A PROCESS FOR PREPARING PEROXYCARBOXYLIC ACIDS

Abstract: In a process for the preparation of a compound of general formula (I), wherein R is an organic residue, in particular a linear or branched alkyl group, an aryl group or an alkyl aryl group each of which is optionally substituted with one or more hydroxy, alkyloxy, alkoxy, nitro, amino, alkylamino, sulfoxy, sulfono, amido, halo or carboxy groups, a compound of the general formula (II) R-CO-X-R1, wherein X is an oxygen or nitrogen atom, and R1 has the meaning indicated above for R, or is a hydrogen atom or a carbohydrate residue, is reacted with hydrogen peroxide in the presence of a catalytic antibody raised against an analogue of a transition state of the reaction and capable of binding to said transition state. The catalytic antibody may be used as a component of detergents for the generation of peroxycarboxylic acid bleach during washing or as a catalyst for organic synthesis.

Description: A PROCESS FOR PREPARING PEROXYCARBOXYLIC ACIDS FIELD OF INVENTION The present invention relates to a novel process for the preparation of peroxycarboxylic acids. Also, the present 5 invention relates to a catalyst for use in the process, as well as a detergent additive and a detergent composition containing the catalyst. BACKGROUND OF THE INVENTION Peroxycarboxylic acids are frequently used for a number of 10 applications due to their unique oxidizing properties. One such application is the use of peroxycarboxylic acids for bleaching under laundry conditions (e.g. as described in "Chemistry of long-chain peroxy acids under laundry conditions", T.A.B.M. Bols an, R. Kok and A.D. Vreugdenhil, J. Am. Oil Chem. Soc. 15 1988, 65, (7) 1211). In this context, the peroxycarboxylic acid is usually formed in situ, by the reaction of a precursor (an "activator", e.g. tetraacetyl-ethylenediamine (TAED) or nonanoyloxybenzenesulphonate (NOBS) , as described in the reference cited above) with a perhydroxyl anion. 20 Peroxycarboxylic acids are also commonly employed in the field of organic synthesis, as a wide variety of organic molecules may be oxidized by means of these reagents. The various uses of peroxycarboxylic acids are exhaustively described in the art, e.g. in Comprehensive Organic Chemistry edited by Barton and 25 Ollis, Pergamon Press, 1979. Some peroxycarboxylic acids such as meta-chloro-perbenzoic acid have become commercially available as a result of their wide applicability. Even though some peroxycarboxylic acids are useful and commercially available reagents their use is limited because of 0 their relatively high cost and the risks (in particular the explosion hazard) involved in handling the reagents, especially when this is done on a large scale. Accordingly, there is a need for improved methods of preparing peroxycarboxylic acids as well as for methods which make it safer to use them in organic synthesis. DISCLOSURE OF THE INVENTION The present invention relates to a process for the preparation of a compound of the general formula (I) 0 II R-C (I) I O-OH wherein R is an organic residue, in particular a linear or branched alkyl group, an aryl group or an alkyl aryl group each of which is optionally substituted with one or more hydroxy, alkoxy, nitro, amino, alkylamino, sulfonato, sulfo, a ido, halo or carboxy groups, the process comprising reacting a compound of the general formula (II) R-CO-X-R, (II) wherein X is an oxygen or nitrogen atom, and R 1 has the meaning indicated above for R, or is a hydrogen atom or a carbohydrate residue, with hydrogen peroxide in the presence of a catalytic antibody raised against an analogue of a transition state of the reaction and capable of binding to said transition state. In the present context, the term "catalytic antibody" is intended to indicate an antibody which has the catalytic activity of an enzyme in that it is able to bind selectively to the transition state of a given reaction whereby the transition state is stabilized. In this way, the activation energy required