Ile-Arg-Glu-Arg 5 (2) INFORMATION FOR SEQ ID NO: 25: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 5 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide addition (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 25: Ser-Pro-Ile-Arg-Lys 5 (2) INFORMATION FOR SEQ ID NO: 26: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 5 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide addition (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 26: Ser-Pro-Ile-Lys-Lys 5 (2) INFORMATION FOR SEQ ID NO: 27: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 6 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide addition (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 27: Ser-Pro-Ile-Arg-Arg-Pro 5 (2) INFORMATION FOR SEQ ID NO: 28: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 5 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide addition (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 28: Ser-Pro-Pro-Arg-Arg 5 (2) INFORMATION FOR SEQ ID NO: 29: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 5 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear SUBSTTTUTE SHEET (ii) MOLECULE TYPE: peptide addition (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 29: Ser-Pro-Iso-Pro-A g 5 5 (2) INFORMATION FOR SEQ ID NO: 30: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 5 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide addition ( i) SEQUENCE DESCRIPTION: SEQ ID NO: 30: Ser-Pro-Arg-Pro-Arg 5 (2) INFORMATION FOR SEQ ID NO: 31: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide addition (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 31: Ser-Pro-Ile-Arg (2) INFORMATION FOR SEQ ID NO: 32: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 5 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide addition ( i) SEQUENCE DESCRIPTION: SEQ ID NO: 32: Ser-Pro-Ile-Arg-Arg 5 (2) INFORMATION FOR SEQ ID NO: 33: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide addition ( i) SEQUENCE DESCRIPTION: SEQ ID NO: 33: Ser-Cys-Ile-Arg-Arg 5 (2) INFORMATION FOR SEQ ID NO: 34: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 7 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide addition (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 34: Γ Ser-Pro-Ile-Arg-Pro-Arg-Pro 5 (2) INFORMATION FOR SEQ ID NO: 35: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 7 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide addition (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 35: Ser-Cys-Ile-Arg-Pro-Arg-Pro 5 (2) INFORMATION FOR SEQ ID NO: 36: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 7 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide addition (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 36: Ser-Pro-Arg-Arg-Pro-Arg-Thr 5 (2) INFORMATION FOR SEQ ID NO: 37: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 7 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide addition (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 37: Ser-Pro-Phe-Arg-Pro-Lys-Leu 5 (2) INFORMATION FOR SEQ ID NO: 38: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 6 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide addition (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 38: Ser-Pro-Pro-Arg-Arg-Pro 5 (2) INFORMATION FOR SEQ ID NO: 39: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 6 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide addition (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 39: Ser-Pro-Ile-Arg-Arg-Glu 5 SUBSTTTUTE SHEET (2) INFORMATION FOR SEQ ID NO: 40: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 6 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide addition (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 40: Ser-Pro-Pro-Arg-Pro-Pro 5 (2) INFORMATION FOR SEQ ID NO: 41: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 6 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide addition (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 41: Ser-Pro-Pro-Arg-Pro-Arg 5 (2) INFORMATION FOR SEQ ID NO: 42: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 6 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide addition (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 42: Ser-Pro-Pro-Trp-Trp-Pro 5 (2) INFORMATION FOR SEQ ID NO: 43: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 6 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide addition (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 43: Ser-Pro-Pro-Trp-Arg-Pro 5 (2) INFORMATION FOR SEQ ID NO: 44: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 6 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide addition (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 44: Ser-Pro-Pro-Arg-Trp-Pro 5 (2) INFORMATION FOR SEQ ID NO: 45: (i) SEQUENCE CHARACTERISTICS: SUBSTITUTE SHEET (A) LENGTH: 6 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide addition (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 45: Ser-Pro-Pro-Arg-Trp-Pro 5 (2) INFORMATION FOR SEQ ID NO: 46: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 6 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide addition (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 46: Ser-His-Trp-Arg-Arg-Trp 5 (2) INFORMATION FOR SEQ ID NO: 47: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 5 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide addition (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 47: Ser-His-Trp-Arg-Lys 5 (2) INFORMATION FOR SEQ ID NO: 48: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 5 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide addition (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 48: Ser-His-Trp-Arg-Arg 5 (2) INFORMATION FOR SEQ ID NO: 49: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 7 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide addition (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 49: Thr-Ala-Ile-Arg-Pro-Arg-Lys 5 (2) INFORMATION FOR SEQ ID NO: 50: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 7 amino acids (B) TYPE: amino acid TE SHEET (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide addition (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 50: Ser-Thr-Arg-Arg-Pro-Arg-Pro 7 5 (2) INFORMATION FOR SEQ ID NO: 51: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 7 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide addition (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 51: Gly-Pro-Ile-Arg ' -Pro-Arg-Pro 7 5 (2) INFORMATION FOR SEQ ID NO: 52: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 7 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide addition (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 52: Leu-Pro-Phe-Arg-Glu-Arg-Pro 5 (2) INFORMATION FOR SEQ ID NO: 53: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 7 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide addition (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 53: Ser-Arg-Ser-Arg-His-Asp-Ala 5 (2) INFORMATION FOR SEQ ID NO: 54: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 7 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide addition (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 54: Ile-Pro-Ile-Arg-Pro-Arg-Arg 5 (2) INFORMATION FOR SEQ ID NO: 55: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 7 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide addition (Xi ) SEQUENCE DESCRIPTION: SEQ ID NO: 55 : Ser-Thr-Arg-Arg-Pro-Arg-Pro 5 (2) INFORMATION FOR SEQ ID NO: 56: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 7 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide addition (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 56: Thr-Ala-Ile-Arg-Pro-Arg-Lys . 