Patent Publication Number: US-2019185791-A1

Title: Use of enzymes, cleaning composition and method for washing

Description:
FIELD OF THE INVENTION 
     The present invention concerns the use of enzymes for preventing, reducing or removing odor, a cleaning composition comprising enzymes and a method for washing an item. 
     REFERENCE TO A SEQUENCE LISTING 
     This application contains a Sequence Listing in computer readable form. The computer readable form is incorporated herein by reference. 
     BACKGROUND OF THE INVENTION 
     Use of enzymes in dishwashing detergents is well known in the field of both automatic dishwashing (ADW) formulas, and in hand dishwashing formulas. Typically proteases and amylases are used in commercial dishwashing detergents. These enzymes are useful for degrading protein and starch/amylose, respectively. However, other kinds of food material than protein and amylose left on dishes may is not degraded by protease and amylase and will remain in the filter of the dishwashing machine. This can give rise to development of undesirable odorous compounds. 
     SUMMARY OF THE INVENTION 
     The present invention concerns the use of at least one enzyme for preventing, reducing or removing odor from an item, wherein the at least one enzyme is selected from an alkaline cellulase and/or an enzyme having mannanase activity. 
     The invention further concerns a cleaning composition comprising a builder and at least one enzyme for preventing, reducing or removing odor from an item, wherein the at least one enzyme is selected from an alkaline cellulase and/or an enzyme having mannanase activity. 
     Further is invented a method for washing an item, wherein the method comprises the steps of:
         (i) Exposing the item to a wash liquor comprising at least one enzyme selected from an alkaline cellulase and/or an enzyme having mannanase activity or the dishwashing composition according to the invention; and   (ii) Rinsing the item with water optionally comprising a rinsing aid;
 
wherein the item is a dishware or a hard surface.
       

     Definitions 
     Carbohydrate oxidase: EC 1.1.3.10 systematic name: pyranose:oxygen 2-oxidoreductase, A flavoprotein (FAD). Also oxidizes D-xylose, L-sorbose and D-glucono-1,5-lactone, which have the same ring conformation and configuration at C-2, C-3 and C-4. 
     Cellobiose oxidase: EC 1.1.3.25, EC 1.1.99.18 Generally this can be stated as: cellobiose+acceptor F⇄cellobiono-1,5-lactone+reduced acceptor, if dioxygen is the acceptor the product is hydrogen peroxide. 
     Detergent components: The term “detergent components” is defined herein to mean the types of chemicals which can be used in detergent compositions for automatic dishwashing. Examples of detergent components are polymers, bleaching systems, bleach activators, bleach catalysts, silicates, dyestuff and metal care agents. 
     Dishware: The term dish ware is intended to mean any form of kitchen utensil, dinner set or tableware such as but not limited to pans, plates, cops, knives, forks, spoons, porcelain etc. 
     Dish wash: The term “dish wash” refers to all forms of washing dishes, e.g. by hand (MDW) or automatic dish wash (ADW). Washing dishes includes, but is not limited to, the cleaning of all forms of crockery such as plates, cups, glasses, bowls, all forms of cutlery such as spoons, knives, forks and serving utensils as well as ceramics, plastics, metals, china, glass and acrylics. 
     Dish washing composition: The term “dish washing composition” refers to compositions intended for cleaning dishware such as plates, cups, glasses, bowls, cutlery such as spoons, knives, forks, serving utensils, ceramics, plastics, metals, china, glass and acrylics in a dishwashing machine. The terms encompass any materials/compounds selected for household or industrial washing applications and the form of the product can be liquid, powder or granulate. In addition enzymes, the automatic dishwashing composition contains detergent components such as polymers, bleaching systems, bleach activators, bleach catalysts, silicates, dyestuff and metal care agents. The dishwashing composition can be use in manual dishwashing (MDW) or automatic dishwashing (ADW). 
     Endoglucanase: The term “endoglucanase” means an endo-1,4-(1,3;1,4)-beta-D-glucan 4-glucanohydrolase (E.C. 3.2.1.4) that catalyzes endohydrolysis of 1,4-beta-D-glycosidic linkages in cellulose, cellulose derivatives (such as carboxymethyl cellulose and hydroxyethyl cellulose), lichenin, beta-1,4 bonds in mixed beta-1,3 glucans such as cereal beta-D-glucans or xyloglucans, and other plant material containing cellulosic components. Endoglucanase activity can be determined by measuring reduction in substrate viscosity or increase in reducing ends determined by a reducing sugar assay (Zhang et al., 2006,  Biotechnology Advances  24: 452-481). For purposes of the present invention, endoglucanase activity is determined using carboxymethyl cellulose (CMC) as substrate according to the procedure of Ghose, 1987,  Pure and Appl. Chem.  59: 257-268, at pH 5, 40° C. 
     Glucose oxidase: The glucose oxidase enzyme (GOx) also known as notatin (EC number 1.1.3.4) is an oxido-reductase that catalyses the oxidation of glucose to hydrogen peroxide and D-glucono-δ-lactone. 
     Hard surface cleaning: The term “Hard surface cleaning” is defined herein as cleaning of hard surfaces wherein hard surfaces may include floors, tables, walls, roofs etc. as well as surfaces of hard objects such as cars (car wash) and dishes (dish wash). Dish washing includes but are not limited to cleaning of plates, cups, glasses, bowls, cutlery such as spoons, knives, forks, serving utensils, ceramics, plastics, metals, china, glass and acrylics. 
     Sequence identity: The relatedness between two amino acid sequences or between two nucleotide sequences is described by the parameter “sequence identity”. 
     For purposes of the present invention, the sequence identity between two amino acid sequences is determined using the Needleman-Wunsch algorithm (Needleman and Wunsch, 1970,  J. Mol. Biol.  48: 443-453) as implemented in the Needle program of the EMBOSS package (EMBOSS: The European Molecular Biology Open Software Suite, Rice et al., 2000,  Trends Genet.  16: 276-277), preferably version 5.0.0 or later. The parameters used are gap open penalty of 10, gap extension penalty of 0.5, and the EBLOSUM62 (EMBOSS version of BLOSUM62) substitution matrix. The output of Needle labeled “longest identity” (obtained using the—nobrief option) is used as the percent identity and is calculated as follows: 
       (Identical Residues×100)/(Length of Alignment−Total Number of Gaps in Alignment)
 
     For purposes of the present invention, the sequence identity between two deoxyribonucleotide sequences is determined using the Needleman-Wunsch algorithm (Needleman and Wunsch, 1970, supra) as implemented in the Needle program of the EMBOSS package (EMBOSS: The European Molecular Biology Open Software Suite, Rice et al., 2000, supra), preferably version 5.0.0 or later. The parameters used are gap open penalty of 10, gap extension penalty of 0.5, and the EDNAFULL (EMBOSS version of NCBI NUC4.4) substitution matrix. The output of Needle labeled “longest identity” (obtained using the—nobrief option) is used as the percent identity and is calculated as follows: 
       (Identical Deoxyribonucleotides×100)/(Length of Alignment−Total Number of Gaps in Alignment)
 
     Variant: The term “variant” means a polypeptide having enzyme activity comprising an alteration, i.e., a substitution, insertion, and/or deletion, at one or more (e.g., several) positions. A substitution means replacement of the amino acid occupying a position with a different amino acid; a deletion means removal of the amino acid occupying a position; and an insertion means adding an amino acid adjacent to and immediately following the amino acid occupying a position. 
     Wash cycle: The term “wash cycle” is defined herein as a washing operation wherein dishware are exposed to the wash liquor for a period of time by circulating the wash liquor and spraying the wash liquor onto the dishware in order to clean the dishware and finally the superfluous wash liquor is removed. A wash cycle may be repeated one, two, three, four, five or even six times at the same or at different temperatures. Hereafter the dishware is generally rinsed and dried. One of the wash cycles can be a soaking step, where the dishware is left soaking in the wash liquor for a period. 
     Wash liquor: The term “wash liquor” is intended to mean the solution or mixture of water and detergents optionally including enzymes used for dishwashing. 
     Overview of Sequences Listing 
     SEQ ID NO: 1 is the amino acid sequence of a cellulase. SEQ ID NO: 2 is the amino acid sequence of a mannanase.
 
SEQ ID NO: 3 is the amino acid sequence of a glucose oxidase.
 
SEQ ID NO: 4 is the amino acid sequence of a cellobiose oxidase.
 
SEQ ID NO: 5 is the amino acid sequence of an amylase.
 
SEQ ID NO: 6 is the amino acid sequence of a protease.
 
SEQ ID NO: 7 is the amino acid sequence of a protease.
 
    
    
     DETAILED DESCRIPTION OF THE INVENTION 
     The invention concerns the use of at least one enzyme for preventing, reducing or removing odor from an item, wherein the at least one enzyme is selected from an alkaline cellulase and/or an enzyme having mannanase activity. 
     The invention further concerns a dishwashing composition comprising a builder and at least one enzyme for preventing, reducing or removing odor from an item, wherein the at least one enzyme is selected from an alkaline cellulase and/or an enzyme having mannanase activity. 
     In accordance with the invention the enzymes or the dishwashing composition can be used in a method for washing an item, wherein the method comprises the steps of:
         a) Exposing the item to a wash liquor comprising at least one enzyme selected from an alkaline cellulase and/or an enzyme having mannanase activity or the dishwashing composition according to the invention; and   b) Rinsing the dishes with water optionally comprising a rinsing aid;
 
wherein the item is a dishware or a hard surface.
       