for the reaction in question is decreased, and the rate of the reaction is consequently increased. A transition state may be defined as an unstable intermediate which cannot be isolated as it is extremely short-lived. Consequently, it is not possible to raise antibodies against it. An analogue of a transition state may be defined as a stable substance which has those characteristics of the corresponding transition state, such as charge or configuration, which provide it with identical or nearly identical antigenic determinant(s) so that catalytic antibodies raised against the analogue are also able to bind to the transition state itself when present in a reaction so as to provide a catalytic effect. The preparation and use of catalytic antibodies is described in, e.g., A. Tramontano, A.A. A mann and R.A. Lerner, J. Am. Chem. Soc. 110, 1988, pp. 2282-2286; A. Tramontano, K.D. Janda and R.A. Lerner, Science 234, 19 Dec. 1986, pp. 1566-1570; S.J. Pollack, J.W. Jacobs and P.G. Schultz, Science 234 , 19 Dec. 1986, pp. 1570- 1573; A. Tramontano, K.D. Janda and R.A. Lerner, Proc. Natl. Acad. Sci. USA 83, Sep. 1986, pp. 6736-6740; US 4,659,567; US 4,792,446; EP 305 870; EP 260 939 and WO 88/09380. None of these publications disclose the use of catalytic antibodies for the preparation of peroxycarboxylic acids, and this use is therefore believed to be novel. The process of the invention may be summarized as indicated in Reaction Scheme (A) : Antibody O l\ R-CO-X-R. (II) R-C (I) (A) / OOH /~*1 H 2 0 2 H 2 0 ^— H ** ,202, precursor In Reaction Scheme (A) , R, R 1 and X are as defined above. In particular, R may be a linear alkyl group, in particular a linear alkyl group with 1-19 carbon atoms, preferably 2-13 carbon atoms, more preferably 5-11 carbon atoms, and most preferably 7-9 carbon atoms. If the compound (II) is an optically active compound, it is possible to employ a racemate or one of the enantio er forms of the compound (II) for reaction with hydrogen peroxide. The process of the invention may thus be employed for the synthesis of optically active peroxycarboxylic acids. In one embodiment of the process of the invention, X may be oxygen, and R 1 may be a carbohydrate residue, preferably a monosaccharide, more preferably a methyl, ethyl or propyl glycoside, and most preferably a methyl, ethyl or propyl glucoside. Alternatively, R., may be an aryl group which is preferably substituted with one or more sulfoxy, sulfono or hydroxy groups. In the reaction of compound (II) with hydrogen peroxide, the transition state may, for instance, be the following tetrahedral structure 0 " I R-C-OR, I 1 00H wherein R and R 1 are as indicated above. In this case, suitable transition state analogs useful for raising appropriate antibodies are phosphonate esters of the general formula (III) / R-P-OR (III) ft 0 wherein R and R 1 are as indicated above. These analogs differ from the transition state itself by being stable compounds which may be employed to raise antibodies capable of binding the transition state of the reaction described above. Examples of specific phosphonate esters for use in the present process arep-nitrophenyl-(4-carboxybutane) phosphonate, p-sulfophenyl- l(4-carboxybutane) phosphonate or 1'-0-ethylglucosyl-(4- carboxybutane) phosphonate. It should be noted, however, that phosphonate esters are relatively small molecules and may not in themselves be used for immunization purposes. In order to have an immunizing effect, they should therefore be bound (either directly or by means of a linking group such as a mercaptan, a succinimide or the like) to a suitable carrier molecule such as bovine serum albumin or keyhole limpet he ocyanin to form a conjugate which may then be used for immunization. Transition state analogue- carrier conjugates may, for instance, be produced by the method described in WO 88/09380. The antibody employed in the process of the invention may be a polyclonal antibody. Polyclonal antibodies may be prepared by immunizing a suitable animal (such as