5 (2) INFORMATION FOR SEQ ID NO: 57: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 6 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide addition (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 57: Trp-Arg-Trp-Arg-Trp-Arg 5 (2) INFORMATION FOR SEQ ID NO: 58: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 5 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide addition (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 58: Glu-Pro-Ile-Arg-Arg 5 (2) INFORMATION FOR SEQ ID NO: 59: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 5 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide addition (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 59: Ser-His-Trp-Glu-Glu 5 (2) INFORMATION FOR SEQ ID NO: 60: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 8 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide addition (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 60: Arg-Pro-Arg-Pro-Arg-Pro-Arg-Pro 8 5 (2) INFORMATION FOR SEQ ID NO: 61: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 11 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide addition (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 61: Ser-Ser-Thr-Arg-Arg-Ala-Ser-Pro-Ile-Lys-Lys 11 5 10 (2) INFORMATION FOR SEQ ID NO: 62: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 11 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide addition (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 62: Ala-Trp-Trp-Pro-Ser-Pro-Ile-Arg-Pro-Arg-Pro 11 5 10 (2) INFORMATION FOR SEQ ID NO: 63: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 12 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide addition (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 63: Ala-Pro-Pro-Pro-Arg-Pro-Arg-Pro-Arg-Pro-Arg-Pro 12 5 10 (2) INFORMATION FOR SEQ ID NO: 64: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 12 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide addition (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 64: Ala-Pro-Pro-Pro-Arg-Thr-Arg-Pro-Arg-Pro-Arg-Ser 12 5 10 (2) INFORMATION FOR SEQ ID NO: 65: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 8 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide addition (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 65: SHEET Ser-Pro-Lys-Arg-Lys-Pro-Arg-Pro 5 (2) INFORMATION FOR SEQ ID NO: 66: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 9 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide addition (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 66: Ser-Gln-Arg-Ile-Lys-Gln-Arg-Ile-Lys 5 (2) INFORMATION FOR SEQ ID NO: 67: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 8 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide addition (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 67: Ser-Pro-Pro-Pro-Arg-Pro-Arg-Pro 5 (2) INFORMATION FOR SEQ ID NO: 68: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 10 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide addition (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 68: Ser-Pro-Ile-Arg-Pro-Arg-Pro-Arg-Pro-Arg 10 5 10 (2) INFORMATION FOR SEQ ID NO: 69: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 9 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide addition (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 69: Ser-Pro-Ile-Arg- ys-Ala-Trp-Trp-Pro 5 (2) INFORMATION FOR SEQ ID NO: 70: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 12 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide addition (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 70: Ala-Pro-Pro-Pro-Lys-Ala-Ser-Pro-Arg-Gln-Arg-Pro 12 5 10 (2) INFORMATION FOR SEQ ID NO:71: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 15 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide addition ( i) SEQUENCE DESCRIPTION: SEQ ID NO: 71: Ser-Pro-Ile-Arg-Pro-Arg-Pro-Ser-Pro-Ile-Arg-Pro-Arg-Pro-Arg 15 5 10 15 (2) INFORMATION FOR SEQ ID NO: 72: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 8 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide addition (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 72: Ser-Pro-Pro-Arg-Trp-Pro-Arg-Arg 5 (2) INFORMATION FOR SEQ ID NO: 73: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 8 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide addition (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 73: Ser-Pro-Pro-Arg-Trp-Pro-Arg-Trp 5 (2) INFORMATION FOR SEQ ID NO: 74: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 8 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide addition (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 74: Ser-Pro-Pro-Arg-Trp-Pro-Trp-Arg 5 (2) INFORMATION FOR SEQ ID NO: 75: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 8 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide addition (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 75: Ser-Pro-Pro-Trp-Arg-Pro-Arg-Arg 5 (2) INFORMATION FOR SEQ ID NO: 76: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 8 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide addition (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 76: Ser-Pro-Pro-Trp-Trp-Pro-Arg-Trp 5 (2) INFORMATION FOR SEQ ID NO: 77: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 8 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide addition (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 77: Ser-Pro-Pro-Trp-Trp-Pro-Trp-Arg 5 (2) INFORMATION FOR SEQ ID NO: 78: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 8 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide addition (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 78: Ser-Pro-Pro-Trp-Trp-Pro-Trp-Trp 5 (2) INFORMATION FOR SEQ ID NO: 79: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 8 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide addition (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 79: Ser-Pro-Pro-Trp-Pro-Arg-Pro-Arg-Pro 5 (2) INFORMATION FOR SEQ ID NO: 80: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 7 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide addition (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 80: Ser -Ser- Lys-Gln- Asp- Tyr- Arg 5 (2) INFORMATION FOR SEQ ID NO: 81: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 42 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (A) DESCRIPTION: /desc = "Primer TiK 57" (xi) SEQUENCE DESCRIPTION: SEQ ID No. 81: CTTGGCTAGC CCTATACGTA GATCATCCAC ACAAGATTAT CG 42 (2) INFORMATION FOR SEQ ID NO: 82: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 32 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (A) DESCRIPTION: /desc = "Primer TiK 58" (xi) SEQUENCE DESCRIPTION: SEQ ID No. 82: CTTGGCTAGC TCCACACAAG ATTATCGTAT TG 32 (2) INFORMATION FOR SEQ ID NO: 83: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 32 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (A) DESCRIPTION: /desc = "Primer TiK 59" (xi) SEQUENCE DESCRIPTION: SEQ ID No. 83: GCCCTCTAGA CTATAAACAG AGACCAGTGT TC 32 (2) INFORMATION FOR SEQ ID NO: 84: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 83 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (A) DESCRIPTION: /desc = "Primer TiK 60" ( B) 5: 90% A and 3.33% of each C, G, T. 6: 90% C and 3.33% of each A, G, T. 7: 90% G and 3.33% of each A, C, T. 8: 90% T and 3.33% of each A, C, G. (xi) SEQUENCE DESCRIPTION: SEQ ID No. 84: GTAGTTTTTC GTGGT5CA57 (C/G)TCA58 (5/8) 67 (C/G) 556(7/8) (6/7) (C/G) 58 (5/8) 7 6 (T/G) 75658 (5/8) 78 (T/G) 88878 (T/G) 66 (G/C) 78 (C/G) 55885TC CACCTGTTAA TGG 83 (2) INFORMATION FOR SEQ ID NO: 85: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 17 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (A) DESCRIPTION: /desc = "Primer TiK 61" (xi) SEQUENCE DESCRIPTION: SEQ ID No. 85: AGAACAGCTG TTGCACC 17 (2) INFORMATION FOR SEQ ID NO: 86: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (A) DESCRIPTION: /desc = "Primer TiK 62" (xi) SEQUENCE DESCRIPTION: SEQ ID No. 86: CCGGGGA TCCACCATG 19 (2) INFORMATION FOR SEQ ID NO: 87: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 20 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (A) DESCRIPTION: /desc = "Primer TiK 64" (xi) SEQUENCE DESCRIPTION: SEQ ID No. 87: GCCCTCTAGA CTATAAACAG 20 (2) INFORMATION FOR SEQ ID NO: 88: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 17 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (A) DESCRIPTION: /desc = "Primer TiK 66" (xi) SEQUENCE DESCRIPTION: SEQ ID No. 88: CTGCAGAACT GTCATTC 17 (2) INFORMATION FOR SEQ ID NO: 89: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 16 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (A) DESCRIPTION: /desc = "Primer TiK 72" (xi) SEQUENCE DESCRIPTION: SEQ ID No. 89: TTGAGCTTGT ACCACG 16 (2) INFORMATION FOR SEQ ID NO: 90: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 36 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (A) DESCRIPTION: /desc = "Primer TiK 74" (xi) SEQUENCE DESCRIPTION: SEQ ID No. 90: CCGGGGATCC ACCATGAGAT TTCCTTCTAT TTTTAC 36 (2) INFORMATION FOR SEQ ID NO: 91: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 31 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (A) DESCRIPTION: /desc = "Primer TiK 75" (xi) SEQUENCE DESCRIPTION: SEQ ID No. 91: TGGAGCTAGC TCTTTTATCC AAAGAAACAC C 31 (2) INFORMATION FOR SEQ ID NO: 92: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 85 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: other nucleic acid (A) DESCRIPTION: /desc = "Primer TiK 76" (B) 5: 90% G and 3.33% of each C, G, T. 6: 90% A and 3.33% of each A, G, T. 7: 90% T and 3.33% of each A, C, T. 8: 90% C and 3.33% of each A, C, G. (xi) SEQUENCE DESCRIPTION: SEQ ID No. 92: CAGCCAATGC ATACTGC655 6885776778 8755755765 575565875T 88786775T5 57577GCATC CAATTTGCAA ATTAC 85 (2) INFORMATION FOR SEQ ID NO: 93: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 1115 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (vi) ORIGINAL SOURCE: (B) STRAIN: Absidia reflexa ATTC 44896 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 93: AAAGGCATTC TCATTTTGTA GTCTTATTGC TAGCAGTATT CATCTGCATG TGCTCTGTAT 60 CGGGTGTGCC ACTGCAAATT GATCCACGCG ATGACAAGAG CTATGTTCCT GAACAATATC 120 CTTTGAAGGT GAATGGTCCT TTGCCAGAAG GTGTAAGCGT GATCCAAGGC TATTGTGAAA 180 ACTGTACCAT GTATCCTGAA AAAAATAGTG TATCGGCATT CTCGTCATCA TCCACACAAG 240 ATTATCGTAT TGCAAGCGAG GCAGAGATTA AGGCACACAC ATTTTACACA GCATTGTCAG 300 CCAATGCATA CTGCAGAACT GTCATTCCTG GTGGTCGATG GAGCTGTCCC CACTGTGGTG 360 TTGCATCCAA TTTGCAAATT ACCAAGACTT TCAGCACCTT AATCACTGAT ACTAATGTCT 420 TGGTGGCTGT TGGCGAAAAG GAGAAGACCA TCTATGTAGT TTTTCGTGGT ACAAGCTCAA 480 TTCGCAACGC CATTGCTGAC ATTGTTTTTG TACCAGTGAA TTATCCACCT GTTAATGGAG 540 CCAAAGTACA CAAAGGATTT CTTGATAGCT ATAACGAAGT CCAGGATAAA CTTGTTGCTG 600 AAGTCAAGGC ACAACTTGAT CGTCATCCAG GATACAAGAT CGTCGTCACT GGACATTCCT 660 TGGGAGGTGC AACAGCTGTT CTCAGTGCAC TTGACCTTTA TCACCATGGC CATGCCAATA 720 TCGAAATCTA TACTCAAGGT CAGCCACGTA TAGGTACTCC AGCATTTGCA AACTATGTGA 780 TAGGCACCAA GATTCCATAC CAACGTCTTG TCCATGAGCG TGACATTGTT CCTCACCTTC 840 CACCTGGTGC ATTTGGTTTC TTGCATGCTG GTGAAGAGTT TTGGATCATG AAAGATAGCT 900 CGTTGCGCGT ATGTCCAAAT GGCATTGAAA CTGACAACTG CAGCAACTCC ATTGTTCCCT 960 TCACTAGTGT CATTGACCAT TTAAGCTATC TTGACATGAA CACTGGTCTC TGTTTATAAT 1020 CTTTAGTATC ATCCACTCCT CCTCTTTAAT GCAATACTTT TTAAGATAAA TCACAAGTAT 1080 ACTTTGTACA AAACCAAAAA AAAAAAAAAA AAAAA 1115 (2) INFORMATION FOR SEQ ID NO: 94: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 1020 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: circular (ii) MOLECULE TYPE: other nucleic acid (A) DESCRIPTION: /desc = "Yeast expression vector pTik05" (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION:3..1020 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 94: CA CAT ACA GGA ATT CAT TCA AGA ATA GTT CAA ACA AGA AGA TTA CAA 47 His Thr Gly Ile His Ser Arg Ile Val Gin Thr Arg Arg Leu Gin 1 5 10 15 ACT ATC AAT TTC ATA CAC AAT ATA AAC GAC GGT ACC CGG GGA TCC ACC 95 Thr Ile Asn Phe Ile His Asn Ile Asn Asp Gly Thr Arg Gly Ser Thr 20 25 30 ATG AGG AGC TCC CTT GTG CTG TTC TTT GTC TCT GCG TGG ACG GCC TTG 143 Met Arg Ser Ser Leu Val Leu Phe Phe Val Ser Ala Trp Thr Ala Leu 35 40 45 GCT AGC TCC ACA CAA GAT TAT CGT ATT GCA AGC GAG GCA GAG ATT AAG 191 Ala Ser Ser Thr Gin Asp Tyr Arg Ile Ala Ser Glu Ala Glu Ile Lys 50 55 60 GCA CAC ACA TTT TAC ACA GCA TTG TCA GCC AAT GCA TAC TGC AGA ACT . 