     In one embodiment of the invention the method is hand dishwashing or automatic dish washing. In one embodiment the method is for washing or cleaning of dishware. 
     The item can be a dish ware or a hard surface. In one embodiment of the invention the item is the interior of a dishwashing machine or a sink, such as walls, baskets, nozzles, pumps, sump, filters, pipelines, drains, and outlets. 
     The at least one enzyme for preventing, reducing or removing odor from an item can be an alkaline cellulase, which alkaline cellulase is an enzyme exhibiting endo-beta-1,4-glucanase activity (EC 3.2.1.4). One example of an enzyme exhibiting endo-beta-1,4-glucanase activity is the enzyme of SEQ ID NO: 1, or an enzyme variant having at least 80% sequence identity to SEQ ID NO: 1. In one embodiment the enzyme variant has at least 85% sequence identity to SEQ ID NO: 1, at least 90% sequence identity to SEQ ID NO: 1, at least 95% sequence identity to SEQ ID NO: 1, at least 96% sequence identity to SEQ ID NO: 1, at least 97% sequence identity to SEQ ID NO: 1, at least 98% sequence identity to SEQ ID NO: 1 or at least 99% sequence identity to SEQ ID NO: 1. 
     The at least one enzyme for preventing, reducing or removing odor from an item can be a mannanase enzyme having at least 80% sequence identity to SEQ ID NO: 2. One example of an enzyme exhibiting mannanase activity is the enzyme of SEQ ID NO: 2, or an enzyme variant having at least 80% sequence identity to SEQ ID NO: 2. In one embodiment the enzyme variant has at least 85% sequence identity to SEQ ID NO: 2, at least 90% sequence identity to SEQ ID NO: 2, at least 95% sequence identity to SEQ ID NO: 2, at least 96% sequence identity to SEQ ID NO: 2, at least 97% sequence identity to SEQ ID NO: 2, at least 98% sequence identity to SEQ ID NO: 2 or at least 99% sequence identity to SEQ ID NO: 2. 
     Experiments have shown that use an alkaline cellulase and/or an enzyme having mannanase activity together with an oxidase shows even better results for preventing, reducing or removing odor from an item. In one embodiment of the invention the at least one enzyme is used together with an oxidase. 
     In one embodiment of the invention the oxidase is a carbohydrate oxidase (EC 1.1.3). The carbohydrate oxidase can be a glucose oxidase (EC 1.1.3.4), a dehydrogenase, a cellobiose oxidase (EC 1.1.99.18). 
     In one embodiment of the invention the glucose oxidase has at least 80% sequence identity to SEQ ID NO: 3. One example of an enzyme exhibiting glucose oxidase activity is the enzyme of SEQ ID NO: 3, or an enzyme variant having at least 80% sequence identity to SEQ ID NO: 3. In one embodiment the enzyme variant has at least 85% sequence identity to SEQ ID NO: 3, at least 90% sequence identity to SEQ ID NO: 3, at least 95% sequence identity to SEQ ID NO: 3, at least 96% sequence identity to SEQ ID NO: 3, at least 97% sequence identity to SEQ ID NO: 3, at least 98% sequence identity to SEQ ID NO: 3 or at least 99% sequence identity to SEQ ID NO: 3. 
     In one embodiment of the invention the cellobiose oxidase has at least 80% sequence identity to SEQ ID NO: 4. One example of an enzyme exhibiting cellobiose oxidase activity is the enzyme of SEQ ID NO: 4, or an enzyme variant having at least 80% sequence identity to SEQ ID NO: 4. In one embodiment the enzyme variant has at least 85% sequence identity to SEQ ID NO: 4, at least 90% sequence identity to SEQ ID NO: 4, at least 95% sequence identity to SEQ ID NO: 4, at least 96% sequence identity to SEQ ID NO: 4, at least 97% sequence identity to SEQ ID NO: 4, at least 98% sequence identity to SEQ ID NO: 4 or at least 99% sequence identity to SEQ ID NO: 4. 
     The alkaline cellulase and/or an enzyme having mannanase activity optionally together with an oxidase is very effectively in preventing, reducing or removing the odor for items. In one embodiment of the invention the odor is reduced by at least 50% when measured with Assay I. In one embodiment the odor is reduced by at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% when measured with Assay I. 
     The reduction in volatile compounds can also be measured by the human nose, e.g. by scoring the presence of odorous compounds according to Assay II. In one embodiment of the invention the presence of odor is scored on average 3 points lower than control (without use of the inventive enzymes) when measured with Assay II. In one embodiment the presence of odor is scored on average 4 points, 5 points, 6 points, 7 points or 8 points lower than control when measured with Assay II. 
     The alkaline cellulase and/or an enzyme having mannanase activity optionally together with an oxidase can be used together with other enzymes. The enzymes may be used together with one or more enzymes such as a protease, lipase, cutinase, an amylase, carbohydrase, cellulase, pectinase, mannanase, arabinase, galactanase, xylanase, DNase, oxidase, e.g., a laccase, and/or peroxidase. 
     In one embodiment of the invention the alkaline cellulase and/or an enzyme having mannanase activity optionally together with an oxidase may be used with an amylase and/or a protease. 
     In one embodiment of the invention the amylase is an alpha-amylase or a glucoamylase. The amylase may be of bacterial or fungal origin. In one embodiment the amylase is an alpha-amylase obtained from  Bacillus , such as  Bacillus licheniformis.    
     In one embodiment of the invention the protease is chemically modified or protein engineered. The protease can be a serine protease or a metalloprotease, preferably an alkaline microbial protease or a trypsin-like protease. In one embodiment the protease is selected from the group consisting of  Bacillus , e.g., subtilisin Novo, subtilisin Carlsberg, subtilisin 309, subtilisin 147, subtilisin 168, trypsin of bovine origin, trypsin of porcine origin and  Fusarium  protease. 
     When the alkaline cellulase and/or an enzyme having mannanase activity is present in a dishwashing composition the composition may further comprise a builder. The composition may further comprise a surfactant. In one embodiment of the invention the composition further comprises one or more builders and one or more polymer. 
     The composition may further comprise one or more components selected from the group consisting of polymers, bleaching systems, bleach activators, bleach catalysts, silicates, dyestuff and metal care agents. The components are further described in the below paragraphs. 
     The composition can be in the form of a powder, a bar, a homogenous tablet, a tablet having two or more layers, a pouch having one or more compartments, a regular or compact powder, a granule, a paste, a gel, or a regular, compact or concentrated liquid. 
     Surfactants 
     The dish washing composition can include at least one non-ionic surfactant. Suitable nonionic surfactants include, but are not limited to low-foaming nonionic (LFNI) surfactants. A LFNI surfactant is most typically used in an automatic dishwashing composition because of the improved water-sheeting action (especially from glassware) which they confer to the automatic dishwashing composition. They also may encompass non-silicone, phosphate or nonphosphate polymeric materials which are known to defoam food soils encountered in automatic dishwashing. The LFNI surfactant may have a relatively low cloud point and a high hydrophilic-lipophilic balance (HLB). Cloud points of 1% solutions in water are typically below about 32° C. and alternatively lower, e.g., 0° C., for optimum control of sudsing throughout a full range of water temperatures. If desired, a biodegradable LFNI surfactant having the above properties may be used. 
     A LFNI surfactant may include, but is not limited to: alkoxylated surfactants, especially ethoxylates derived from primary alcohols, and blends thereof with more sophisticated surfactants, such as the polyoxypropylene/polyoxyethylene/polyoxypropylene reverse block polymers. Suitable block polyoxyethylene-polyoxypropylene polymeric compounds that meet the requirements may include those based on ethylene glycol, propylene glycol, glycerol, trimethylolpropane and ethylenediamine, and mixtures thereof. Polymeric compounds made from a sequential ethoxylation and propoxylation of initiator compounds with a single reactive hydrogen atom, such as C 12—is aliphatic alcohols, do not generally provide satisfactory suds control in Automatic dishwashing compositions. However, certain of the block polymer surfactant compounds designated as PLURONIC® and TETRONIC® by the BASF-Wyandotte Corp., Wyandotte, Mich., are suitable in Automatic dishwashing compositions. The LFNI surfactant can optionally include a propylene oxide in an amount up to about 15% by weight. Other LFNI surfactants can be prepared by the processes described in U.S. Pat. No. 4,223,163. The LFNI surfactant may also be derived from a straight chain fatty alcohol containing from about 16 to about 20 carbon atoms (C16-C20 alcohol), alternatively a Cl8 alcohol, condensed with an average of from about 6 to about 15 moles, or from about 7 to about 12 moles, and alternatively, from about 7 to about 9 moles of ethylene oxide per mole of alcohol. The ethoxylated nonionic surfactant so derived may have a narrow ethoxylate distribution relative to the average. 
     In certain embodiments, a LFNI surfactant having a cloud point below 30° C. may be present in an amount from about 0.01% to about 60%, or from about 0.5% to about 10% by weight, and alternatively, from about 1% to about 5% by weight of the composition 
     In preferred embodiments, the surfactant is a non-ionic surfactant or a non-ionic surfactant system having a phase inversion temperature, as measured at a concentration of 1% in distilled water, between 40 and 70° C., preferably between 45 and 65° C. By a “non-ionic surfactant system” is meant herein a mixture of two or more non-ionic surfactants. Preferred for use herein are non-ionic surfactant systems. They seem to have improved cleaning and finishing properties and stability in product than single non-ionic surfactants. Suitable nonionic surfactants include: i) ethoxylated non-ionic surfactants prepared by the reaction of a monohydroxy alkanol or alkyphenol with 6 to 20 carbon atoms with preferably at least 12 moles particularly preferred at least 16 moles, and still more preferred at least 20 moles of ethylene oxide per mole of alcohol or alkylphenol; ii) alcohol alkoxylated surfactants having a from 6 to 20 carbon atoms and at least one ethoxy and propoxy group. Preferred for use herein are mixtures of surfactants i) and ii). 
     Another suitable non-ionic surfactants are epoxy-capped poly(oxyalkylated) alcohols represented by the formula: 
       R 1 O[CH 2 CH(CH 3 )O] x [CH 2 CH 2 O] y [CH 2 CH(OH)R 2 ]  (I)
 
     wherein R 1  is a linear or branched, aliphatic hydrocarbon radical having from 4 to 18 carbon atoms; R 2  is a linear or branched aliphatic hydrocarbon radical having from 2 to 26 carbon atoms; x is an integer having an average value of from 0.5 to 1.5, more preferably about 1; and y is an integer having a value of at least 15, more preferably at least 20. Preferably, the surfactant of formula I has at least about 10 carbon atoms in the terminal epoxide unit [CH 2 CH(OH)R 2 ]. Suitable surfactants of formula I are Olin Corporation&#39;s POLY-TERGENT® SLF-18B nonionic surfactants, as described, for example, in WO 94/22800, published Oct. 13, 1994 by Olin Corporation. 
     Preferably non-ionic surfactants and/or system herein have a Draves wetting time of less than 360 seconds, preferably less than 200 seconds, more preferably less than 100 seconds and especially less than 60 seconds as measured by the Draves wetting method (standard method ISO 8022 using the following conditions; 3-g hook, 5-g cotton skein, 0.1% by weight aqueous solution at a temperature of 25° C.). Amine oxides surfactants are also useful in the present invention as anti-redeposition surfactants include linear and branched compounds having the formula: 
     
       
         
         
             
             
         
       
     
     wherein R 3  is selected from an alkyl, hydroxyalkyl, acylamidopropoyl and alkyl phenyl group, or mixtures thereof, containing from 8 to 26 carbon atoms, preferably 8 to 18 carbon atoms; R 4  is an alkylene or hydroxyalkylene group containing from 2 to 3 carbon atoms, preferably 2 carbon atoms, or mixtures thereof; x is from 0 to 5, preferably from 0 to 3; and each R 5  is an alkyl or hydroxyalkyl group containing from 1 to 3, preferably from 1 to 2 carbon atoms, or a polyethylene oxide group containing from 1 to 3, preferable 1, 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 18  alkyl dimethyl amine oxides and C 8 -C 18  alkoxy ethyl dihydroxyethyl amine oxides. Examples of such materials include dimethyloctylamine oxide, diethyldecylamine oxide, bis-(2-hydroxyethyl)dodecylamine oxide, dimethyldodecylamine oxide, dipropyltetradecylamine oxide, methylethylhexadecylamine oxide, dodecylamidopropyl dimethylamine oxide, cetyl dimethylamine oxide, stearyl dimethylamine oxide, tallow dimethylamine oxide and dimethyl-2-hydroxyoctadecylamine oxide. Preferred are C 10 -C 18  alkyl dimethylamine oxide, and C 10 -C 18  acylamido alkyl dimethylamine oxide. Surfactants and especially non-ionic surfactants may be present in amounts from 0 to 10% by weight, preferably from 0.1% to 10%, and most preferably from 0.25% to 6%. 
     Sulfonated Polymer 
     The polymer, if used, is used in any suitable amount from about 0.1% to about 20%, preferably from 1% to about 15%, more preferably from 2% to 10% by weight of the composition. Sulfonated/carboxylated polymers are particularly suitable for the compositions contained in the pouch of the invention. 
     Suitable sulfonated/carboxylated polymers described herein may have a weight average molecular weight of less than or equal to about 100,000 Da, or less than or equal to about 75,000 Da, or less than or equal to about 50,000 Da, or from about 3,000 Da to about 50,000, preferably from about 5,000 Da to about 45,000 Da. 
     As noted herein, the sulfonated/carboxylated polymers may comprise (a) at least one structural unit derived from at least one carboxylic acid monomer having the general formula (I): 
     