rabbits, goats, horses, sheep, mice, chickens, rats and guinea pigs) with the transition state analogue (in particular phoshonate ester) carrier conjugate (suitably dissolved or dispersed in a physiologically acceptable solvent or diluent) and isolating the antibody from the serum in a manner known per se (cf. A. Johnstone and R. Thorpe, I munochemistry in Practice, 2nd Ed. Blackwell Scientific publications, 1987, pp. 30-34 and 48-55). It is, however, usually preferred to employ monoclonal antibodies as these have become easy to produce in large quantities and in a high degree of purity. Furthermore, they exhibit great uniformity. Alternatively, it is possible to use a fragment of a monoclonal antibody, e.g. a Fab 1 , F(ab') 2 or other fragment. Monoclonal antibodies may be prepared by fusing cells producing the antibody with myeloma cells of an established cell line, selecting and cloning the resulting hybridoma cells and growing them in a suitable medium to produce the antibody and isolating the antibody from the culture, e.g. as described in A. Johnstone and R. Thorpe, op. cit. pp. 35-43. The antibody may also be produced by the method described in WO 88/09380. A further method of producing antibodies may be to cultivate a host cell transformed with a recombinant DNA vector which carries a DNA sequence encoding the antibody or a fragment thereof as well as DNA sequences encoding functions permitting 5 the expression of the DNA sequence encoding the antibody, in a culture medium under conditions permitting the expression of the antibody and recovering the antibody from the culture. Preparation of antibodies by recombinant DNA techniques is, for instance, described in M. Better et al., Science 240, 1988, p. 101041; and CR. Wood et al. , Nature 314, 1985, p. 446. A DNA fragment encoding the antibody may, for instance, be isolated by establishing a cDNA or geno ic library of an antibody-producing cell line and screening for positive clones by conventional procedures such as by hybridization to 15 oligonucleotide probes synthesized on the basis of the full or partial amino acid sequence of the antibody or by selecting for clones producing an antibody which is reactive with the appropriate transition state analog. Once selected, the DNA sequence may be inserted into a suitable 0 replicable expression vector comprising appropriate promotor, operator and terminator sequences permitting the antibody to be expressed in a particular host organism, as well as an origin of replication enabling the vector to replicate in the host organism in question. 5 The resulting expression vector may then be transformed into a suitable host cell, such as a fungal cell, preferred examples of which are a species of Asperαillus, most preferably Asperqillus oryzae or Aspercrillus niger. Fungal cells may be transformed by a process involving protoplast formation and 0 transformation of the protoplasts followed by regeneration of the cell wall in a manner known per se. The use of Asperqillus as a host microorganism is described in EP 238,023 (of Novo In- dustri A/S). The host cell may also be a yeast cell, such as a strain of Saccharo yces spp. or Schizosaccharo yces spp. , in particular Saccharomyces cerevisiae. Alternatively, the host organism may be a bacterium, in particular strains of Streptomyces and Bacillus, or E. coli. The transformation of bacterial cells may be performed according to conventional methods, e.g. as described in Sambrook, Fritsch and Maniatis, Molecular Cloning: A Labora¬ tory Manual, Cold Spring Harbor, New York 1989. The screening of appropriate DNA sequences and construction of vectors may also be carried out by standard procedures, cf. Sambrook, Fritsch and Maniatis et al., op. cit. The medium used to cultivate the transformed host cells may be any conventional medium suitable for growing the host cells in question. The expressed antibody may conveniently be secreted into the culture medium and may be recovered therefor by well- known procedures including