239 Ala His Thr Phe Tyr Thr Ala Leu Ser Ala Asn Ala Tyr Cys Arg Thr 65 70 75 GTC ATT CCT GGT GGT CGA TGG AGC TGT CCC CAC TGT GGT GTT GCA TCC 287 Val Ile Pro Gly Gly Arg Trp Ser Cys Pro His Cys Gly Val Ala Ser 80 85 90 95 AAT TTG CAA ATT ACC AAG ACT TTC AGC ACC TTA ATC ACT GAT ACT AAT 335 Asn Leu Gin Ile Thr Lys Thr Phe Ser Thr Leu Ile Thr Asp Thr Asn 100 105 110 GTC TTG GTG GCT GTT GGC GAA AAG GTT GTT TTT GTA CCA GTG AAT TAT 383 Val Leu Val Ala Val Gly Glu Lys Val Val Phe Val Pro Val Asn Tyr 115 120 125 CCA CCT GTT AAT GGA GCC AAA GTA CAC AAA GGA TTT CTT GAT AGC TAT 431 Pro Pro Val Asn Gly Ala Lys Val His Lys Gly Phe Leu Asp Ser Tyr 130 135 140 AAC GAA GTC CAG GAT AAA CTT GTT GCT GAA GTC AAG GCA CAA CTT GAT 479 Asn Glu Val Gin Asp Lys Leu Val Ala Glu Val Lys Ala Gin Leu Asp 145 150 155 CGT CAT CCA GGA TAC AAG ATC GTC GTC ACT GGA CAT TCC TTG GGA GGT 527 Arg His Pro Gly Tyr Lys Ile Val Val Thr Gly His Ser Leu Gly Gly 160 165 170 175 GCA ACA GCT GTT CTC AGT GCA CTT GAC CTT TAT CAC CAT GGC CAT GCC 575 Ala Thr Ala Val Leu Ser Ala Leu Asp Leu Tyr His His Gly His Ala 180 185 190 AAT ATC GAA ATC TAT ACT CAA GGT CAG CCA CGT ATA GGT ACT CCA GCA 623 Asn Ile Glu Ile Tyr Thr Gin Gly Gin Pro Arg Ile Gly Thr Pro Ala 195 200 205 TTT GCA AAC TAT GTG ATT GGC ACC AAG ATT CCA TAC CAA CGT CTT GTC 671 Phe Ala Asn Tyr Val Ile Gly Thr Lys Ile Pro Tyr Gin Arg Leu Val 210 215 220 CAT GAG CGT GAC ATT GTT CCT CAC CTT CCA CCT GGT GCA TTT GGT TTC 719 His Glu Arg Asp Ile Val Pro His Leu Pro Pro Gly Ala Phe Gly Phe 225 230 235 TTG CAT GCT GGT GAA GAG TTT TGG ATC ATG AAA GAT AGC TCG TTG CGC 767 Leu His Ala Gly Glu Glu Phe Trp Ile Met Lys Asp Ser Ser Leu Arg 240 245 250 255 GTA TGT CCA AAT GGC ATT GAA ACT GAC AAC TGC AGC AAC TCC ATT GTT 815 Val Cys Pro Asn Gly Ile Glu Thr Asp Asn Cys Ser Asn Ser Ile Val 260 265 270 CCC TTC ACT AGT GTC ATT GAC CAT TTA AGC TAT CTT GAC ATG AAC ACT 863 Pro Phe Thr Ser Val Ile Asp His Leu Ser Tyr Leu Asp Met Asn Thr 275 280 285 GGT CTC TGT TTA TAG TCT AGA GGG CCG CAT GAT GTA ATT AGT TAT GTC 911 Gly Leu Cys Leu * Ser Arg Gly Pro His Asp Val Ile Ser Tyr Val 290 295 300 ACG CTT ACA TTC ACG CCC TCC CCC CAC ATC CGC TCT AAC CGA AAA GGA 959 Thr Leu Thr Phe Thr Pro Ser Pro His Ile Arg Ser Asn Arg Lys Gly 305 310 315 AGG AGT TAG ACA ACC TGA AGT CTA GGT CCC TAT TTA TTT TTT TAT AGT 1007 Arg Ser * Thr Thr * Ser Leu Gly Pro Tyr Leu Phe Phe Tyr Ser 320 325 330 335 TAT GTT AGT ATT A 1020 Tyr Val Ser Ile (2) INFORMATION FOR SEQ ID NO: 95: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 339 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 95: His Thr Gly Ile His Ser Arg Ile Val Gin Thr Arg Arg Leu Gin Thr 1 5 10 15 Ile Asn Phe Ile His Asn Ile Asn Asp Gly Thr Arg Gly Ser Thr Met 20 25 30 Arg Ser Ser Leu Val Leu Phe Phe Val Ser Ala Trp Thr Ala Leu Ala 35 40 45 Ser Ser Thr Gin Asp Tyr Arg Ile Ala Ser Glu Ala Glu Ile Lys Ala 50 55 60 His Thr Phe Tyr Thr Ala Leu Ser Ala Asn Ala Tyr Cys Arg Thr Val 65 70 75 80 Ile Pro Gly Gly Arg Trp Ser Cys Pro His Cys Gly Val Ala Ser Asn 85 90 95 Leu Gin Ile Thr Lys Thr Phe Ser Thr Leu lie Thr Asp Thr Asn Val 100 105 110 Leu Val Ala Val Gly Glu Lys Val Val Phe Val Pro Val Asn Tyr Pro 115 120 125 Pro Val Asn Gly Ala Lys Val His Lys Gly Phe Leu Asp Ser Tyr Asn 130 135 140 Glu Val Gin Asp Lys Leu Val Ala Glu Val Lys Ala Gin Leu Asp Arg 145 150 155 160 His Pro Gly Tyr Lys Ile Val Val Thr Gly His Ser Leu Gly Gly Ala 165 170 175 Thr Ala Val Leu Ser Ala Leu Asp Leu Tyr His His Gly His Ala Asn 180 185 190 Ile Glu Ile Tyr Thr Gin Gly Gin Pro Arg Ile Gly Thr Pro Ala Phe 195 200 205 Ala Asn Tyr Val Ile Gly Thr Lys Ile Pro Tyr Gin Arg Leu Val His 210 215 220 Glu Arg Asp Ile Val Pro His Leu Pro Pro Gly Ala Phe Gly Phe Leu 225 230 235 240 His Ala Gly Glu Glu Phe Trp Ile Met Lys Asp Ser Ser Leu Arg Val 245 250 255 Cys Pro Asn Gly Ile Glu Thr Asp Asn Cys Ser Asn Ser Ile Val Pro 260 265 270 Phe Thr Ser Val Ile Asp His Leu Ser Tyr Leu Asp Met Asn Thr Gly 275 280 285 Leu Cys Leu * Ser Arg Gly Pro His Asp Val Ile Ser Tyr Val Thr 290 295 300 Leu Thr Phe Thr Pro Ser Pro His Ile Arg Ser Asn Arg Lys Gly Arg 305 310 315 320 Ser * Thr Thr * Ser Leu Gly Pro Tyr Leu Phe Phe Tyr Ser Tyr 325 330 335 Val Ser Ile (2) INFORMATION FOR SEQ ID NO: 96: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 255 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (vi) ORIGINAL SOURCE: (A) ORGANISM: MFαl signal sequence (mating factor) (B) STRAIN: yeast strain YPH499 (ix) FEATURE: (A) NAME/KEY: sig_peptide (B) LOCATION: 1..255 (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 1..255 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 96: ATG AGA TTT CCT TCT ATT TTT ACT GCT GTT TTA TTC GCT GCT TCC TCC 48 Met Arg Phe Pro Ser Ile Phe Thr Ala Val Leu Phe Ala Ala Ser Ser 1 5 10 15 GCT TTA GCT GCT CCA GTC AAC ACT ACC ACT GAA GAT GAA ACG GCT CAA 96 Ala Leu Ala Ala Pro Val Asn Thr Thr Thr Glu Asp Glu Thr Ala Gin 20 25 30 ATT CCA GCT GAA GCT GTC ATC GGT TAC CTT GAT TTA GAA GGT GAT TTC 144 Ile Pro Ala Glu Ala Val Ile Gly Tyr Leu Asp Leu Glu Gly Asp Phe 35 40 45 GAT GTT GCT GTT TTG CCA TTT TCC AAC TCC ACC AAT AAC GGT TTA TTG 192 Asp Val Ala Val Leu Pro Phe Ser Asn Ser Thr Asn Asn Gly Leu Leu 50 55 60 TTT ATC AAT ACT ACT ATT GCC TCC ATT GCT GCT AAA GAA GAA GGT GTT 240 Phe Ile Asn Thr Thr Ile Ala Ser Ile Ala Ala Lys Glu Glu Gly Val 65 70 75 80 TCT TTG GAT AAA AGA 255 Ser Leu Asp Lys Arg 85 (2) INFORMATION FOR SEQ ID NO: 97: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 85 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 97: Met Arg Phe Pro Ser Ile Phe Thr Ala Val Leu Phe Ala Ala Ser Ser 1 5 10 15 Ala Leu Ala Ala Pro Val Asn Thr Thr Thr Glu Asp Glu Thr Ala Gin 20 25 30 Ile Pro Ala Glu Ala Val Ile Gly Tyr Leu Asp Leu Glu Gly Asp Phe 35 " 40 45 Asp Val Ala Val Leu Pro Phe Ser Asn Ser Thr Asn Asn Gly Leu Leu 50 55 60 Phe Ile Asn Thr Thr Ile Ala Ser Ile Ala Ala Lys Glu Glu Gly Val 65 70 75 80 Ser Leu Asp Lys Arg 85 (2) INFORMATION FOR SEQ ID NO: 98: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein ( i) SEQUENCE DESCRIPTION: SEQ ID NO: 98: MRFYSWSLLAVSICTYGVSGVPVQIGPRDKSYVPEQ YPLKMNGPLPEGVSVIQGYCENCTMYPEENS- VTALSSSKQDYRTASETEIQAHTFYTALSANAYCRNVIPGGRWSCPHCDVTSNLKITKTFSTLITDTNVAVAV GEKEKTIYIVFRGTNSIRNAIADIVFVPVDYPPVDGAKVHKGFLDSYNEVQDQLVAEVKKQLDNHPGYKIWA GHSLGGATAVLCALDLYHHGHHNIEIYTQGQPRVGTPAFAKYVIGTKIPYQRLVNERDIVPHLPPGAFGFLHA GEEFWIMKDSSLRVCPNGIETDDCSNSIVPFTSVIDHLSYLDMNTGLCL* (2) INFORMATION FOR SEQ ID NO: 99: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 864 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (vi) ORIGINAL SOURCE: (B) STRAIN: Pseudomonas sp. (ix) FEATURE: (A) NAME/KEY: mat_peptide (B) LOCATION: 1..864 (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 1..864 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 99: TTC GGC TCC TCG AAC TAC ACC AAG ACC CAG TAC CCG ATC GTC CTG ACC 48 Phe Gly Ser Ser Asn Tyr Thr Lys Thr Gin Tyr Pro Ile Val Leu Thr 1 5 10 15 CAC GGC ATG CTC GGT TTC GAC AGC CTG CTT GGA GTC GAC TAC TGG TAC 96 His Gly Met Leu Gly Phe Asp Ser Leu Leu Gly Val Asp Tyr Trp Tyr 20 25 30 GGC ATT CCC TCA GCC CTG CGT AAA GAC GGC GCC ACC GTC TAC GTC ACC 144 Gly Ile Pro Ser Ala Leu Arg Lys Asp Gly Ala Thr Val Tyr Val Thr 35 40 45 GAA GTC AGC CAG CTC GAC ACC TCC GAA GCC CGA GGT GAG CAA CTG CTG 192 Glu Val Ser Gin Leu Asp Thr Ser Glu Ala Arg Gly Glu Gin Leu Leu 50 55 60 ACC CAA GTC GAG GAA ATC GTG GCC ATC AGC GGC AAG CCC AAG GTC AAC 240 Thr Gin Val Glu Glu Ile Val Ala Ile Ser Gly Lys Pro Lys Val Asn 65 70 75 80 CTG TTC GGC CAC AGC CAT GGC GGG CCT ACC ATC CGC TAC GTT GCC GCC 288 Leu Phe Gly His Ser His Gly Gly Pro Thr Ile Arg Tyr Val Ala Ala 85 90 95 GTG CGC CCG GAT CTG GTC GCC TCG GTC ACC AGC ATT GGC GCG CCG CAC 336 Val Arg Pro Asp Leu Val Ala Ser Val Thr Ser Ile Gly Ala Pro His 100 105 110 AAG GGT TCG GCC ACC GCC GAC TTC ATC CGC CAG GTG CCG GAA GGA TCG 384 Lys Gly Ser Ala Thr Ala Asp Phe Ile Arg Gin Val Pro Glu Gly Ser 115 120 125 GCC AGC GAA GCG ATT CTG GCC GGG ATC GTC AAT GGT CTG