       
         
         
             
             
         
       
     
     wherein R 1  to R 4  are independently hydrogen, methyl, carboxylic acid group or CH 2 COOH and wherein the carboxylic acid groups can be neutralized; (b) optionally, one or more structural units derived from at least one nonionic monomer having the general formula (II): 
     
       
         
         
             
             
         
       
     
     wherein R 5  i is hydrogen, C 1  to C 6  alkyl, or C 1  to C 6  hydroxyalkyl, and X is either aromatic (with R 5  being hydrogen or methyl when X is aromatic) or X is of the general formula (III): 
     
       
         
         
             
             
         
       
     
     wherein R 6  is (independently of R 5 ) hydrogen, C 1  to C 6  alkyl, or C 1  to C 6  hydroxyalkyl, and Y is O or N; and at least one structural unit derived from at least one sulfonic acid monomer having the general formula (IV): 
     
       
         
         
             
             
         
       
     
     wherein R 7  is a group comprising at least one sp 2  bond, A is O, N, P, S or an amido or ester linkage, B is a mono- or polycyclic aromatic group or an aliphatic group, each t is independently 0 or 1, and M +  is a cation. In one aspect, R 7  is a C 2  to C 6  alkene. In another aspect, R 7  is ethene, butene or propene. 
     Preferred carboxylic acid monomers include one or more of the following: acrylic acid, maleic acid, itaconic acid, methacrylic acid, or ethoxylate esters of acrylic acids, acrylic and methacrylic acids being more preferred. Preferred sulfonated monomers include one or more of the following: sodium (meth) allyl sulfonate, vinyl sulfonate, sodium phenyl (meth) allyl ether sulfonate, or 2-acrylamido-methyl propane sulfonic acid. Preferred non-ionic monomers include one or more of the following: methyl (meth) acrylate, ethyl (meth) acrylate, t-butyl (meth) acrylate, methyl (meth) acrylamide, ethyl (meth) acrylamide, t-butyl (meth) acrylamide, styrene, or [alpha]-methyl styrene. 
     Preferably, the polymer comprises the following levels of monomers: from about 40 to about 90%, preferably from about 60 to about 90% by weight of the polymer of one or more carboxylic acid monomer; from about 5 to about 50%, preferably from about 10 to about 40% by weight of the polymer of one or more sulfonic acid monomer; and optionally from about 1% to about 30%, preferably from about 2 to about 20% by weight of the polymer of one or more non-ionic monomer. An especially preferred polymer comprises about 70% to about 80% by weight of the polymer of at least one carboxylic acid monomer and from about 20% to about 30% by weight of the polymer of at least one sulfonic acid monomer. 99 The carboxylic acid is preferably (meth)acrylic acid. The sulfonic acid monomer is preferably one of the following: 2-acrylamido methyl-1-propanesulfonic acid, 2-methacrylamido-2-methyl-1-propanesulfonic acid, 3-methacrylamido-2-hydroxypropanesulfonic acid, allysulfonic acid, methallysulfonic acid, allyloxybenzenesulfonic acid, methallyloxybenzensulfonic acid, 2-hydroxy-3-(2-propenyloxy)propanesulfonic acid, 2-methyl-2-propene-I-sulfonic acid, styrene sulfonic acid, vinylsulfonic acid, 3-sulfopropyl acrylate, 3-sulfopropyl methacrylate, sulfomethylacrylamid, sulfomethylmethacrylamide, and water soluble salts thereof. The unsaturated sulfonic acid monomer is most preferably 2-acrylamido-2-propanesulfonic acid (AMPS). 
     Preferred commercial available polymers include: Alcosperse 240, Aquatreat AR 540 and Aquatreat MPS supplied by Alco Chemical; Acumer 3100, Acumer 2000, Acusol 587G and Acusol 588G supplied by Rohm &amp; Haas; Goodrich K-798, K-775 and K-797 supplied by BF Goodrich; and ACP 1042 supplied by ISP technologies Inc. Particularly preferred polymers are Acusol 587G and Acusol 588G supplied by Rohm &amp; Haas. 
     In the polymers, all or some of the carboxylic or sulfonic acid groups can be present in neutralized form, i.e. the acidic hydrogen atom of the carboxylic and/or sulfonic acid group in some or all acid groups can be replaced with metal ions, preferably alkali metal ions and in particular with sodium ions. 
     Hydrotropes 
     A hydrotrope is a compound that solubilises hydrophobic compounds in aqueous solutions (or oppositely, polar substances in a non-polar environment). Typically, hydrotropes have both hydrophilic and a hydrophobic character (so-called amphiphilic properties as known from surfactants); however the molecular structure of hydrotropes generally do not favor spontaneous self-aggregation, see e.g. review by Hodgdon and Kaler (2007), Current Opinion in Colloid &amp; Interface Science 12: 121-128. Hydrotropes do not display a critical concentration above which self-aggregation occurs as found for surfactants and lipids forming miceller, lamellar or other well defined meso-phases. Instead, many hydrotropes show a continuous-type aggregation process where the sizes of aggregates grow as concentration increases. However, many hydrotropes alter the phase behavior, stability, and colloidal properties of systems containing substances of polar and non-polar character, including mixtures of water, oil, surfactants, and polymers. Hydrotropes are classically used across industries from pharma, personal care, food, to technical applications. Use of hydrotropes in detergent compositions allow for example more concentrated formulations of surfactants (as in the process of compacting liquid detergents by removing water) without inducing undesired phenomena such as phase separation or high viscosity. 
     The detergent may contain 0-10% by weight, for example 0-5% by weight, such as about 0.5 to about 5%, or about 3% to about 5%, of a hydrotrope. Any hydrotrope known in the art for use in detergents may be utilized. Non-limiting examples of hydrotropes include sodium benzenesulfonate, sodium p-toluene sulfonate (STS), sodium xylene sulfonate (SXS), sodium cumene sulfonate (SCS), sodium cymene sulfonate, amine oxides, alcohols and polyglycolethers, sodium hydroxynaphthoate, sodium hydroxynaphthalene sulfonate, sodium ethylhexyl sulfate, and combinations thereof. 
     Builders and Co-Builders 
     The detergent composition may contain about 0-65% by weight, such as about 5% to about 50% of a detergent builder or co-builder, or a mixture thereof. In a dish wash detergent, the level of builder is typically 40-65%, particularly 50-65%. The builder and/or co-builder may particularly be a chelating agent that forms water-soluble complexes with Ca and Mg. Any builder and/or co-builder known in the art for use in ADW detergents may be utilized. Non-limiting examples of builders include zeolites, diphosphates (pyrophosphates), triphosphates such as sodium triphosphate (STP or STPP), carbonates such as sodium carbonate, soluble silicates such as sodium metasilicate, layered silicates (e.g., SKS-6 from Hoechst), ethanolamines such as 2-aminoethan-1-ol (MEA), diethanolamine (DEA, also known as 2,2′-iminodiethan-1-ol), triethanolamine (TEA, also known as 2,2′,2″-nitrilotriethan-1-ol), and (carboxymethyl)inulin (CMI), and combinations thereof. 
     The detergent composition may also contain 0-50% by weight, such as about 5% to about 30%, of a detergent co-builder. The detergent composition may include include a co-builder alone, or in combination with a builder, for example a zeolite builder. Non-limiting examples of co-builders include homopolymers of polyacrylates or copolymers thereof, such as poly(acrylic acid) (PAA) or copoly(acrylic acid/maleic acid) (PAA/PMA). Further non-limiting examples include citrate, chelators such as aminocarboxylates, aminopolycarboxylates and phosphonates, and alkyl- or alkenylsuccinic acid. Additional specific examples include 2,2′,2″-nitrilotriacetic acid (NTA), ethylenediaminetetraacetic acid (EDTA), diethylenetriaminepentaacetic acid (DTPA), iminodisuccinic acid (IDS), ethylenediamine-N,N′-disuccinic acid (EDDS), methylglycinediacetic acid (MGDA), glutamic acid-N,N-diacetic acid (GLDA), 1-hydroxyethane-1,1-diphosphonic acid (HEDP), ethylenediaminetetra(methylenephosphonic acid) (EDTMPA), diethylenetriaminepentakis(methylenephosphonic acid) (DTMPA or DTPMPA), N-(2-hydroxyethyl)iminodiacetic acid (EDG), aspartic acid-N-monoacetic acid (ASMA), aspartic acid-N,N-diacetic acid (ASDA), aspartic acid-N-monopropionic acid (ASMP), iminodisuccinic acid (IDA), N-(2-sulfomethyl)-aspartic acid (SMAS), N-(2-sulfoethyl)-aspartic acid (SEAS), N-(2-sulfomethyl)-glutamic acid (SMGL), N-(2-sulfoethyl)-glutamic acid (SEGL), N-methyliminodiacetic acid (MIDA), α-alanine-N,N-diacetic acid (α-ALDA), serine-N,N-diacetic acid (SEDA), isoserine-N,N-diacetic acid (ISDA), phenylalanine-N,N-diacetic acid (PHDA), anthranilic acid-N,N-diacetic acid (ANDA), sulfanilic acid-N,N-diacetic acid (SLDA), taurine-N,N-diacetic acid (TUDA) and sulfomethyl-N,N-diacetic acid (SMDA), N-(2-hydroxyethyl)ethylenediamine-N,N,N″-triacetic acid (HEDTA), diethanolglycine (DEG), diethylenetriamine penta(methylenephosphonic acid) (DTPMP), aminotris(methylenephosphonic acid) (ATMP), and combinations and salts thereof. Further exemplary builders and/or co-builders are described in, e.g., WO 09/102854, U.S. Pat. No. 5,977,053 
     The dish wash composition contains 0-50% by weight, such as about 5% to about 30%, of a detergent co-builder and/or the dish wash compositions is preferably phosphate free. According to one aspect of the invention the builder is selected from citric acid, methyl glycine-N,N-diacetic acid (MGDA) and/or glutamic-N,N-diacetic acid (GLDA) and mixtures thereof. According to one aspect of the invention the bleach component is selected from bleaching catalysts, photobleaches, bleach activators, sources of hydrogen peroxide such as sodium percarbonate, sodium perborates and hydrogen peroxide, preformed peracids and mixtures thereof. According to one aspect the bleach component is a peroxide such as percarbonate, persulfate, perphosphate, persilicate salts. In one aspect the bleaching component includes a percarbonate and bleach catalyst, preferably a manganese compound according to one aspect the bleach catalyst is 1,4,7-trimethyl-1,4,7-triazacyclononane or manganese (II) acetate tetrahydrate (MnTACN). In one aspect the dish wash composition comprising from 1-40 wt %, preferably from 0.5-30 wt %, of bleaching components, wherein the bleach components are a peroxide, preferably percabonate and a catalyst preferably a metal-containing bleach catalyst such as 1,4,7-trimethyl-1,4,7-triazacyclononane or manganese (II) acetate tetrahydrate (MnTACN). 
     Bleaching Systems 
     Inorganic and organic bleaches are suitable cleaning actives for use herein. Inorganic bleaches include perhydrate salts such as perborate, percarbonate, perphosphate, persulfate and persilicate salts. The inorganic perhydrate salts are normally the alkali metal salts. The inorganic perhydrate salt may be included as the crystalline solid without additional protection. Alternatively, the salt can be coated. 
     Alkali metal percarbonates, particularly sodium percarbonate are preferred perhydrates for use herein. The percarbonate is most preferably incorporated into the products in a coated form which provides in-product stability. A suitable coating material providing in product stability comprises mixed salt of a water-soluble alkali metal sulphate and carbonate. Such coatings together with coating processes have previously been described in GB-1,466,799. The weight ratio of the mixed salt coating material to percarbonate lies in the range from 1:200 to 1:4, more preferably from 1:99 to 19, and most preferably from 1:49 to 1:19. Preferably, the mixed salt is of sodium sulphate and sodium carbonate which has the general formula Na2SO4.n.Na2CO3 wherein n is from 0.1 to 3, preferably n is from 0.3 to 1.0 and most preferably n is from 0.2 to 0.5. 
     Another suitable coating material providing in product stability, comprises sodium silicate of SiO 2 :Na2O ratio from 1.8:1 to 3.0:1, preferably L8:1 to 2.4:1, and/or sodium metasilicate, preferably applied at a level of from 2% to 10%, (normally from 3% to 5%) of SiO 2  by weight of the inorganic perhydrate salt. Magnesium silicate can also be included in the coating. Coatings that contain silicate and borate salts or boric acids or other inorganics are also suitable. 
     Other coatings which contain waxes, oils, fatty soaps can also be used advantageously within the present invention. 
     Potassium peroxymonopersulfate is another inorganic perhydrate salt of utility herein. Typical organic bleaches are organic peroxyacids including diacyl and tetraacylperoxides, especially diperoxydodecanedioc acid, diperoxytetradecanedioc acid, and diperoxyhexadecanedioc acid. Dibenzoyl peroxide is a preferred organic peroxyacid herein. Mono- and diperazelaic acid, mono- and diperbrassylic acid, and Nphthaloylaminoperoxicaproic acid are also suitable herein. The diacyl peroxide, especially dibenzoyl peroxide, should preferably be present in the form of particles having a weight average diameter of from about 0.1 to about 100 microns, preferably from about 0.5 to about 30 microns, more preferably from about 1 to about 10 microns. Preferably, at least about 25%, more preferably at least about 50%, even more preferably at least about 75%, most preferably at least about 90%, of the particles are smaller than 10 microns, preferably smaller than 6 microns. Diacyl peroxides within the above particle size range have also been found to provide better stain removal especially from plastic dishware, while minimizing undesirable deposition and filming during use in automatic dishwashing machines, than larger diacyl peroxide particles. The preferred diacyl peroxide particle size thus allows the formulator to obtain good stain removal with a low level of diacyl peroxide, which reduces deposition and filming. Conversely, as diacyl peroxide particle size increases, more diacyl peroxide is needed for good stain removal, which increases deposition on surfaces encountered during the dishwashing process. Further typical organic bleaches include the peroxy acids, particular examples being the alkylperoxy acids and the arylperoxy acids. Preferred representatives are (a) peroxybenzoic acid and its ring-substituted derivatives, such as alkylperoxybenzoic acids, but also peroxy-[alpha]-naphthoic acid and magnesium monoperphthalate, (b) the aliphatic or substituted aliphatic peroxy acids, such as peroxylauric acid, peroxystearic acid, [epsilon]-phthalimidoperoxycaproic acid[phthaloiminoperoxyhexanoic acid (PAP)], o-carboxybenzamidoperoxycaproic acid, N-nonenylamidoperadipic acid and N-nonenylamidopersuccinates, and (c) aliphatic and araliphatic peroxydicarboxylic acids, such as 1,12-diperoxycarboxylic acid, 1,9-diperoxyazelaic acid, diperoxysebacic acid, diperoxybrassylic acid, the diperoxyphthalic acids, 2-decyldiperoxybutane-1,4-dioic acid, N,N-terephthaloyldi(6-aminopercaproic acid). 
     Bleach Activators 
     Bleach activators are typically organic peracid precursors that enhance the bleaching action in the course of cleaning at temperatures of 60[deg.] C. and below. Bleach activators suitable for use herein include compounds which, under perhydrolysis conditions, give aliphatic peroxoycarboxylic acids having preferably from 1 to 10 carbon atoms, in particular from 2 to 4 carbon atoms, and/or optionally substituted perbenzoic acid. Suitable substances bear O-acyl and/or N-acyl groups of the number of carbon atoms specified and/or optionally substituted benzoyl groups. Preference is given to polyacylated alkylenediamines, in particular tetraacetylethylenediamine (TAED), acylated triazine derivatives, in particular 1,5-diacetyl-2,4-dioxohexahydro-1,3,5-triazine (DADHT), acylated glycolurils, in particular tetraacetylglycoluril (TAGU), N-acylimides, in particular N-nonanoylsuccinimide (NOSI), acylated phenolsulfonates, in particular n-nonanoyl- or isononanoyloxybenzenesulfonate (n- or iso-NOBS), carboxylic anhydrides, in particular phthalic anhydride, acylated polyhydric alcohols, in particular triacetin, ethylene glycol diacetate and 2,5-diacetoxy-2,5-dihydrofuran and also triethylacetyl citrate (TEAC). Bleach activators if included in the compositions of the invention are in a level of from about 0.1 to about 10%, preferably from about 0.5 to about 2% by weight of the composition. 
     Bleach Catalysts and Bleach Boosters 
     The bleaching system may also include a bleach catalyst or booster. 
     Bleach catalysts preferred for use herein include the manganese triazacyclononane, MnTACN and related complexes (U.S. Pat. Nos. 4,246,612, 5,227,084); Co, Cu, Mn and Fe bispyridylamine and related complexes (U.S. Pat. No. 5,114,611); and pentamine acetate cobalt(III) and related complexes(U.S. Pat. No. 4,810,410). A complete description of bleach catalysts suitable for use herein can be found in WO 99/06521, pages 34, line 26 to page 40, line 16. Bleach catalyst if included in the compositions of the invention are in a level of from about 0.1 to about 10%, preferably from about 0.5 to about 2% by weight of the composition. Oxidoreductases, for example oxidases, oxygenases, catalases, peroxidases such as halo-, chloro-, bromo-, lignin, glucose, or manganese peroxidases, dioxygenases, or laccases (phenoloxidases, polyphenoloxidases), can also be used according to the present invention to intensify the bleaching effect. Advantageously, preferably organic, particularly preferably aromatic compounds that interact with the enzymes are additionally added in order to enhance the activity of the relevant oxidoreductases (enhancers) or, if there is a large difference in redox potentials between the oxidizing enzymes and the stains, to ensure electron flow (mediators). 
     Some non-limiting examples of bleach catalysts that may be used in the compositions of the present invention include manganese oxalate, manganese acetate, manganese-collagen, cobalt-amine catalysts and manganese triazacyclononane (MnTACN) catalysts; particularly preferred are complexes of manganese with 1,4,7-trimethyl-1,4,7-triazacyclononane (Me3-TACN) or 1,2,4,7-tetramethyl-1,4,7-triazacyclononane (Me4-TACN), in particular Me3-TACN, such as the dinuclear manganese complex [(Me3-TACN)Mn(O)3Mn(Me3-TACN)](PF6)2, and [2,2′,2″-nitrilotris(ethane-1,2-diylazanylylidene-κN-methanylylidene)triphenolato-κ3O]manganese(111). The bleach catalysts may also be other metal compounds, such as iron or cobalt complexes. 
     In some embodiments, where a source of a peracid is included, an organic bleach catalyst or bleach booster may be used having one of the following formulae: 
     