separating the cells from the medium by centrifugation or filtration, precipitating proteinaceous components of the medium by means of a salt such as ammonium sulphate, followed by chromatographic procedures such as ion exchange chromatography, affinity chromatography, or the like. In another aspect, the present invention relates to a catalyst for use in the process according to the invention, which catalyst comprises a catalytic antibody raised against an analogue of a transition state of the reaction of a compound of the general formula II R-CO-X-R. (II) wherein R and R 1 are as indicated above, with hydrogen peroxide, said antibody being capable of binding said transition state. Other characteristics of the catalyst appear from the description of the catalytic antibody above. In a further aspect, the present invention relates to a detergent additive which comprises the catalyst of the invention. The detergent additive of the invention may suitably be in the form of a non-dusting granulate, a liquid, in particular a stabilized liquid, or a protected antibody. Non- dusting granulates may be produced in a manner analogous to that of producing enzyme granulates, e.g., as disclosed in US 4,106,991 and 4,661,452 (both to Novo Industri A/S) and may optionally be coated by methods known in the art. Liquid antibody preparations may, for instance, be stabilized by adding a polyol such as propylene glycol, a sugar or sugar alcohol, lactic acid or boric acid according to established methods. Other stabilizers are well known in the art. Protected antibodies may be prepared according to the method disclosed in EP 238,216 (for enzymes). The detergent additive may further include one or more enzymes, such as proteases, lipases, cellulases, peroxidases or amylases or a combination thereof, conventionally included in detergent additives. In this case, the catalytic antibody included in the detergent additive should be one which is resistant to the action of such enzymes, in particular that of protease. This may be accomplished by selection for antibodies which are stable in the presence of protease, by formulating the antibody so that the contact with protease will be minimized (e.g. as a granulate) , or by modifying the protease cleavage site(s) on the antibody, e.g. by protein engineering in a manner known per se. In a still further aspect, the present invention relates to a detergent composition which comprises a catalyst according to the invention. The catalyst may preferably be present in an amount corresponding to 0.01-100 mg of catalyst per litre of washing liquors. In one advantageous embodiment, the detergent composition additionally comprises a compound of the general formula II R-CO-X-R, (II) wherein R, R 1 and X are as indicated above. In this way, it is possible to generate peroxycarboxylic acid bleaching agents in situ during washing rather than including them during manufacture of the detergent composition. The detergent composition of the invention may be formulated in any convenient form, e.g. as a powder or liquid. The antibody may be stabilized in a liquid detergent by the inclusion of stabilizers as indicated above. The detergent composition may further include hydrogen peroxide precursors such as perborates or percarbonates. Usually, the pH of a solution of the detergent composition of the invention will be in the range of 7-12 and in some instances in the range of 7.0-10.5. Detergent enzymes such as proteases, lipases, cellulases or amylases or a combination of two or more of these may be included in the detergent compositions of the invention, either separately or in a combined additive as described above. In this case, the antibody should be one which is resistant to the action of such enzymes, notably that of protease as explained above. The detergent composition of the invention may comprise one or more surface-active agents, such as anionic surfactants (e.g. linear alkyl benzene sulfonates, fatty alcohol sulfates, fatty alcohol ether sulfates, α-olefin