GGT GCG CTG 432 Ala Ser Glu Ala Ile Leu Ala Gly Ile Val Asn Gly Leu Gly Ala Leu 130 135 140 ATC AAC TTC CTT TCC GGC AGC AGT TCG GAC ACC CCA CAG AAC TCG CTG 480 Ile Asn Phe Leu Ser Gly Ser Ser Ser Asp Thr Pro Gin Asn Ser Leu 145 150 155 160 GGC ACG CTG GAG TCA CTG AAC TCC GAA GGC GCC GCA CGG TTT AAC GCC 528 Gly Thr Leu Glu Ser Leu Asn Ser Glu Gly Ala Ala Arg Phe Asn Ala 165 170 175 CGC TTC CCC CAG GGG GTA CCA ACC AGC GCC TGC GGC GAG GGC GAT TAC 576 Arg Phe Pro Gin Gly Val Pro Thr Ser Ala Cys Gly Glu Gly Asp Tyr 180 185 190 GTG GTC AAT GGC GTG CGC TAT TAC TCC TGG AGG GGC ACC AGC CCG CTG 624 Val Val Asn Gly Val Arg Tyr Tyr Ser Trp Arg Gly Thr Ser Pro Leu 195 200 205 ACC AAC GTA CTC GAC CCC TCC GAC CTG CTG CTC GGC GCC ACC TCC CTG 672 Thr Asn Val Leu Asp Pro Ser Asp Leu Leu Leu Gly Ala Thr Ser Leu 210 215 220 ACC TTC GGT TTC GAG GCC AAC GAT GGT CTG GTC GGA CGC TGC AGC TCC 720 Thr Phe Gly Phe Glu Ala Asn Asp Gly Leu Val Gly Arg Cys Ser Ser 225 230 235 240 CGG CTG GGT ATG GTG ATC CGC GAC AAC TAC CGG ATG AAC CAC CTG GAC 768 Arg Leu Gly Met Val Ile Arg Asp Asn Tyr Arg Met Asn His Leu Asp 245 250 255 GAG GTG AAC CAG ACC TTC GGG CTG ACC AGC ATC TTC GAG ACC AGC CCG 816 Glu Val Asn Gin Thr Phe Gly Leu Thr Ser Ile Phe Glu Thr Ser Pro 260 265 270 GTA TCG GTC TAT CGC CAG CAA GCC AAT CGC CTG AAG AAC GCC GGG CTC 864 Val Ser Val Tyr Arg Gin Gin Ala Asn Arg Leu Lys Asn Ala Gly Leu 275 280 285 (2) INFORMATION FOR SEQ ID NO: 100: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 288 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 100: Phe Gly Ser Ser Asn Tyr Thr Lys Thr Gin Tyr Pro Ile Val Leu Thr 1 5 10 15 His Gly Met Leu Gly Phe Asp Ser Leu Leu Gly Val Asp Tyr Trp Tyr 20 25 30 Gly Ile Pro Ser Ala Leu Arg Lys Asp Gly Ala Thr Val Tyr Val Thr 35 40 45 Glu Val Ser Gin Leu Asp Thr Ser Glu Ala Arg Gly Glu Gin Leu Leu 50 55 60 Thr Gin Val Glu Glu Ile Val Ala Ile Ser Gly Lys Pro Lys Val Asn 65 70 75 80 Leu Phe Gly His Ser His Gly Gly Pro Thr Ile Arg Tyr Val Ala Ala 85 90 95 Val Arg Pro Asp Leu Val Ala Ser Val Thr Ser Ile Gly Ala Pro His 100 105 110 Lys Gly Ser Ala Thr Ala Asp Phe Ile Arg Gin Val Pro Glu Gly Ser 115 120 125 Ala Ser Glu Ala Ile Leu Ala Gly Ile Val Asn Gly Leu Gly Ala Leu 130 135 140 Ile Asn Phe Leu Ser Gly Ser Ser Ser Asp Thr Pro Gin Asn Ser Leu 145 150 155 160 Gly Thr Leu Glu Ser Leu Asn Ser Glu Gly Ala Ala Arg Phe Asn Ala 165 170 175 Arg Phe Pro Gin Gly Val Pro Thr Ser Ala Cys Gly Glu Gly Asp Tyr 180 185 190 Val Val Asn Gly Val Arg Tyr Tyr Ser Trp Arg Gly Thr Ser Pro Leu 195 200 205 Thr Asn Val Leu Asp Pro Ser Asp Leu Leu Leu Gly Ala Thr Ser Leu 210 215 220 Thr Phe Gly Phe Glu Ala Asn Asp Gly Leu Val Gly Arg Cys Ser Ser 225 230 235 240 Arg Leu Gly Met Val Ile Arg Asp Asn Tyr Arg Met Asn His Leu Asp " 245 250 255 Glu Val Asn Gin Thr Phe Gly Leu Thr Ser Ile Phe Glu Thr Ser Pro 260 265 270 Val Ser Val Tyr Arg Gin Gin Ala Asn Arg Leu Lys Asn Ala Gly Leu 275 280 285

Claims: CLAIMS 1. A lipolytic enzyme which, when present in detergent composition A and/or B defined herein, is capable of removmg at least 15% more lard from a lard stained swatch than the same detergent composition without the enzyme, in a one cycle wash assay comprising 5 subjecting 7 lard-stained cotton swatches (9 x 9 cm) per beaker to a one cycle wash in a thermostated Terg-O-to-Meter (TOM), each beaker containing 1000 ml of water comprising 3.2 mM Ca 2+ /Mg 2+ (in a ratio of 5:1) and 5 g/1 of said detergent composition, pH 10, and comprising 12500 LU/1 of the Upolytic enzyme, the wash treatment being carried out for 20 minutes at a temperature of 30°C, followed by rinsing for 15 minutes in running tap water 0 and overnight linedrying at room temperature, subsequent extraction and quantification of fatty matter on the swatches by Soxhlet extraction. 2. The hpolytic enzyme according to claim 1, wherein at least 20% more lard is removed. 3. 3. The Upolytic enzyme according to claim 2, wherein at least 30 % more lard is 5 removed. 4. The Upolytic enzyme according to any of the preceding claims, which exhibits lipolytic activity towards mono-, di- and/or triglycerides and/or cutin. 5. The Upolytic enzyme according to any of claims 1-4, which is a variant of a parent Upolytic enzyme which comprises at least 1, such as at least 3 mutations as 0 compared to the parent enzyme. 6. The lipolytic enzyme according to claim 5, in which at least one neutral amino acid residue of the parent enzyme has been replaced with a positively charged amino acid residue, at least one negatively charged amino acid residue has been deleted, or at least one negatively charged amino acid residue has been replaced with a neutral or positively 5 charged amino acid residue. 7. The lipolytic enzyme according to claim 5, which, compared to a parent lipolytic enzyme, has been modified by deletion or substitution of at least one amino acid residue in a Upid contact zone of the parent lipolytic enzyme or addition of at least one amino acid residue to said lipid contact zone. 8. The lipolytic enzyme according to claim 7, which is a variant of a parent lipolytic enzyme which comprises at least one mutation within and at least one mutation outside the lipid contact zone of the parent lipolytic enzyme. 9. The lipolytic enzyme according to any of the preceding claims, which further comprises a peptide addition appUed at or within the C- and/or N-terminal end of the mature form of the parent enzyme. 10. The Upolytic enzyme according to any of claims 5-9, wherein the mutation and or the peptide addition is selected so as to increase the affinity of the parent enzyme towards its substrate. 11. The Upolytic Upolytic enzyme according to claim 9 or 10, wherein the peptide addition is one which is capable of forming a covalent binding to the mature part of the parent enzyme. 12. The Upolytic enzyme according to any of claims 9-11, which comprises a cysteine residue in the peptide addition and a cystein residue in the mature part of the parent enzyme in such a manner that the said cystein residues together forms a cystein bridge. 13. The Upolytic enzyme according to any of claims 9-12, wherein the peptide addition is one which has a low susceptibility to proteolytic degradation by proteolytic enzymes of a host ceU used for expressing the first wash Upolytic enzyme. 14. The Upolytic enzyme according to claim 13, wherein the peptide addition comprises at least one proline residue, such as two or three proline residues. 15. The Upolytic enzyme according to any of claims 9-14, wherein the peptide addition comprises at least one, such as one, two or three positive or hydrophobic amino acids resi¬ dues. 16. The Upolytic enzyme according to any of claims 9-15, wherein the length of the pep¬ tide addition is from 1 and 500 amino acids, preferably 1 to 200, more preferably 2 to 100, even more preferably 2 to 50, and most preferably between 1 and 15, such as 1 to 10 or 4 to 10 amino acids. 