       
         
         
             
             
         
       
     
     (iii) and mixtures thereof; wherein each R1 is independently a branched alkyl group containing from 9 to 24 carbons or linear alkyl group containing from 11 to 24 carbons, preferably each R1 is independently a branched alkyl group containing from 9 to 18 carbons or linear alkyl group containing from 11 to 18 carbons, more preferably each R1 is independently selected from the group consisting of 2-propylheptyl, 2-butyloctyl, 2-pentylnonyl, 2-hexyldecyl, dodecyl, tetradecyl, hexadecyl, octadecyl, isononyl, isodecyl, isotridecyl and isopentadecyl.
 
Other exemplary bleaching systems are described, e.g. in WO2007/087258, WO2007/087244, WO2007/087259, EP1867708 (Vitamin K) and WO2007/087242. Suitable photobleaches may for example be sulfonated zinc or aluminium phthalocyanines.
 
Other exemplary bleaching systems are described, e.g. in WO2007/087258, WO2007/087244, WO2007/087259, EP1867708 (Vitamin K) and WO2007/087242. Suitable photobleaches may for example be sulfonated zinc or aluminium phthalocyanines.
 
     Silicates 
     Preferred silicates are sodium silicates such as sodium disilicate, sodium metasilicate and crystalline phyllosilicates. Silicates if present are at a level of from about 1 to about 20%, preferably from about 5 to about 15% by weight of composition. 
     Metal Care Agents 
     Metal care agents may prevent or reduce the tarnishing, corrosion or oxidation of metals, including aluminium, stainless steel and non-ferrous metals, such as silver and copper. Suitable examples include one or more of the following: 
     (a) benzatriazoles, including benzotriazole or bis-benzotriazole and substituted derivatives thereof. Benzotriazole derivatives are those compounds in which the available substitution sites on the aromatic ring are partially or completely substituted. Suitable substituents include linear or branch-chain Ci-C20-alkyl groups and hydroxyl, thio, phenyl or halogen such as fluorine, chlorine, bromine and iodine. 
     (b) metal salts and complexes chosen from the group consisting of zinc, manganese, titanium, zirconium, hafnium, vanadium, cobalt, gallium and cerium salts and/or complexes, the metals being in one of the oxidation states II, III, IV, V or VI. In one aspect, suitable metal salts and/or metal complexes may be chosen from the group consisting of Mn(II) sulphate, Mn(II) citrate, Mn(II) stearate, Mn(II) acetylacetonate, K̂TiF6, K̂ZrF6, CoSO4, Co(NOs)2 and Ce(NOs)3, zinc salts, for example zinc sulphate, hydrozincite or zinc acetate; (c) silicates, including sodium or potassium silicate, sodium disilicate, sodium metasilicate, crystalline phyllosilicate and mixtures thereof. 
     Further suitable organic and inorganic redox-active substances that act as silver/copper corrosion inhibitors are disclosed in WO 94/26860 and WO 94/26859. Preferably the composition of the invention comprises from 0.1 to 5% by weight of the composition of a metal care agent, preferably the metal care agent is a zinc salt. 
     Polymers 
     The detergent may contain 0-10% by weight, such as 0.5-5%, 2-5%, 0.5-2% or 0.2-1% of a polymer. Any polymer known in the art for use in detergents may be utilized. The polymer may function as a co-builder as mentioned above, or may provide antiredeposition, fiber protection, soil release, dye transfer inhibition, grease cleaning and/or anti-foaming properties. Some polymers may have more than one of the above-mentioned properties and/or more than one of the below-mentioned motifs. Exemplary polymers include (carboxymethyl)cellulose (CMC), poly(vinyl alcohol) (PVA), poly(vinylpyrrolidone) (PVP), poly(ethyleneglycol) or poly(ethylene oxide) (PEG), ethoxylated poly(ethyleneimine), carboxymethyl inulin (CMI), and polycarboxylates such as PAA, PAA/PMA, poly-aspartic acid, and lauryl methacrylate/acrylic acid copolymers, hydrophobically modified CMC (HM-CMC) and silicones, copolymers of terephthalic acid and oligomeric glycols, copolymers of poly(ethylene terephthalate) and poly(oxyethene terephthalate) (PET-POET), PVP, poly(vinylimidazole) (PVI), poly(vinylpyridine-N-oxide) (PVPO or PVPNO) and polyvinylpyrrolidone-vinylimidazole (PVPVI). Further exemplary polymers include sulfonated polycarboxylates, polyethylene oxide and polypropylene oxide (PEO-PPO) and diquaternium ethoxy sulfate. Other exemplary polymers are disclosed in, e.g., WO 2006/130575. Salts of the above-mentioned polymers are also contemplated. 
     Adjunct Materials 
     Any detergent components known in the art for use in ADW detergents may also be utilized. Other optional detergent components include anti-corrosion agents, anti-shrink agents, anti-soil redeposition agents, anti-wrinkling agents, bactericides, binders, corrosion inhibitors, disintegrants/disintegration agents, dyes, enzyme stabilizers (including boric acid, borates, CMC, and/or polyols such as propylene glycol), fabric conditioners including clays, fillers/processing aids, fluorescent whitening agents/optical brighteners, foam boosters, foam (suds) regulators, perfumes, soil-suspending agents, softeners, suds suppressors, tarnish inhibitors, and wicking agents, either alone or in combination. Any ingredient known in the art for use ADW detergents may be utilized. The choice of such ingredients is well within the skill of the artisan. 
     Dispersants 
     The detergent compositions of the present invention can also contain dispersants. In particular powdered detergents may comprise dispersants. Suitable water-soluble organic materials include the homo- or co-polymeric acids or their salts, in which the polycarboxylic acid comprises at least two carboxyl radicals separated from each other by not more than two carbon atoms. Suitable dispersants are for example described in Powdered Detergents, Surfactant science series volume 71, Marcel Dekker, Inc. 
     Dye Transfer Inhibiting Agents 
     The detergent compositions of the present invention may also include one or more dye transfer inhibiting agents. Suitable polymeric dye transfer inhibiting agents include, but are not limited to, polyvinylpyrrolidone polymers, polyamine N-oxide polymers, copolymers of N-vinylpyrrolidone and N-vinylimidazole, polyvinyloxazolidones and polyvinylimidazoles or mixtures thereof. When present in a subject composition, the dye transfer inhibiting agents may be present at levels from about 0.0001% to about 10%, from about 0.01% to about 5% or even from about 0.1% to about 3% by weight of the composition. 
     Fluorescent Whitening Agent 
     The detergent compositions of the present invention will preferably also contain additional components that may tint articles being cleaned, such as fluorescent whitening agent or optical brighteners. Where present the brightener is preferably at a level of about 0.01% to about 0.5%. Any fluorescent whitening agent suitable for use in a laundry detergent composition may be used in the composition of the present invention. The most commonly used fluorescent whitening agents are those belonging to the classes of diaminostilbene-sulfonic acid derivatives, diarylpyrazoline derivatives and bisphenyl-distyryl derivatives. Examples of the diaminostilbene-sulfonic acid derivative type of fluorescent whitening agents include the sodium salts of: 4,4′-bis-(2-diethanolamino-4-anilino-s-triazin-6-ylamino) stilbene-2,2′-disulfonate, 4,4′-bis-(2,4-dianilino-s-triazin-6-ylamino) stilbene-2,2′-disulfonate, 4,4′-bis-(2-anilino-4-(N-methyl-N-2-hydroxy-ethylamino)-s-triazin-6-ylamino) stilbene-2,2′-disulfonate, 4,4′-bis-(4-phenyl-1,2,3-triazol-2-yl)stilbene-2,2′-disulfonate and sodium 5-(2H-naphtho[1,2-d][1,2,3]triazol-2-yl)-2-[(E)-2-phenylvinyl]benzenesulfonate. Preferred fluorescent whitening agents are Tinopal DMS and Tinopal CBS available from Ciba-Geigy AG, Basel, Switzerland. Tinopal DMS is the disodium salt of 4,4′-bis-(2-morpholino-4-anilino-s-triazin-6-ylamino) stilbene-2,2′-disulfonate. Tinopal CBS is the disodium salt of 2,2′-bis-(phenyl-styryl)-disulfonate. Also preferred are fluorescent whitening agents is the commercially available Parawhite KX, supplied by Paramount Minerals and Chemicals, Mumbai, India. Other fluorescers suitable for use in the invention include the 1-3-diaryl pyrazolines and the 7-alkylaminocoumarins. 
     Suitable fluorescent brightener levels include lower levels of from about 0.01, from 0.05, from about 0.1 or even from about 0.2 wt % to upper levels of 0.5 or even 0.75 wt %. 
     Soil Release Polymers 
     The detergent compositions of the present invention may also include one or more soil release polymers which aid the removal of soils from fabrics such as cotton and polyester based fabrics, in particular the removal of hydrophobic soils from polyester based fabrics. The soil release polymers may for example be nonionic or anionic terephthalte based polymers, polyvinyl caprolactam and related copolymers, vinyl graft copolymers, polyester polyamides see for example Chapter 7 in Powdered Detergents, Surfactant science series volume 71, Marcel Dekker, Inc. Another type of soil release polymers are amphiphilic alkoxylated grease cleaning polymers comprising a core structure and a plurality of alkoxylate groups attached to that core structure. The core structure may comprise a polyalkylenimine structure or a polyalkanolamine structure as described in detail in WO 2009/087523 (hereby incorporated by reference). Furthermore random graft co-polymers are suitable soil release polymers. Suitable graft co-polymers are described in more detail in WO 2007/138054, WO 2006/108856 and WO 2006/113314 (hereby incorporated by reference). Other soil release polymers are substituted polysaccharide structures especially substituted cellulosic structures such as modified cellulose deriviatives such as those described in EP 1867808 or WO 2003/040279 (both are hereby incorporated by reference). Suitable cellulosic polymers include cellulose, cellulose ethers, cellulose esters, cellulose amides and mixtures thereof. Suitable cellulosic polymers include anionically modified cellulose, nonionically modified cellulose, cationically modified cellulose, zwitterionically modified cellulose, and mixtures thereof. Suitable cellulosic polymers include methyl cellulose, carboxy methyl cellulose, ethyl cellulose, hydroxyl ethyl cellulose, hydroxyl propyl methyl cellulose, ester carboxy methyl cellulose, and mixtures thereof. 
     Anti-Redeposition Agents 
     The detergent compositions of the present invention may also include one or more anti-redeposition agents such as carboxymethylcellulose (CMC), polyvinyl alcohol (PVA), polyvinylpyrrolidone (PVP), polyoxyethylene and/or polyethyleneglycol (PEG), homopolymers of acrylic acid, copolymers of acrylic acid and maleic acid, and ethoxylated polyethyleneimines. The cellulose based polymers described under soil release polymers above may also function as anti-redeposition agents. 
     Rheology Modifiers 
     The detergent compositions of the present invention may also include one or more rheology modifiers, structurants or thickeners, as distinct from viscosity reducing agents. The rheology modifiers are selected from the group consisting of non-polymeric crystalline, hydroxy-functional materials, polymeric rheology modifiers which impart shear thinning characteristics to the aqueous liquid matrix of a liquid detergent composition. The rheology and viscosity of the detergent can be modified and adjusted by methods known in the art, for example as shown in EP 2169040. 
     Other suitable adjunct materials include, but are not limited to, anti-shrink agents, anti-wrinkling agents, bactericides, binders, carriers, dyes, enzyme stabilizers, fabric softeners, fillers, foam regulators, hydrotropes, perfumes, pigments, sod suppressors, solvents, and structurants for liquid detergents and/or structure elasticizing agents. 
     Formulation of Detergent Products 
     The detergent composition of the invention may be in any convenient form, e.g., a bar, a homogenous tablet, a tablet having two or more layers, a pouch having one or more compartments, a regular or compact powder, a granule, a paste, a gel, or a regular, compact or concentrated liquid. 