sulfonates or soaps) , non¬ ionic surfactants (e.g. fatty alcohol ethoxylates, nonylphenol ethoxylates, fatty acid esters of sucrose and glucose, alkyl glycosides or esters of polyoxyethoxylated alkyl glycosides) , cationic surfactants and/or zwitterionic surfactants. Liquid and powder detergents may be formulated substantially as described in "Frame formulations for liquid/powder heavy-duty detergents" in J. Falbe, Surfactants in Consumer Products. Theory, Technology and Application, Springer Verlag, 1987. Thus, as described in Falbe, op. cit. , a liquid heavy-duty detergent may comprise anionic surfactants, non-ionic surfactants, suds controlling agents, enzymes, foam boosters, builders, formulation aids, optical brighteners, stabilizers, fabric softeners, fragrances, dyestuffs and water. Similarly, a powder heavy-duty detergent may comprise anionic surfactants, non-ionic surfactants, suds controlling agents, foam boosters, chelating agents, ion exchangers, alkalis cobuilders, bleaching agents, bleach activators, bleach stabilizers, fabric softeners, antiredeposition agents, enzymes, optical brighteners, anticorrosion agents, fragrances, dyestuffs and blueing agents, formulation aids, fillers and water. For organic synthesis according to the invention, the catalytic antibody may be used per se. It is, however, possible to immobilize the antibody in order to facilitate the recovery of the peroxycarboxylic acids produced by the present process. The immobilization procedures used to immobilize the antibody may be analogous to the established procedures for immobilizing enzymes (cf. for instance K. Mosbach, ed. , "Immobilized Enzymes", Methods in Enzy ology 44, Academic Press, New York, 1976) and include cross-linking of cell homogenates, covalent coupling to insoluble organic or inorganic supports, entrapment in gels and adsorption to ion exchange resins or other adsorbent materials. Coating on a particulate support may also be employed (cf. for instance A.R. Macrae and R.C. Hammond, Biotechnology and Genetic Engineering Reviews 3, 1985, p. 193) . Suitable support materials for the immobilized antibody are, for instance, plastics (e.g. polypropylene, polystyrene, polyvinylchloride, polyurethane, latex, nylon, teflon, dacron, polyvinylacetate, polyvinylalcohol or any suitable copolymer thereof) , polysaccharides (e.g. agarose or dextran) , ion exchange resins (both cation and anion exchange resins) , silicon polymers (e.g. siloxane) or silicates (e.g. glass) . It is preferred to immobilize the antibody on an ion exchange resin by adsorbing the antibody to the resin or by cross- linking it to the resin by means of glutaraldehyde or another cross-linking agent in a manner known per se. A particularly preferred resin is a weakly basic anion exchange resin which may be a polystyrene-, acrylic- or phenyl-formaldehyde-type resin. Examples of commercially available polyacrylic-type resins are Lewatit E 1999/85 (registered trademark of Bayer, Federal Republic of Germany) and Duolite ES-568 (registered trademark of Rohm & Haas, FRG) . Immobilization of antibodies to this type of resin may be carried out according to EP 140 542 (disclosing the immobilization of enzymes) . Immobilization to phenyl-formaldehyde-type resins may be done according to the procedure of DK 85/878 (disclosing the immobilization of enzymes) . An example of a commercially available acrylic-type resin is Lewatit E2001/85 (registered trademark of Bayer, FRG) . Another convenient material for immobilizing antibodies is an inorganic support, such as a silicate. The antibody may be attached to the support by adsorption cr by covaient coupling, eg. as described in K. Mosbach, ed. , op.cit. (for enzymes). The process according to the method of the invention may be carried out in the compound (II) itself (which in this case also acts as a solvent) or in solvents such as water, aqueous buffer solutions or organic solvents. Some preferred organic solvents are hydrocarbons such as hexane, cyclohexane, heptane, benzene