17. The Upolytic enzyme according to any of claims 9-16, wherein the peptide addition is one of the foUowing peptide additions: Arg (R), or Lys (K), or Leu (L), or ue (1), or Val (V), or Trp (W) or Phe (F), or Arg-Pro (RP), or Lys-Lys (KK), or Arg-Lys (RK), or Lys-Arg (KR), or Arg-Arg (RR), or Arg-Arg-Pro (RRP), or Arg-Pro-Val-Ser-Gln-Asp (RPVSQD) Ser-Pro-ϋe-Arg-Met (SPIRM), or Ser-Pro-ϋe-Arg-Ala-Arg (SPIRAR), or Ser-Pro-Ile-Arg-Pro-Arg (SPIRPR) or Ser-Pro-He-Arg-Giu-Arg (SPIRER), or Ser-Pro-ϋe-Arg-Lys (SPIRK), or Ser-Pro-ϋe-Lys-Lys (SPIKK), or Ser-Pro-Ile-Arg-Arg-Pro (SPIRRP), or Ser-Pro-Pro-Arg-Arg (SPPRR), or Ser-Pro-Iso-Pro-Arg (SPIPR), or Ser-Pro-Arg-Pro-Arg (SPRPR), or Ser-Pro-Ile-Arg (SPIR), or Ser-Pro-He-Arg-Arg (SPIRR), or Ser-Cys-ile-Arg-Arg, (SCIRR), or Ser-Pro-Ile-Arg-Pro-Arg-Pro (SPIRPRP), or Ser-Cys-πe-Arg-Prc-Arg-Pro (SCPIRPRP), or Ser-Pro-Arg-Arg-Pro-Arg-Thr (SPRRPRT), or Ser-Pro-Phe-Arg-Pro-Lys-Leu (SPFRPKL), or Ser-Pro-Pro-Arg-Arg-Pro (SPPRRP), or Ser-Pro-Ile-Arg-Arg-Glu (SPIRRE), or Ser-Pro-Pro-Arg-Pro-Pro (SPPRPP), or Ser-Pro-Pro-Arg-Pro-Arg (SPPRPR), or Ser-Pro-Pro-Trp-Trp-Pro (SPPWWP), or Ser-Pro-Pro-Trp-Arg-Pro (SPPWRP), or Ser-Pro-Pro-Arg-Trp-Pro (SPPRWP), or Ser-Pro-Pro-Arg-Trp-Pro (SPPRWP), or Ser-His-Trp-Arg-Arg-Trp (SHWRRW), or Ser-His-Trp-Arg-Lys (SHWRK), or Ser-His-Trp-Arg-Arg (SHWRR), or Thr-Ala-Ile-Arg-Pro-Arg-Lys (TAIRPRK), Ser-Thr-Arg-Arg-Pro-Arg-Pro (STRRPRP), Gly-Pro-Ηe-Arg-Pro-Arg-Pro (GPIRPRP), Leu-Pro-Phe-Arg-Gln-Arg-Pro (LPFRQRP), Ser-Arg-Ser-Arg-His-Asp-Ala (SRSRHNA), He-Pro-Ile-Arg-Pro-Arg-Arg (IPIRPRR), Ser-Thr-Arg-Arg-Pro-Arg-Pro (STRRPRP), Thr-Ala-Ile-Arg-Pro-Arg-Lys (TAIRPRK), Trp-Arg-Trp-Arg-Trp-Arg (WRWRWR), Gln-Pro-Ile-Arg-Arg (QPIRR), Ser-His-Trp-Gln-Gln (SHWQQ), Arg-Pro-Arg-Pro-Arg-Pro-Arg-Pro (RPRPRPRP), or Ser-Ser-Thr-Arg-Arg-Ala-Ser-Pro-Ηe-Lys-Lys (SSTRRASPIKK), or Ala-Trp-Trp-Pro-Ser-Pro-Ile-Arg-Pro-Arg-Pro (AWWPSPIRPRP), or Ala-Pro-Pro-Pro-Arg-Pro-Arg-Pro-Arg-Pro-Arg-Pro (APPPRPRPRPRP), or Ala-Pro-Pro-Prc-Arg-Thr-Arg-Pro-Arg-Pro-Arg-Ser (APPPRTRPRPRS), or Ser-Pro-Lys-Arg-Lys-Pro-Arg-Pro (SPKRKPRP), or Ser-Gln-Arg-πe-Lys-Gln-Arg-πe-Lys (SQRKQRIK), or Ser-Pro-Pro-Pro-Arg-Pro- Arg-Pro (SPPPRPRP), or Ser-Pro-Ile-Arg-Pro-Arg-Pro-Arg-Pro-Arg SPIRPRPRPR, or Ser-Pro-ne-Arg-Lys-Ala-Trp-Trp-Pro (SPIRKAWWP), or Ala-Pro-Pro-Pro-Lys-Ala-Ser-Pro-Arg-Gln-Arg-Pro (APPPKASPRQRP), or Ser-Pro-Ile-Arg-Pro-Arg-Pro-Ser-Pro-Ile-Arg-Pro-Arg-Pro-Arg (SPIRPRPSPIRPRP), or Ser-Pro-Pro-Arg-Trp-Pro-Arg-Arg (SPPRWPRR), or Ser-Pro-Pro-Arg-Trp-Pro-Arg-Trp (SPPRWPRW), or Ser-Pro-Pro-Arg-Trp-Pro-Trp-Arg (SPPRWPWR), or Ser-Pro-Pro-Trp-Arg-Pro-Arg-Arg (SPPWRPRR), or Ser-Pro-Pro-Trp-Trp-Pro-Arg-Trp (SPPWWPRW, or Ser-Pro-Pro-Trp-Trp-Pro-Tip-Arg (SPPWWPWR), or Ser-Pro-Pro-Trp-Trp-Pro-Trp-Tip (SPPWWPWW), or Trp-Pro (WP), or Ser-Pro-Pro-Trp-Pro-Arg-pro-Arg-Pro (SPPWPRPRP). 18. The Upolytic enzyme according to any of claims 9-17, which, in addition to the pep¬ tide addition, comprises a mutation in a non-structural part of the N-terminus and/or C- terminus of the parent enzyme, in particular a mutation which has resulted in the removal of at least one negatively charged amino acid residue. 19. The Upolytic enzyme according to claim 18, wherein a negatively charged amino acid residue of said non-structural part has been deleted or replaced by a neutral or positively charged amino acid residue or by a hydrophobic amino acid residue, or wherein a neu¬ tral amino acid residue has been replaced with a positively charged amino acid residue. 20. A Upolytic enzyme comprising a peptide addition according to any of claims 9-19, said peptide addition having been appUed by a) subjecting a DNA sequence encoding a parent enzyme with a peptide addition to ran¬ dom mutagenesis in the part of the DNA sequence encoding the peptide addition and optionaUy a non-structural N-terminal or C-terminal part of the mature form of the par- ent enzyme, b) expressing the resulting mutated DNA sequence in a suitable host ceU so as to produce a modified Upolytic enzyme, and c) screening for modified Upolytic enzymes resulting from step c) which has an improved performance as compared to the parent enzyme. 21. The Upolytic enzyme according to claim 20, wherein the random mutagenesis is per¬ formed so as to introduce one or more positively charged or hydrophobic amino acid resi¬ dues into the peptide addition and optionaUy the non-structural part of the parent enzyme. 22. The first wash lipolytic enzyme according to any of the preceding claims, which is derived from a microorganism. 23. The Upolytic enzyme according to claim 22, which is derived from a fungus or a bacterium. 24. The Upolytic enzyme according to claim 23, which is derived from a Humicola sp.or a Pseudomonas sp. 25. The Upolytic enzyme according to claim 24, which is derived from Humicola lanuginosa, in particular from the strain DSM 4109, or which is homologous to the Upolytic enzyme derived from the lipolytic enzyme derived from said organism. 26. The Upolytic enzyme according to claim 25, which comprises at least one mutation compared to the H. lanuginosa lipolytic enzyme having the amino acid sequence shown in SEQ ID No. 1. 27. The lipolytic enzyme according to claim 25 or 26, wherein one of the mutations is a deletion or substitution of an amino acid residue in a Upid contact zone of the Upolytic enzyme or an addition of an amino acid residue to the lipid contact zone. 28. The Upolytic enzyme according to claim 26, wherein one of the mutations is a deletion or substitution of an amino acid residue outside the lipid contact zone of the lipolytic enzyme or an addition of an amino acid residue to the part of the lipolytic enzyme located outside the lipid contact zone. 29. The lipolytic enzyme according to any of claims 26-28, in which is the result of a combination between at least two of the following mutants or a part thereof: (a) E56R,D57L,I90F,D96L,E99K (b) E56R,D57L,V60M,D62N,S83T,D96P,D102E (c) D57G,N94K,D96L,L97M (d) E87K,G91A,D96R,I100V,E129K,K237M,I252L,P256T,G263A,L264Q (e) E56R,D57G,S58F,D62C,T64R,E87G,G91A,F95L,D96P,K98I,(K237M) (f) E210K (g)9 S83T,N94K,D96N (h) E87K,D96V (i) N94K,D96A 0) E87K,G91A,D96A (k) D167G,E210V (1) S83T,G91A,Q249R (m) E87K,G91A (n) S83T,E87K,G91A,N94K,D96N,DlllN. (o) N73D,E87K,G91A,N94I,D96G. (p) L67P,I76V,S83T,E87N,I90N,G91A,D96A,K98R. (q) S83T,E87K,G91A,N92Η,N94K,D96M (s) S85P,E87K,G91A,D96L,L97V. (t) E87K,I90N,G91A,N94S,D96N,I100T. (U) I34V,S54P,F80L,S85T,D96G,R108W,G109V,D111G,S116P,L124S,V132M,V140Q,V1 41A,F142S,H145R,N162T,I166V,F181P,F183S,R205G,A243T,D254G,F262L. 30. The Upolytic enzyme according to any of claims 26-29, in which at least one of the following amino acid residues in SEQ ID No.1 has been replaced with another amino acid residue: A49, G59, S85, S116, Q126, D137, S170 or W221 31. The lipolytic enzyme according to any of claims 26- 30, which comprises at least one of the foUowing substitutions: A49P, G59V, I90F, S85F, S116P, Q126R, D137G, S170P or W221L or a substitution to an amino acid residue belonging to the same charge group as the indicated substituent. 