     Pouches can be configured as single or multicompartments. It can be of any form, shape and material which is suitable for hold the composition, e.g. without allowing the release of the composition to release of the composition from the pouch prior to water contact. The pouch is made from water soluble film which encloses an inner volume. Said inner volume can be divided into compartments of the pouch. Preferred films are polymeric materials preferably polymers which are formed into a film or sheet. Preferred polymers, copolymers or derivates thereof are selected polyacrylates, and water soluble acrylate copolymers, methyl cellulose, carboxy methyl cellulose, sodium dextrin, ethyl cellulose, hydroxyethyl cellulose, hydroxypropyl methyl cellulose, malto dextrin, poly methacrylates, most preferably polyvinyl alcohol copolymers and, hydroxypropyl methyl cellulose (HPMC). Preferably the level of polymer in the film for example PVA is at least about 60%. Preferred average molecular weight will typically be about 20,000 to about 150,000. Films can also be of blended compositions comprising hydrolytically degradable and water soluble polymer blends such as polylactide and polyvinyl alcohol (known under the Trade reference M8630 as sold by MonoSol LLC, Indiana, USA) plus plasticisers like glycerol, ethylene glycerol, propylene glycol, sorbitol and mixtures thereof. The pouches can comprise a solid laundry cleaning composition or part components and/or a liquid cleaning composition or part components separated by the water soluble film. The compartment for liquid components can be different in composition than compartments containing solids: US2009/0011970 A1. 
     Detergent ingredients can be separated physically from each other by compartments in water dissolvable pouches or in different layers of tablets. Thereby negative storage interaction between components can be avoided. Different dissolution profiles of each of the compartments can also give rise to delayed dissolution of selected components in the wash solution. 
     A liquid or gel detergent, which is not unit dosed, may be aqueous, typically containing at least 20% by weight and up to 95% water, such as up to about 70% water, up to about 65% water, up to about 55% water, up to about 45% water, up to about 35% water. Other types of liquids, including without limitation, alkanols, amines, diols, ethers and polyols may be included in an aqueous liquid or gel. An aqueous liquid or gel detergent may contain from 0-30% organic solvent. 
     The invention is further summarized in the following paragraphs:
         1. Use of at least one enzyme for preventing, reducing or removing odor from hard surface, wherein the at least one enzyme is selected from an alkaline cellulase and/or an enzyme having mannanase activity.   2. Use according to paragraph 1, wherein the alkaline cellulase is an enzyme exhibiting endo-beta-1,4-glucanase activity (EC 3.2.1.4)   3. Use according to paragraph 2, wherein the enzyme exhibiting endo-beta-1,4-glucanase activity has at least 80% sequence identity to SEQ ID NO: 1.   4. Use according to any of the preceding paragraphs, wherein the enzyme having mannanase activity has at least 80% sequence identity to SEQ ID NO: 2.   5. Use according to any of the preceding paragraphs, wherein the at least one enzyme is used together with an oxidase.   6. Use according to paragraph 5, wherein the oxidase is a carbohydrate oxidase (EC 1.1.3).   7. Use according to any of paragraphs 5-6, wherein the carbohydrate oxidase is a glucose oxidase (EC 1.1.3.4), a dehydrogenase a cellobiose oxidase (EC 1.1.99.18).   8. Use according to paragraph 7, wherein the glucose oxidase has at least 80% sequence identity to SEQ ID NO 3.   9. Use according to paragraph 7, wherein the cellobiose oxidase has at least 80% sequence identity to SEQ ID NO: 4.   10. Use according to any of the preceding paragraphs, wherein the odor is reduced by at least 50% when measured with Assay I and/or wherein the odor is scored on average 3 points lower than control when measured with Assay II.   11. Use according to paragraph 10, wherein the odor is reduced by at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% when measured with Assay I.   12. Use according to paragraph 10, wherein the odor is scored on average 4 points, 5 points, 6 points, 7 points or 8 points lower than control when measured with Assay II.   13. Use according to any of the preceding paragraphs, wherein the hard surface is a dish ware or a hard surface.   14. Use according to paragraph 13, wherein the hard surface is present in the interior of a dishwashing machine or a sink, such as walls, baskets, nozzles, pumps, sump, filters, pipelines, drains, and outlets.   15. Use according to any of paragraphs 1-14, wherein the enzymes are used in process for automatic dish wash or a process for manual dish wash.   16. A cleaning composition comprising a builder and at least one enzyme for preventing, reducing or removing odor from a hard surface, wherein the at least one enzyme is selected from an alkaline cellulase and/or an enzyme having mannanase activity.   17. Composition according to paragraph 16, wherein the composition is a dish wash composition.   18. Composition according to any of paragraphs 16 or 17, wherein the alkaline cellulase is an enzyme exhibiting endo-beta-1,4-glucanase activity (EC 3.2.1.4)   19. Composition according to paragraph 18, wherein the enzyme exhibiting endo-beta-1,4-glucanase activity has at least 80% sequence identity to SEQ ID NO: 1.   20. Composition according to any of paragraphs 16-19, wherein the enzyme having mannanase activity has at least 80% sequence identity to SEQ ID NO: 2.   21. Composition according to any of paragraphs 16-20, wherein the at least one enzyme is used together with an oxidase.   22. Composition according to paragraph 21, wherein the oxidase is a carbohydrate oxidase (EC 1.1.3).   23. Composition according to any of paragraphs 21-22, wherein the carbohydrate oxidase is a glucose oxidase (EC 1.1.3.4), a dehydrogenase or a cellobiose oxidase (EC 1.1.99.18).   24. Composition according to paragraph 23, wherein the glucose oxidase has at least 80% sequence identity to SEQ ID NO 4.   25. Composition according to paragraph 23, wherein the cellobiose oxidase has at least 80% sequence identity to SEQ ID NO: 4.   26. Composition according to any of paragraphs 16-25, wherein the composition comprises one or more enzymes in addition to the at least one enzymes and the oxidase.   27. Composition according to paragraph 26, wherein the composition comprises amylase and/or protease.   28. Composition according to paragraph 27, wherein the amylase is an alpha-amylase or a glucoamylase.   29. Composition according to paragraph 28, wherein the amylase has at least 80% sequence identity to SEQ ID NO: 5.   30. Composition according to paragraph 27, wherein the protease is a serine protease or a metalloprotease, preferably an alkaline microbial protease or a trypsin-like protease.   31. Composition according to paragraph 30, wherein the protease has at least 80% sequence identity to SEQ ID NO: 6 or the protease has at least 80% sequence identity to SEQ ID NO: 7.   32. Composition according to any of the preceding composition paragraphs, wherein the composition further comprises a surfactant.   33. Composition according to any of the preceding composition paragraphs, wherein the composition further comprises one or more builders and one or more polymer.   34. Composition according to any of the preceding composition composition paragraphs, wherein the composition further comprises one or more components selected from the group consisting of polymers, bleaching components, silicates, dyestuff and metal care agents.   35. Composition according to any of paragraphs 33-34, wherein the composition is a dishwashing composition in the form of a powder, a bar, a homogenous tablet, a tablet having two or more layers, a pouch having one or more compartments, a regular or compact powder, a granule, a paste, a gel, or a regular, compact or concentrated liquid.   36. Composition according to any of paragraphs 33-35, wherein the builder wherein the builder is selected among phosphates, sodium citrate builders, sodium carbonate, sodium silicate, sodium and zeolites.   37. Composition according to any of paragraphs 33-36, wherein the builder is added in an amount of about 0-65% by weight, preferably about 40-65% by weight, particularly about 20-65% by weight, particularly from 10% to 50% by weight.   38. Composition according to any of the preceding paragraphs, wherein the composition is phosphate free.   39. Composition according to any of paragraphs 33-38, wherein the builder is selected from citric acid, methyl glycine-N,N-diacetic acid (MGDA) and/or glutamic-N,N-diacetic acid (GLDA) and mixtures thereof.   40. Composition according to any of paragraphs 34-39, wherein wherein the bleach component is selected from bleaching catalysts, photobleaches, bleach activators, sources of hydrogen peroxide such as sodium percarbonate, sodium perborates and hydrogen peroxide, preformed peracids and mixtures thereof.   41. A method for washing an hard surface, wherein the method comprises the steps of:
           (i) Exposing the hard surface to a wash liquor comprising at least one enzyme selected from an alkaline cellulase and/or an enzyme having mannanase activity or the cleaning composition according to any of paragraphs 16-40, and   (ii) Rinsing the hard surface with water optionally comprising a rinsing aid;   wherein the hard surface is a dishware or a hard surface.   
           42. Method according to any of the preceding method paragraphs, wherein the hard surface is a hard surface in the interior of a dishwashing machine or a sink, such as walls, baskets, nozzles, pumps, sump, filters, pipelines, drains, and outlets.   43. Method according to any of paragraphs 41-42, wherein the method is hand dishwashing or automatic dish washing.   44. Method according to any of paragraphs 41-43, wherein the method is for washing or cleaning of dishware.   45. Method according to any of paragraphs 41-44, wherein the alkaline cellulase is an enzyme exhibiting endo-beta-1,4-glucanase activity (EC 3.2.1.4)   46. Method according to paragraph 45, wherein the enzyme exhibiting endo-beta-1,4-glucanase activity has at least 80% sequence identity to SEQ ID NO: 1.   47. Method according to any of the preceding method paragraphs, wherein the enzyme having mannanase activity has at least 80% sequence identity to SEQ ID NO: 2.   48. Method according to any of the preceding method paragraphs, wherein the at least one enzyme is used together with an oxidase.   49. Method according to paragraph 48, wherein the oxidase is a carbohydrate oxidase (EC 1.1.3).   50. Method according to any of paragraphs 48-49, wherein the carbohydrate oxidase is a glucose oxidase (EC 1.1.3.4), a dehydrogenase, a cellobiose oxidase (EC 1.1.99.18)   51. Method according to paragraph 50, wherein the glucose oxidase has at least 80% sequence identity to SEQ ID NO: 3.   52. Method according to paragraph 50, wherein the cellobiose oxidase has at least 80% sequence identity to SEQ ID NO: 4.   53. Method according to any of paragraphs 41-52, wherein the composition comprises one or more enzymes in addition to the at least one enzymes and the oxidase.   54. Method according to paragraph 53, wherein the composition comprises amylase and/or protease.   55. Method according to paragraph 54, wherein the amylase is an alpha-amylase or a glucoamylase.   56. Method according to any of paragraphs 51-55, wherein the amylase has at least 80% sequence identity to SEQ ID NO: 5.   57. Method according to paragraph 54, wherein the protease is chemically modified or protein engineered.   58. Method according to paragraph any of paragraphs 54 and 57, wherein the protease is a serine protease or a metalloprotease, preferably an alkaline microbial protease or a trypsin-like protease.   59. Method according to paragraph 58, wherein the protease has at least 80% sequence identity to SEQ ID NO: 6 or the protease has at least 80% sequence identity to SEQ ID NO: 7.   60. Method according to any of the preceding method paragraphs, wherein the composition further comprises a surfactant.   61. Method according to any of the preceding method paragraphs, wherein the composition further comprises one or more builders and one or more polymer.   62. Method according to any of the preceding method composition paragraphs, wherein the composition further comprises one or more components selected from the group consisting of polymers, bleaching systems, bleach activators, bleach catalysts, silicates, dyestuff and metal care agents.   63. Method according to any of the preceding method paragraphs, wherein the method is an automatic dishwashing method or a method for manual dish washing.       