and toluene, methylene chloride and hexachloroethane, acetonitrile, dimethylforma ide, dioxane and tetrahydrofurane. It is preferred to employ solvents in which the compound (II) and products of the reaction are highly soluble and in which the antibody maintains a good stability and activity. The temperature at which the reaction of the compound (II) takes place is not believed to be critical, but may conveniently be in the range of about 20-100 °C, such as about 30-80 °C. The hydrogen peroxide employed according to the process of the invention may be added as such to the reaction mixture either at the beginning of the reaction or at a desired rate in the course of the reaction. Alternatively, it is possible to provide a precursor of hydrogen peroxide, such precursors being compounds which give rise to the generation of hydrogen peroxide in situ for reaction with the compound (II) , such as percarbonates or perborates. The water generated in the course of the process according to the invention (i.e. when R 1 is hydrogen) may, if desired, be removed by methods known in the art such as by distillation, exposure to dessicants etc. The invention is further illustrated by the following example which is not in any way intended to limit the scope of the invention as claimed. EXAMPLE. General methods Peroxycarboxylic acid references were prepared according to the method described by W.E. Parker, C. Ricciuti, CL. Ogg and D. Swern, J. Am. Chem. Soc. 1955, 77, 4037. Percarboxylic acid levels may be monitored, also when hydrogen peroxide is present together with the peroxycarboxylic acid, by methods known in the art, e.g. by iodometry at 5 * C, as described by Sully and Williams in Analyst, 1962, 87, 653. Hydrolysis of p-nitrophenol butyrate was monitored spectrophoto etrically by monitoring the absorbance at 400 nm using a Hewlett Packard HP 8452A diode array spectrophotometer. Preparation and testing of catalytic antibodies capable of catalyzing the perhydrolysis of p-nitrophenol butyrate by hydrogen peroxide (as shown in Reaction Scheme I below) : Synthesis of transition state analogs: Preparation of diethyl 4- (ethyloxycarbonyl) butanephosphonate (1) PiOEt: ethyl-5-bromo pentanoate (30g, 0.14mol) and triethylphosphite (50g, 0.3mol) was heated at 160 ° C overnight. The crude mixture was then distilled, yielding 30g of the title compound. 1 H NMR, CDC1 3 ,<5= 4.2-4.0 (m,6H) , 2.32 (t,2H) , 1.8-1.55 (m, 6H) , 1.32 (t,6H) , 1.25 (t,3H) . Preparation of ethyl 4 - (ethyloxycarbonyl ) butane chlorophosphonate (2 ) : To diethyl 4-(ethyloxycarbonyl) butanephosphonate (lOg, 38mmol) in CC1 4 (20ml) was added PC1 5 (7.9g, 38mmol) . The mixture was refluxed overnight after which the solvent was removed. The crude title compound thus obtained was used as such in the next step.. Preparation of -ethyl p-nitrophenyl 4-carboethoxybutyl phosphonate (3) : Sodium (0.79g, 34.5mmol) was added to ethanol (30 ml, 99.9%). When the reaction was complete, toluene (120ml) and then p- nitrophenol (4.4g, 31.4mmol) were added. The ethanol was distilled off, the reaction cooled to 0 °C and a toluene (30ml) solution of compound 2 (7.5g 31.4mmol) was added slowly. After 1.5 h at 75 °C the precipitate was filtered off, and the solution was extracted with NaOH (1.0M) . The product was further purified by chromatography, yielding 2.3 g of product. 1 H NMR, CDC1 3 ,<5= 8.22 (d,2H), 7.37 (d,2H), 4.3-4.05 (m,4H), 2.34 (t,2H) , 2.0-1.9 (m,2H), 1.32-1.15 (m,4H) , 1.3 (t,3H) , 1.25 (t,3H) . Preparation of p-nitrophenyl- (4-carboxybutan) phosphonate (4): KOH (3g) in ethanol (60ml) was added to compound 3 (2.3g) and heated to 120 e C for 2 h. The mixture was acidified with HC1 (6 N) , extracted with diethyl ether and dried over MgS0 4 . The product was purified by repeated precipitations from diethyl ether with n-pentane, yielding 1 g of product. 