32. The lipolytic enzyme according to claim 26, which comprises the following mutations: D57G+N94K+D96L+Q249R; D57G+N94K+D96L+S116P+Q249R, D57G+G59V+N94K+D96L+Q249R, D57G+N94K+D96L+S116P+S170P+Q249R; D57G+G59V+N94K+D96L+S170P+Q249R; D57G+N94K+D96L+S170P+Q249R; D167G+E210V+Q249R; E56K+D167G+E210V; D137G+D167G+E210V+Q249R; D167G+E210V+W221L+Q249R; D57G+N94K+F95L+D96H,L+Q249R; D57G+N94K+D96L+E210K; D57G+G59V+N94K+D96L+S116P+S170P+Q249R; S3R+D137G+D167G+E210V+W221L; D137G+D167G+E210V+W221L+N233R; S3R+I90F+D96L+E99K+V187A+Q249R; I90F+D96L+E99K+V187A+D233R; I90F+D96L+E99K+V187A+D234Y; I90F+D96L+E99K+V187A+T231R; I90F+D96L+E99K+V187A; D62R+I90F+D96L+E99K+V187A; I90F+D96L+E99K+V187A+N200R+R209A; I90F+D96L+E99K+V187A+T199R+N200R+R209A; D57G+D62R+N94K+D96L+Q249R; D57G+N94K+D96L+N200R+R209A-I-Q249R; D57G+N94K+D96L+T199R+N200R+Q249R; I90F+D96L+E99K+V187A+T199R; D57G+N94K+D96L+T199R+R209A+Q249R; I90F+D96L+E99K+V187A+Q249R; I90F+D96L+E99K+V187A+P253R; I90F+D96L+E99K+D137G+D167G+V187A+Q249R; I90F+D96L+E99K+D137G+V187A+Q249R; D96L+E99K+V187A+Q249R; V2P+N94K+D96L+Q249R; V2W+S3R+N94K+D96L+Q249R; V2R+S3R+N94K+D96L+Q249R; V2R+S3W+N94K+D96L+Q249R; V2W+S3R+N94K+D96L+Q249R; N94K+D96L+Q249R; V2G+S3T+D57G+N94K+D96L+L97M+Q249R; V2G+S3T+O4P+D5E+D57G+N94K+D96L+L97M+Q249R; V2G+D5Q+L6M +D57G+N94K+D96L+L97M+Q249R; D57G+G59V+N94K+D96L+L97M+S116P+S170P+Q249R; A49P+D167G+E210V; E56K+D57G+D62R+S83T+S85F+D96L+D102Y+E210K; D57G+N94K+D96L+L97M+Q249R; D137G+D167G+E210V+W221L; N94K+F95L+D96H+N101S+F181L+D234Y+I252L+P256T+G263A+L264Q; I90F+D96L+E99K+V187A; N94K+D96A+Q249R; A19P+D167G+E210V+W221L; N94K+D96L+L97M+Q249R; D57G+N94K+D96L+Q249R; I90F+D96L+E99K+D137G+V187A; N94K+D96L+E99K+Q249R; N94K+D96L+E99K+T231R+N233R+D234R+Q249R; N94K+D96L+E99K+D111N+F211 A+G225P+Q249R+T267R; or N94K+D96L+E99K+D111N+F211A+G225P+T231R+N233R+D234R+Q249R+ T267R. E1K+N94K+D96L+E99K+Q249R N94K+D96L+K223R+Q249R N94K+D96L+E99K+N233R N94K+D96L+E99K+T231R+N233R+Q249R N94K+D96L+E99K+N233R+Q249R N94K+D96L+E99K+D234R+Q249R 33. A Upolytic enzyme according to claim 32, which further comprises a peptide addition within or at its N-terminal and/or C-terminal end or which comprises a mutation in the non-structural part of the N-terminal and/or C-terminal end of the mature form of the enzyme. 34. A Upolytic enzyme according to claim 33, which comprises the foUowing mutations: SPIRPRP+D57G+N94K+D96L+Q249R; SPPRRP+I90F+D96L+E99K+D137G+V187A; SPIRPRP+N94K+D96L+L97M+Q249R; SPPPRPRP+N94K+D96L+L97M+Q249R; SPIRPRP-I-D57G+N94K+D96L-I-L97M+Q249R; SPPRRP+I90F+D96L+E99K+V187A; SPIRPRP+D137G+D167G+E21V-I-W221L; E1SPIRPRP+I90F+D96L+E99K-I-V187A; E1SRKRKRK+I90F+D96L+E99K+V187A; E1SPPJKPPJK+I90F+D96L+E99K-I-V187A; E1SPPRRP+D62R+I90F+D96L+E99K+V187A; E1SPPRRP+I90F+D96L+E99K+V187A+N200R+R209A; E1SPPRRP+I90F+D96L+E99K+V187A+T199R+N200R+R209A; ElSP-tRPRP+D57G+D62R+N94K+D96L+Q249R; E1SPIRPRP+D57G-I-N94K+D96L+N200R-I-R209A-1-Q249R; E1SP-CRPRP+D57G+N94K+D96L+T199R+N200R+Q249R; E1SPPRRP+I90F+D96L+E99K+V187A+T199R; ElSPmPRP+D57G+N94K+D96L+T199R+R209A+Q249R; E1SPIRPRP+I90F+D96L+E99K-I-V187A+Q249R; E1SPPRRP+I90F+D96L+E99K+V187A+P253R; E1SPPRRP+I90F+D96L+E99K+D137G+D167G+V187A+Q249R; E1SPPRRP+I90F+D96L+E99K+D137G+V187A+Q249R; E1SPPRRP+D96L+E99K+V187A+Q249R; E1SPPRPR+V2P+N94K+D96L+Q249R; E1SPPWWP+V2W+S3R+N94K+D96L+Q249R; E1SPPWRP+ V2R+S3R+N94K+D96L+Q249R; E1SPPRWP+V2R+S3R+N94K+D96L+Q249R; E1SPPWWP+V2R+S3W+N94K+D96L+Q249R; E1SPPRWP+V2W+S3R+N94K+D96L+Q249R; E1SPPRWP+V2R+S3W+N94K+D96L+Q249R; E1SPPRWP+N94K+D96L+Q249R; E1SPPRRP+N94K+D96L+Q249R; E1APPPRPRPRPRP+V2G+S3T+D57G+N94K+D96L+L97M+Q249R; ElAPPPRTRPRPRS+V2G+S3T+Q4P+D5E+D57G+N94K-l-D96L+L97M+Q249R; or E1APPPKASPRQRP+V2G+D5Q+L6M+D57G+N94K+D96L+L97M+Q249R. SCIRR+N94K+D96L+E239C+Q249R E1SPPRRP+D57G+N94K+D96L+Y53C+K127C+Q249R E1SPPRRPR+V2R+S3P+N94K+D96L+Q249R E1SPPWPRP+V2R+S3P+N94K+D96L+Q249R E1SPPRRP+N94K+D96L+E99K E1SPPRRP+N94K+D96L+E99K+Q249R E1SPPCGRRP+N94K+D96L+E239C+Q249R E1SPCRPRP+N94K+D96L+E239C+Q249R SPPCRRRP+N94K+D96L+E239C +Q249R 35. A method of preparing a mutated lipolytic enzyme, which method comprises at least the following steps: (a) subjecting a DNA sequence encoding a parent Upolytic enzyme to mutagenesis to form a variety of mutated DNA sequences; (b) expressing the mutated DNA sequences in host ceUs; (c) screening for host ceUs expressing a mutated Upolytic enzyme which has a decreased dependence on calcium and/or an improved tolerance towards a detergent or a detergent component as compared to the parent Upolytic enzyme; and (d) selecting a mutated Upolytic enzyme among those resulting from step (c) which, when present in detergent composition A or B, is capable of removing at least 15% more lard from a lard stained swatch, than the same detergent composition without the enzyme, in a one cycle wash assay comprising subjecting 7 lard-stained cotton swatches (9 x 9 cm) per beaker to a one cycle wash in a thermostated TOM, each beaker containing 1000 ml of water comprising 3.2 mM Ca + /Mg 2+ (in a ratio of 5:1) and 5 g/1 of said detergent composition, adjusted to pH 10, and comprising 12500 LU/1 of the Upolytic enzyme, for 20 minutes at a temperature of 30°C, foUowed by rinsing for 15 minutes in running tap water and overnight linedrying at room temperature, subsequent extraction and quantification of fatty matter from the resulting swatches by Soxhlet extraction. 36. The method according to claim 35, wherein the mutagenesis of step (a) is a random mutagenesis, preferably a localized random mutagenesis performed in the lipid contact zone or in any peptide addition present at the N-terminal and/or C-terminal end of the parent lipolytic enzyme. 37. A method according to claim 35 or 36, wherein steps (a), (b), (c) and/or (d) are repeated one or more times. 38. A method of preparing a mutated lipolytic enzyme, which method comprises at least the following steps: (a) constructing mutated DNA sequences by combining i) a DNA sequence encoding a first parent Upolytic enzyme or a part of said DNA sequence and u) a DNA sequence encoding a second parent Upolytic enzyme or a part of said DNA sequence and optionally ui) further DNA sequences encoding a third (and optionally further) parent Upolytic enzymes or a part of said DNA sequence(s), the DNA sequences to be combined being sufficiently homologous to allow for homologous recombination between the sequences to take place, (b) expressing the resulting mutated DNA sequences in host ceUs, and (c) selecting a mutated Upolytic enzyme among those resulting from step (b) which, when present in detergent composition A and/or B is capable of removing at least 15% more lard from a lard stained swatch, than the same detergent composition without the enzyme, in a one cycle wash assay comprising subjecting 7 lard-stained cotton swatches (9 x 9 cm) per beaker to a one cycle wash in a thermostated TOM, each beaker containing 1000 ml of water comprising 3.2 mM Ca 2+ /Mg 2+ (in a ratio of 5:1) and 5 g/1 of said detergent composition, adjusted to pH 10, and comprising 12500 LU/1 of the Upolytic enzyme, for 20 minutes at a temperature of 30°C, foUowed by rinsing for 15 minutes in running tap water and ovemight linedrying at room temperature, subsequent extraction and quantification of fatty matter from the resulting swatches by Soxhlet extraction. 