     Detergent Compositions 
     The present ADW detergent compositions can be used together with the enzymes of the invention. 
     SUN prof dishwash tablets P K100 HG756: 5-45% oxygen based bleaching agents and less than 5% non-ionic surfactant, phosphonates and polycarboxylates. 
     SUN—Sunlight Auto Dish Tablet Detergent—Lemon Oxi Work in Progress: Sodium Carbonate Sodium Cholride, Sodium Citrate, Sodium Carbonate Peroxide, Sodium Polycarboxylate, Sodium Silicate, Alcohol Alkoxylate, Enzyme, Water, Perfume, Benzotriazole and Liquiting® Blue HP. 
     SUN—Sunlight Auto Dish Powder Detergent—Lemon Oxi: Sodium Cholride, Sodium Carbonate, Sodium Silicate, Sodium Polycarboxylate, Sodium Gluconate, Sodium Carbonate peroxide, Alcohol Alkoxylate, Sodium Silicoaluminate, Enzyme, Perfume, Water, Liquitint Yellow and Liquiting® Blue HP. 
     SUN—Auto Dish Gel Detergent—citrus: Water, Sodium Silicate, Sodium Hypochlorite, Sodium Carbonate, Sodium Polycarboxylate, Sodium Hydroxide, Carbomer, Sodium benzoate, Sulfuric Acid, Fragance. 
     SUN—Sunlight Ultra Liquid Dish Detergent—Green Apple: Sodium Dedecylbenzenesulfonate, Sodium Laureth Sulfate, Cocamidopropyl Betaine, Sodium Hydroxide, Magnesium Sulfate, Sodium Xylenesulfonate, Alcohol Denatured, Perfume, Methylchloroisothiazolinone, Methylisothiazolinone, Sanolin® Yellow E-2GL and Sanolin® Blue NBL. 
     Cascade Platinum® Action Pacs®: Powder ingredients: Sodium Carbonate, Sodium Percarbonate, Sodium Silicate, Modified Plyacrylate, Methyl Glycine deacetic Acid (trisodium Salt), Sodium Sulfate, Protease Enzyme, Amylase Enzyme, Alcohol Alkoxylate, Plyethylene glycol, Hydrozincite, Amine Cobalt Salt, Water, Perfumes. Liquid ingredients: Alcohol Alkoxylate, Trideceth-n, Dipropylene glycol, water, glycerine, Acid Red #33 and/or FD&amp;C Yellow #5 and/or Acid Blue 182 and/or Dye Reactive Green 12. Film: Polyvinyl Alcohol Copolymer. 
     Cascade Gel: Water, Sodium Silicate, Sodium Carbonate, Sodium Sulfonate, Sodium Polyacrylate, Cross-linked polyacrylate, Sodium Benzoate, Sodium Hypocholrite, Zinc Carbonate, Sodium Hydroxide, Nitric acid, Perfume. 
     Cascade Powder: Sodium Carbonate, Sodium Sulfate, Polyethyleneimine (sulfonated), Sodium 
     Silicate, Water, Alcohol Alkoxylate, Sodium Percarbonate, Protease Enzyme, Amylase Enzyme, Amine Cobalt Salt and Perfumes. 
     Assays 
     Assay I 
     Analysis of Gas Composition Using E-Nose (GC Headspace). 
     As part of the testing real food items were incubated overnight after being prepared as outlined under examples. The headspace was allowed to accumulate in gas vials and the headspace from the 2 mL analyzed from the capped vials in a Heracles II Flash Gas Chromatography Electronic nose from Alpha M.O.S., France (double column gas chromatograph with 2 FIDs, column 1: MXT5 and column 2: MXT1701) after 20 minutes incubation at 40° C. 
     Assay II 
     Odor Removal (Sensory Analysis) 
     After being prepared as outlined under examples and incubated overnight, the vials were then allowed to develop malodor over an additional 2 days to allow for an odor panel to evaluate. The odor panel consists of trained panelists, who were presented with a blind evaluation. T. The odor panel this time was asked to rank the odor from 1 to 5, where 5 was the most pungent odor, and 1 was not significantly different from the blank odor. 
     Examples 
     Preparation of Foodstuffs for Odor Generation 
     The following foods were identified to be likely causes of malodor in automatic dishwashing machines. Broccoli, Egg, Fish, Onion, and Garlic. Foods were purchased from a local supermarket (Netto, Bagsværd) and initially prepared in the following manner. 
     Fresh Broccoli: 2 florets of broccoli were covered in water and boiled for 10 min. The broccoli was rinsed in cold water and left to cool. The 2 florets were blended to homogeneity using a hand blender and collected in a petri dish for processing. 
     Eggs: 1-2 eggs were fried is hot sun flower oil for 1.5 min each side to the appearance of “sunny side up” (yolk slightly runny, not a hard yolk). 
     Garlic and Onion were untreated and used raw after peeling. 
     The Fish was purchased as a processed Fishball (Bådsmand Fiskedeller) and one half of one ball processed to homogeneity in the hand blender. 
     Processing of Foodstuff after Initial Preparation. 
     Assay Buffer: Buffer pH 8.00. 50 mM HEPES, 50 mM CHES, 1 mM CaCl 2 ), 0.01% triton X-100. 
     Water hardness at 21° dH was added to the buffer. The Ca:NaHCO 3 :Mg ratio was 4:2:1, prepared using 4000° dH CaCl 2 ), 2000° dH MgCl2 stock solutions and freshly prepared 0.535 M NaHCO 3 . 
     All the initially prepared food were treated as described below in order to obtain substrate to be tested:
         5 g of blended food as prepared above was added to 100 mL of assay buffer and mixed to a fine liquid suspension (ca. 3 min of blending using a handheld 600 watt turbo Braun blender).   8 replicates of 2 mL were pipetted to a 20 mL GC headspace vial. To 4 replicates the enzymes were added, and the other 4 served as a control.   Each foodsample was prepared individually first and after pipetting the lids screwed tightly in place.   The vials were mixed using a whirl mixer for 30 s at 2500 rpm.   Controls are listed below.   GC Vials were prepared with buffer alone and with buffer and enzymes only but without food to serve as background gas controls.   All vials were incubated at 37° C. over 24 h.       