1 H NMR, D 2 0,<5=8.32 (d,2H) , 7.36 (d,2H) , 2.47 (t,2H), 1.85 (m,2H) , 1.7 (m,4H) . Preparation of i munogenic conjugate: The selected carrier was KLH (Keyhole Limpet Hemocyanin) . The conjugate was prepared by adding N-hydroxysuccinimide (4mg) , dicyclohexyl carbodiimide (7.4mg) and pyrrolidinopyridin (0.4mg) in CH 2 C1 2 (2ml) to compound 4 (lOmg) . The mixture was allowed to react for 20 h at 20°C The dicyclohexyl urea formed was removed by filtration and KLH (5.3mg) in H 2 0 (1ml) was added slowly. The reaction proceeded for 2 h. The product was dialyzed 24 h against H 2 0. The resulting product had 0.29 mg hapten coupled to 5,3 mg KLH (monitored by hydrolysis of p- nitrophenol) . Immunization of mice: Four RBF mice were immunized three times at bi-weekly intervals. In the first immunization each mouse was injected subcutaneously with 100 μl of conjugate mixed with 100 μl of Freunds's complete adjuvant (corresponding to 0.4μg of compound 4) . The two following immunizations had 100 μl of conjugate mixed with 100 μl of Freunds • s incomplete adjuvant (corresponding to 2 μg of compound 4) . Eyeblood samples were redrawn from all four mice and serum was analyzed by ELISA using hapten coupled to BSA as antigen (Hapten was coupled to BSA following the same principles as with KLH) . All four mice showed good titers. Eight weeks after the last immunization one mouse received an intravenous booster injection (corresponding to 2 μg of compound 4) and three days after that the spleen was removed. Cell fusion and culture of cells: Monoclonal antibodies were prepared by standard techniques first described by Kohler and Milstein, Nature 1975, 256, 495. Thus, the spleen from one mouse was removed and carefully dissected and disrupted. Fusion with sp2 myeloma cells was carried out in the presence of polyethylene glycol. The resulting hybridoma cells were seeded in 960 microtiter wells using selective medium (HAT) . After two weeks of growth, the supernatants were analyzed in the above mentioned ELISA. Fourty-eight selected hybridomas were further screened in an ELISA where BSA-coupled and free hapten were allowed to compete for the antibodies. Among these, seven hybridomas were selected, and monoclonal antibodies were obtained from the hybridoma tissue cultures. Purification of monoclonal antibodies: Hybridoma supernatants were concentrated, the pH was adjusted to 8.5 and the monoclonal antibodies were then purified by adsorption on to Protein G Sepharose and eluted with citrate buffer pH 3.0. The monoclonal antibodies were all of IgG 2 subclass. Evaluation of catalysis: Catalysis was monitored in an assay using 1 ml phosphate buffer (50 mM, pH 8 or 9) containing 0.5 mM p-nitrophenyl butyrate and 1.5 mM H 2 0 2 and 2 μM antibody (if present) . The reactions were initiated by addition of p-nitrophenol butyrate. Table 1 below shows the catalysis obtained with three different monoclonal antibodies, namely F37A12, F35B9 and F48A6. Of these, F37A12 and F48A6 catalyses both the hydrolysis and the perhydrolysis of p-nitrophenyl butyrate. F35B9 does not catalyze the reaction at all, presumably because it preferentially binds the starting material. Table 1. Catalysis measured as formation of peroxybutanoic acid and release of p-nitrophenol. Antibody pH Time Total hydro¬ Perhydrolysis, lysis measured measured as: as: [p-nitro¬ [Peroxybutanoic phenol] acid] Blank 80 s 35±2 μM 26±8 μM n=5 F37A12 80 s 88±2 μM 45±5 μM n=2 F35B9 80 s 33±2 μM 23±5 μM n=2 F48A6 !0 s 89±2 μM 45±5 μM n=2 Blank 25 Ξ 32±2 μM 36±7 μM n=5 F37A12 25 s 71±2 μM 61+5 μM n=2 F35B9 25 s 25*2 μM 26±5 uM n=2 F48A6 25 s 60±2 μM 59±5 UM n=2 These examples clearly demonstrate that catalytic antibodies, prepared according to the present invention, are capable of catalyzing perhydrolysis reactions. Thus, at pH 9, the antibody F48A6 is capable of catalyzing the formation of 98% peroxybutanoic acid relative to the total amount of p- nitrophenol released. The superior performance of this antibody compared to the other two antibodies tested, clearly demonstrates that antibodies not disfavoring perhydrolysis over normal hydrolysis can indeed be prepared by the procedure claimed.