39. The method according to claim 38, wherein prior to step (c) a screening is made for host cells expressing a mutated lipolytic enzyme which has a decreased dependence on calcium and/or an improved tolerance towards a detergent or a detergent component as compared to any of the parent lipolytic enzymes. 40. The method according to claim 38 or 39, wherein one or more of the homologous DNA sequences i)-iii) used for constructing the mutated DNA sequence of step (a) have been prepared by (a) subjecting a DNA sequence encoding a parent Upolytic enzyme to mutagenesis to form a variety of mutated DNA sequences; (b) expressing the mutated DNA sequences in host ceUs; (c) screening for host ceUs expressing a mutated Upolytic enzyme which has a decreased dependence on calcium and or an improved tolerance towards a detergent or a detergent component as compared to the parent Upolytic enzyme; and optionaUy (d) selecting a mutated Upolytic enzyme among those resulting from step (c) which, when present in detergent composition A or B, is capable of removing at least 15% more lard from a lard stained swatch, than the same detergent composition without the enzyme, in a one cycle wash assay comprising subjecting 7 lard-stained cotton swatches (9 x 9 cm) per beaker to a one cycle wash in a thermostated TOM, each beaker containing 1000 ml of water comprising 3.2 mM Ca 2+ /Mg + (in a ratio of 5:1) and 5 g/1 of said detergent composition, adjusted to pH 10, and comprising 12500 LU/1 of the Upolytic enzyme, for 20 minutes at a temperature of 30°C, foUowed by rinsing for 15 minutes in running tap water and overnight Unedrying at room temperature, subsequent extraction and quantification of fatty matter from the resulting swatches by Soxhlet extraction. 41. The method according to any of claims 38-40, wherein the parent lipolytic enzymes encoded " by the homologous DNA sequences i), ii) and iii) constitute different variants of the same naturally occurring lipolytic enzyme. 42. The method according to any of claims 38-41, wherein the parent Upolytic enzyme is derived from a fungus, such as from H. lanuginosa, or from a bacterium, such as a Pseudomonas sp. 43. The method according to any of claims 38-42, wherein the combination of DNA sequences is performed by sexual PCR or gene shuffling. 44. The method according to any of claims 38-43, which comprises an additional step involving culturing a suitable host organism comprising a DNA sequence encoding the first wash mutated lipolytic enzyme selected in step (d) under suitable conditions for expressing the enzyme and recovering the expressed enzyme from the culture medium. 45. A first wash lipolytic enzyme constructed or produced by the method according to any of claims 38-44. 46. A DNA construct comprising a DNA sequence encoding the lipolytic enzyme according to any of claims 1-34 or 45. 47. A DNA comprising a DNA sequence encoding the lipolytic enzyme according to any of claims 1-34 or 45, which DNA sequence is isolated from the host cell screened in the method according to any of claims 38-44. 48. A DNA construct encoding a H. lanuginosa lipolytic enzyme variant according to any of claims 26-34. 49. A vector harbouring a DNA construct according to any of claims 46-48. 50. The vector according to claim 49, which is a plasmid or a bacteriophage. 51. The vector according to claim 49 or 50, which is an expression vector further comprising DNA sequences permitting expression of the lipolytic enzyme. 52. A host ceU harbouring a DNA construct according to any of claims 46-48, or a vector according to any of claims 49-51. 53. The ceU according to claim 52, which is a microbial cell. 54. The ceU according to claim 53, which is a cell of a fungal or a bacterial strain. 55. The cell according to claim 54, which is a cell of the genus Aspergillus, such as A. niger, A. oryzae and A. nidulans, of the genus Fusarium, such as F. graminearum, or a cell of the genus Saccharomyces, e.g. S. cerevisiae. 56. The cell according to claim 54, which is a cell of a gram-positive bacterial strain, e.g. of the * genus Bacillus, such as Bacillus subtilis, Bacillus licheniformis, Bacillus lentus, Bacillus brevis, Bacillus stearothermophilus, Bacillus alkalophilus, Bacillus amyloliquefaciens, Bacillus coagulans, Bacillus circulans, Bacillus lautus, Bacillus thuringiensis or Streptomyces lividans or Streptomyces murinus, or a cell of the genus Pseudomonas, or a cell of a gram-negative bacterial strain, such as E.coli. 51. A method of producing a Upolytic enzyme according to any of claims 1-34, which method comprises preparing a variant Upolytic enzyme in accordance with the method of any of claims 38-44 and recovering the lipolytic enzyme variant from the host ceU screened in step (c). 58. A method of producing a Upolytic enzyme according to any of claims 1-34, which method comprises culturing a host cell according to any of claims 52-56 under suitable conditions to express the enzyme, and recovering the expressed enzyme from the culture. 59. A Upolytic enzyme produced by the method according to claim 57 or 58. 60. A detergent additive comprising a lipolytic enzyme according to any of claims 1-37, optionaUy in the form of a non-dusting granulate, stabilised Uquid or protected enzyme. 61. A detergent additive according to claim 60 which contains 0.02-200 mg of enzyme protein/g of the additive. 62. A detergent additive according to claim 60 or 61, which additionally comprises another enzyme such as a protease, amylase, peroxidase, cutinase, Upase and/or ceUulase. 63. A detergent composition comprising a lipolytic enzyme according to any of claims 1-37. 64. A detergent composition according to claim 63 which additionally comprises another enzyme such as a protease, amylase, peroxidase, cutinase, lipase and/or cellulase. SUBSTITUTE SHEET

Inventors: BORCH KIM, KRETZSCHMAR TITUS, OKKELS JENS SIGURD, PATKAR SHAMKANT ANANT, PETERSEN DORTE AABY, ROYER JOHN C, SVENDSEN ALLAN, THELLERSEN MARIANNE, BORCH, KIM, KRETZSCHMAR, TITUS, OKKELS, JENS, SIGURD, PATKAR, SHAMKANT, ANANT, PETERSEN, DORTE, AABY, ROYER, JOHN, C., SVENDSEN, ALLAN, THELLERSEN, MARIANNE
Applicants: BORCH KIM, KRETZSCHMAR TITUS, NOVONORDISK AS, OKKELS JENS SIGURD, PATKAR SHAMKANT ANANT, PETERSEN DORTE AABY, ROYER JOHN C, SVENDSEN ALLAN, THELLERSEN MARIANNE, BORCH, KIM, KRETZSCHMAR, TITUS, NOVO NORDISK A/S, OKKELS, JENS, SIGURD, PATKAR, SHAMKANT, ANANT, PETERSEN, DORTE, AABY, ROYER, JOHN, C., SVENDSEN, ALLAN, THELLERSEN, MARIANNE
Application year: 1996
Main classification: C12N9
IPC codes: C11D3, C12N9, C12N15