                     TABLE 1                  Experimental setup                                     Label   Food   Enzymes   Comment                       A 1-4   None   None   Control           B 1-4   None   All   Control           C 1-4   None   All (SEQ 6)   Control           D 1-4   Egg   None   Control           E 1-4   Garlic   None   Control           F 1-4   Onion   None   Control           G 1-4   Fish   None   Control           H 1-4   Broccoli   None   Control           I 1-4   Egg   All (SEQ 6)           J 1-4   Garlic   ALL           K 1-4   Onion   ALL           L 1-4   Fish   ALL           M 1-4   Broccoli   ALL                        
None means that no enzymes are added. All means that enzymes SEQ ID NO: 1, 2, 4, 5 and 7 were added in the below concentrations. All (SEQ 6) means that enzymes SEQ ID NO: 1, 2, 4, 5 and 6 were added
 
     
       
         
           
               
               
               
             
               
                   
                 TABLE 2 
               
               
                   
                   
               
               
                   
                 Enzyme 
                 mg/L in assay 
               
               
                   
                   
               
             
            
               
                   
               
            
           
           
               
               
               
            
               
                   
                 SEQ ID NO: 1 
                 0.1 
               
               
                   
                 SEQ ID NO: 2 
                 30.8 
               
               
                   
                 SEQ ID NO: 4 
                 105.0 
               
               
                   
                 SEQ ID NO: 6 
                 6.4 
               
               
                   
                 SEQ ID NO: 7 
                 6.5 
               
               
                   
                 SEQ ID NO: 5 
                 36.4 
               
               
                   
                   
               
            
           
         
       
     
     Example 1 
     The experimental set up is shown tables 1-2 above. 
     Analysis of Gas Composition Using E-Nose (Assay I). 
     As part of the testing real food items were incubated overnight after being prepared as outlined above. The headspace was allowed to accumulate in gas vials and the headspace from the 2 mL analyzed from the capped vials in a Heracles II Flash Gas Chromatography Electronic nose from Alpha M.O.S., France (double column gas chromatograph with 2 FIDs, column 1: MXT5 and column 2: MXT1701) after 20 minutes incubation at 40° C. 
     Evaluation of Odor Removal 
     After overnight incubation, the foodstuffs were analyzed by E-Nose as described above; the vials were then allowed to develop malodor over an additional 2 d to allow for an odor panel to evaluate. The odor panel consist of 5 trained panelist, who were presented with a blind evaluation. The panelists were asked to evaluate the difference in malodor on a scale of N/A to +++, where N/A was not applicable due to over generation of scent in both control and enzyme treated vials, and the difference if detected could be ranked thusly: 
     
       
         
           
               
               
               
             
               
                   
                   
               
               
                   
                 Symbol 
                 Key 
               
               
                   
                   
               
             
            
               
                   
                 +++ 
                 Almost no residual gas/odour left 
               
               
                   
                 ++ 
                 Very slight gas/odour remaining 
               
               
                   
                 N/A 
                 Out of range 
               
               
                   
                   
               
            
           
         
       
     
     These results were compared to the E-Nose data where the integral of the areas was used to determine large differences. 
     
       
         
           
               
               
               
               
             
               
                   
                   
               
               
                   
                   
                 E-Nose: 
                 Panel: 
               
               
                   
                   
                 Gas 
                 Odor 
               
               
                   
                 Food 
                 Reduction 
                 Reduction 
               
               
                   
                   
               
             
            
               
                   
                 Broccoli 
                 +++ 
                 +++ 
               
               
                   
                 Fish 
                 +++ 
                 +++ 
               
               
                   
                 Onion 
                 N/A 
                 N/A 
               
               
                   
                 Egg 
                 ++ 
                 +++ 
               
               
                   
                 Garlic 
                 N/A 
                 N/A 
               
               
                   
                   
               
            
           
         
       
     
     Neither onion nor Garlic could be evaluated due to the signal being too high for both the E-nose and the panelists. 
     
       
         
           
               
             
               
                 TABLE 3 
               
               
                   
               
               
                 GC Headspace retention measurements for food items with and without enzymes. Channel 1, a cutoff value of  
               
               
                 10,000 was chosen to pick the most relevant peaks, similar trends were seen in channel 2 (data not shown). 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                 GC HeadSpace Retention 
                 11.87-1- 
                 12.03-1- 
                 12.16-1- 
                 12.74-1- 
                 13.89-1- 
                 14.03-1- 
                 15.07-1- 
                 16.99-1- 
               
               
                 Time-Channel 1 
                 A 
                 A 
                 A 
                 A 
                 A 
                 A 
                 A 
                 A 
               
               
                   
               
               
                 Buffer only Ctrl 
                 0 
                 0 
                 0 
                 3949 
                 0 
                 0 
                 628 
                 355 
               
               
                 All in buffer Ctrl 
                 0 
                 0 
                 0 
                 4229 
                 0 
                 0 
                 5346 
                 6653 
               
               
                 All (SEQ 6) in buffer Ctrl 
                 0 
                 0 
                 0 
                 4270 
                 0 
                 0 
                 3583 
                 5464 
               
               
                 Egg in buffer 
                 0 
                 0 
                 0 
                 4003 
                 363 
                 528 
                 3290 
                 17687 
               
               
                 Garlic in buffer 
                 1125611 
                 0 
                 1182473 
                 0 
                 0 
                 0 
                 0 
                 220742 
               
               
                 Onion in buffer 
                 0 
                 0 
                 0 
                 18071 
                 0 
                 2615 
                 44361 
                 213736 
               
               
                 Fish in buffer 
                 0 
                 0 
                 0 
                 8749 
                 0 
                 0 
                 41483 
                 7217 
               
               
                 Broccoli in buffer 
                 0 
                 0 
                 0 
                 3828 
                 22626 
                 15037 
                 0 
                 1464 
               
               
                 Egg with All (SEQ 6) 
                 0 
                 0 
                 0 
                 4095 
                 0 
                 0 
                 4312 
                 5941 
               
               
                 Garlic with All 
                 485915 
                 497499 
                 0 
                 916651 
                 0 
                 0 
                 15269 
                 192084 
               
               
                 Onion with All 
                 0 
                 0 
                 0 
                 16537 
                 0 
                 0 
                 6596 
                 229689 
               
               
                 Fish with All 
                 25 
                 0 
                 0 
                 6489 
                 0 
                 0 
                 5001 
                 28035 
               
               
                 Broccoli with All 
                 0 
                 0 
                 0 
                 9590 
                 0 
                 0 
                 5917 
                 10958 
               
               
                   
               
               
                 GC HeadSpace Retention 
                 17.52-1- 
                 18.57-1- 
                 18.94-1- 
                 21.17-1- 
                 22.27-1- 
                 26.88-1- 
                 28.45-1- 
                 30.06-1- 
               
               
                 Time-Channel 1 
                 A 
                 A 
                 A 
                 A 
                 A 
                 A 
                 A 
                 A 
               
               
                   
               
               
                 Buffer only Ctrl 
                 0 
                 0 
                 0 
                 10676 
                 0 
                 240 
                 0 
                 0 
               
               
                 All in buffer Ctrl 
                 0 
                 0 
                 0 
                 11466 
                 0 
                 732 
                 118 
                 0 
               
               
                 All (SEQ 6) in buffer Ctrl 
                 0 
                 0 
                 897 
                 8454 
                 0 
                 640 
                 0 
                 0 
               
               
                 Egg in buffer 
                 0 
                 0 
                 0 
                 10602 
                 0 
                 663 
                 54 
                 0 
               
               
                 Garlic in buffer 
                 0 
                 37560 
                 39058 
                 179301 
                 0 
                 452650 
                 0 
                 237232 
               
               
                 Onion in buffer 
                 0 
                 0 
                 0 
                 7810 
                 40585 
                 3221 
                 93060 
                 0 
               
               
                 Fish in buffer 
                 1073 
                 1892 
                 4037 
                 13502 
                 0 
                 1811 
                 1463 
                 0 
               
               
                 Broccoli in buffer 
                 0 
                 0 
                 94909 
                 16448 
                 0 
                 1687 
                 302 
                 0 
               
               
                 Egg with All (SEQ 6) 
                 0 
                 0 
                 1226 
                 11016 
                 0 
                 665 
                 102 
                 0 
               
               
                 Garlic with All 
                 0 
                 16191 
                 36397 
                 185402 
                 0 
                 398142 
                 0 
                 240635 
               
               
                 Onion with All 
                 77881 
                 0 
                 0 
                 12363 
                 4787 
                 2087 
                 10948 
                 0 
               
               
                 Fish with All 
                 0 
                 0 
                 0 
                 9100 
                 0 
                 1346 
                 0 
                 0 
               
               
                 Broccoli with All 
                 0 
                 0 
                 4124 
                 11274 
                 0 
                 5198 
                 164 
                 0 
               
               
                   
               
               
                 GC HeadSpace Retention 
                 34.06-1- 
                 36.40-1- 
                 38.18-1- 
                 44.88-1- 
                 54.38-1- 
                 56.63-1- 
                 69.88-1- 
                 77.50-1- 
               
               
                 Time-Channel 1 
                 A 
                 A 
                 A 
                 A 
                 A 
                 A 
                 A 
                 A 
               