Claims: CLAIMS 1. A process for the preparation of a compound of the general formula (I) 0 R-C (I) I O-OH wherein R is an organic residue, in particular a linear or branched alkyl group, an aryl group or an alkyl aryl group each of which is optionally substituted with one or more hydroxy, 10 alkoxy, nitro, amino, alkylamino, sulfonato, sulfo, amido, halo or carboxy groups, the process comprising reacting a compound of the general formula (II) R-CO-X-R, (II) wherein X is an oxygen or nitrogen atom, and R. has the meaning 15 indicated above for R, or is a hydrogen atom or a carbohydrate residue, with hydrogen peroxide in the presence of a catalytic antibody raised against an analogue of a transition state of the reaction and capable of binding to said transition state. 202. A process according to claim 1, wherein R is a linear alkyl group, in particular a linear alkyl group with 1-19 carbon atoms, preferably 2-13 carbon atoms, more preferably 5-11 carbon atoms, and most preferably 7-9 carbon atoms. 3. A process according to claim 1, wherein X is oxygen, and R 1 25 is a carbohydrate residue, preferably a monosaccharide residue, more preferably a methyl, ethyl or propyl glycoside, and most preferably a methyl, ethyl or propyl glucoside. 4. A process according to claim 1, wherein R 1 is an aryl group which is preferably substituted with one or more sulfoxy, sulfono or hydroxy groups. 5. A process according to claim 1, wherein the catalytic 5 antibody is one raised against a phosphonate ester of the general formula (III) 0 I R-P-OR (III) a 0 10 wherein R and R 1 are as indicated in claim 1. 6. A process according to claim 1 or 5, wherein the antibody is a polyclonal antibody, a monoclonal antibody or a fragment thereof. 7. A process according to claim 6, wherein the antibody is an 15 immobilized antibody. 8. A process according to -claim 1, wherein a precursor of hydrogen peroxide, e.g. a perborate or a percarbonate, is provided to generate hydrogen peroxide for the reaction with the compound (II) . 209. A catalyst for use in the process according to claim 1, which comprises a catalytic antibody raised against an analogue of a transition state of the reaction of a compound of the general formula II R-CO-X-R T (II) 25 wherein R and R, are as indicated in claim 1, with hydrogen peroxide, said antibody being capable of binding to said transition state. 10. A catalyst according to claim 9, wherein the catalytic antibody is one raised against a phosphonate ester of the general formula (III) 0 I R-P-OR (III) II 0 wherein R, R 1 and X are as indicated in claim 1. 11. A catalyst according to claim 9 or 10, wherein the antibody is a polyclonal antibody, a monoclonal antibody or a fragment 10 thereof. 12. A catalyst according to claim 11, wherein the antibody is an immobilized antibody. 13. A detergent additive which comprises a catalyst according to any of claims 9-12. 15 14. A detergent composition which comprises a catalyst according to any of claims 9-12. 15. A detergent composition according to claim 14, which comprises the catalyst in an amount corresponding to 0.01-100 mg of catalyst per litre of washing liquors. 20 16. A detergent composition according to claim 14, which additionally comprises a compound of the general formula II R-CO-X-R. (II) wherein R, R 1 and X are as indicated in claim 1. 17. A detergent composition according to claim 15 which further 25 comprises a precursor of hydrogen peroxide, such as a perborate or percarbonate. 18. A detergent composition according to any of claims 14-16 which further comprises an enzyme, e.g. an enzyme selected from the group consisting of proteases, lipases, amylases and cellulases, or a combination of two or more of these enzymes.

Inventors: BJOERKLING FREDERIK, CHRISTENSEN LISELOTTE BJERRE, GODTFREDSEN SVEN ERIK, KIRK OLE, BJOERKLING, FREDERIK, CHRISTENSEN, LISELOTTE, BJERRE, GODTFREDSEN, SVEN, ERIK, KIRK, OLE
Applicants: NOVONORDISK AS, NOVO NORDISK A/S
Application year: 1990
Main classification: C07K15
IPC codes: B01J31, C07B61, C07C407, C07C409, C07K16, C11D3, C11D7, C12N5, C12N9, C12N15, C12P7, C12P21, C12R1