               
                   
               
               
                 Buffer only Ctrl 
                 0 
                 0 
                 0 
                 1551 
                 0 
                 251 
                 244 
                 0 
               
               
                 All in buffer Ctrl 
                 0 
                 173 
                 134 
                 1522 
                 0 
                 1346 
                 145 
                 0 
               
               
                 All (SEQ 6) in buffer Ctrl 
                 0 
                 110 
                 110 
                 1498 
                 0 
                 1260 
                 172 
                 0 
               
               
                 Egg in buffer 
                 0 
                 1599 
                 1276 
                 3001 
                 8547 
                 2161 
                 6351 
                 0 
               
               
                 Garlic in buffer 
                 62324 
                 0 
                 37158 
                 252484 
                 1126 
                 35733 
                 21430 
                 824 
               
               
                 Onion in buffer 
                 99 
                 0 
                 37 
                 3954 
                 92 
                 117007 
                 1074 
                 17971 
               
               
                 Fish in buffer 
                 0 
                 3331 
                 1293 
                 1723 
                 22259 
                 3349 
                 602 
                 113 
               
               
                 Broccoli in buffer 
                 0 
                 0 
                 409 
                 4225 
                 34 
                 298734 
                 860 
                 0 
               
               
                 Egg with All (SEQ 6) 
                 0 
                 1017 
                 531 
                 2095 
                 22 
                 5365 
                 1057 
                 0 
               
               
                 Garlic with All 
                 30806 
                 15894 
                 38259 
                 232711 
                 1747 
                 46476 
                 60232 
                 804 
               
               
                 Onion with All 
                 50 
                 590 
                 515 
                 1953 
                 0 
                 16481 
                 1101 
                 21868 
               
               
                 Fish with All 
                 0 
                 1571 
                 1700 
                 2173 
                 36 
                 7481 
                 2169 
                 780 
               
               
                 Broccoli with All 
                 0 
                 634 
                 300 
                 31935 
                 0 
                 3640 
                 1091 
                 0 
               
               
                   
               
               
                 GC HeadSpace Retention 
                 83.31-1- 
                 85.96-1- 
                 88.63-1- 
                 90.71-1- 
                 92.36-1- 
                 94.70-1- 
                 97.18-1- 
                 99.10-1- 
               
               
                 Time-Channel 1 
                 A 
                 A 
                 A 
                 A 
                 A 
                 A 
                 A 
                 A 
               
               
                   
               
               
                 Buffer only Ctrl 
                 0 
                 0 
                 203 
                 1734 
                 0 
                 0 
                 240 
                 176 
               
               
                 All in buffer Ctrl 
                 16 
                 18 
                 228 
                 1542 
                 0 
                 0 
                 520 
                 182 
               
               
                 All (SEQ 6) in buffer Ctrl 
                 13 
                 65 
                 220 
                 1551 
                 0 
                 0 
                 307 
                 154 
               
               
                 Egg in buffer 
                 0 
                 2320 
                 493 
                 2325 
                 0 
                 0 
                 125 
                 164 
               
               
                 Garlic in buffer 
                 1744985 
                 19533 
                 136277 
                 75352 
                 77217 
                 1151297 
                 32141 
                 204010 
               
               
                 Onion in buffer 
                 791 
                 3505 
                 199 
                 1418 
                 2021 
                 11674 
                 345581 
                 22052 
               
               
                 Fish in buffer 
                 237 
                 227 
                 482 
                 1331 
                 103 
                 51 
                 546 
                 97 
               
               
                 Broccoli in buffer 
                 139 
                 0 
                 215 
                 1585 
                 0 
                 912 
                 206 
                 624 
               
               
                 Egg with All (SEQ 6) 
                 0 
                 117 
                 118 
                 2138 
                 0 
                 0 
                 190 
                 135 
               
               
                 Garlic with All 
                 1915336 
                 17607 
                 166169 
                 87537 
                 130011 
                 915011 
                 40447 
                 273854 
               
               
                 Onion with All 
                 1044 
                 1191 
                 266 
                 2091 
                 3027 
                 3086 
                 88181 
                 31242 
               
               
                 Fish with All 
                 264 
                 229 
                 178 
                 2904 
                 144 
                 47 
                 602 
                 128 
               
               
                 Broccoli with All 
                 176 
                 70 
                 121 
                 2177 
                 0 
                 118 
                 174 
                 155 
               
               
                   
               
            
           
           
               
               
               
               
               
               
            
               
                 GC HeadSpace Retention 
                 102.86- 
                 105.84- 
                 129.34- 
                 159.06- 
                 188.44- 
               
               
                 Time-Channel 1 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
               
               
                   
               
               
                 Buffer only Ctrl 
                 340 
                 2514 
                 5738 
                 13325 
                 3686 
               
               
                 All in buffer Ctrl 
                 386 
                 5563 
                 5062 
                 18778 
                 2171 
               
               
                 All (SEQ 6) in buffer Ctrl 
                 365 
                 3207 
                 5170 
                 15306 
                 1799 
               
               
                 Egg in buffer 
                 349 
                 3624 
                 4710 
                 9146 
                 1582 
               
               
                 Garlic in buffer 
                 936179 
                 0 
                 2498891 
                 3516955 
                 234935 
               
               
                 Onion in buffer 
                 2905 
                 35732 
                 488812 
                 214933 
                 25010 
               
               
                 Fish in buffer 
                 576 
                 6296 
                 9820 
                 19769 
                 2069 
               
               
                 Broccoli in buffer 
                 491 
                 5367 
                 6442 
                 10633 
                 1502 
               
               
                 Egg with All (SEQ 6) 
                 152 
                 1989 
                 4243 
                 15925 
                 1216 
               
               
                 Garlic with All 
                 702074 
                 21502 
                 4110087 
                 3378480 
                 258596 
               
               
                 Onion with All 
                 2625 
                 10597 
                 233400 
                 102226 
                 14717 
               
               
                 Fish with All 
                 837 
                 4473 
                 11156 
                 28055 
                 1980 
               
               
                 Broccoli with All 
                 489 
                 2792 
                 6568 
                 18034 
                 1423 
               
               
                   
               
            
           
         
       
     
     Example 2 
     Example 1 was repeated with the following differences: the Onion and Garlic were processed at 25 mg/2 mL. The experimental set up is shown tables 1-2 above. 
     Data from Assay I can be found in table 4 below. 
     
       
         
           
               
             
               
                 TABLE 4 
               
               
                   
               
               
                 Assay I: GC Headspace retention measurements for food items with and without enzymes. Channel 1, a cutoff value  
               
               
                 of 10,000 was chosen to pick the most relevant peaks, similar trends were seen in channel 2 (data not shown). 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                 GC HeadSpace Retention 
                 12.33-1- 
                 12.45-1- 
                 12.60-1- 
                 14.43-1- 
                 15.84-1- 
                 17.32-1- 
                 17.44-1- 
                 18.46-1- 
               
               
                 Time-Channel 1 
                 A 
                 A 
                 A 
                 A 
                 A 
                 A 
                 A 
                 A 
               
               
                   
               
               
                 Buffer only Ctrl 
                 0 
                 0 
                 0 
                 0 
                 368 
                 0 
                 0 
                 13758 
               
               
                 All in buffer Ctrl 
                 0 
                 0 
                 0 
                 2994 
                 9263 
                 0 
                 0 
                 15193 
               
               
                 All (SEQ 6) in buffer Ctrl 
                 0 
                 0 
                 0 
                 3074 
                 9215 
                 0 
                 0 
                 15515 
               
               
                 Egg in buffer 
                 0 
                 0 
                 0 
                 5178 
                 2283 
                 0 
                 0 
                 15147 
               
               
                 Garlic in buffer 
                 114467 
                 317782 
                 0 
                 0 
                 67904 
                 0 
                 0 
                 40664 
               
               
                 Onion in buffer 
                 0 
                 0 
                 0 
                 0 
                 56685 
                 0 
                 0 
                 10686 
               
               
                 Fish in buffer 
                 0 
                 0 
                 0 
                 366929 
                 0 
                 0 
                 0 
                 12137 
               
               
                 Broccoli in buffer 
                 0 
                 0 
                 0 
                 32100 
                 0 
                 78289 
                 26344 
                 0 
               
               
                 Egg with All (SEQ 6) 
                 0 
                 0 
                 0 
                 3507 
                 10315 
                 0 
                 0 
                 13053 
               
               
                 Garlic with All 
                 0 
                 165671 
                 54238 
                 0 
                 59851 
                 0 
                 0 
                 33880 
               
               
                 Onion with All 
                 0 
                 0 
                 0 
                 3487 
                 105256 
                 0 
                 0 
                 23924 
               
               
                 Fish with All 
                 0 
                 0 
                 0 
                 5549 
                 32913 
                 0 
                 0 
                 13997 
               
               
                 Broccoli with All 
                 0 
                 0 
                 0 
                 4571 
                 15139 
                 0 
                 0 
                 15743 
               
               
                   
               
               
                 GC HeadSpace Retention 
                 18.90-1- 
                 19.39-1- 
                 20.45-1- 
                 22.77-1- 
                 24.45-1- 
                 25.51-1- 
                 37.35-1- 
                 44.94-1- 
               
               
                 Time-Channel 1 
                 A 
                 A 
                 A 
                 A 
                 A 
                 A 
                 A 
                 A 
               
               
                   
               
               
                 Buffer only Ctrl 
                 0 
                 0 
                 0 
                 700 
                 0 
                 0 
                 0 
                 0 
               
               
                 All in buffer Ctrl 
                 0 
                 0 
                 0 
                 1103 
                 0 
                 0 
                 0 
                 0 
               
               
                 All (SEQ 6) in buffer Ctrl 
                 0 
                 0 
                 0 
                 1234 
                 0 
                 0 
                 0 
                 0 
               
               
                 Egg in buffer 
                 0 
                 0 
                 0 
                 1149 
                 0 
                 0 
                 1227 
                 1681 
               
               
                 Garlic in buffer 
                 0 
                 0 
                 0 
                 96821 
                 0 
                 40173 
                 37774 
                 0 
               
               
                 Onion in buffer 
                 0 
                 20194 
                 0 
                 1458 
                 27290 
                 0 
                 0 
                 0 
               
               
                 Fish in buffer 
                 0 
                 21596 
                 0 
                 4020 
                 7163 
                 0 
                 8754 
                 10496 
               
               
                 Broccoli in buffer 
                 0 
                 0 
                 0 
                 803 
                 0 
                 0 
                 6778 
                 1764 
               
               
                 Egg with All (SEQ 6) 
                 0 
                 0 
                 0 
                 866 
                 0 
                 0 
                 0 
                 0 
               
               
                 Garlic with All 
                 12027 
                 0 
                 0 
                 62820 
                 0 
                 24526 
                 30472 
                 0 
               
               
                 Onion with All 
                 0 
                 0 
                 0 
                 1588 
                 6767 
                 0 
                 85 
                 0 
               
               
                 Fish with All 
                 0 
                 0 
                 15346 
                 0 
                 0 
                 0 
                 3348 
                 0 
               
               
                 Broccoli with All 
                 0 
                 0 
                 0 
                 8136 
                 0 
                 0 
                 25140 
                 0 
               
               
                   
               
               
                 GC HeadSpace Retention 
                 47.47-1- 
                 58.24-1- 
                 70.53-1- 
                 75.42-1- 
                 77.24-1- 
                 79.42-1- 
                 81.22-1- 
                 84.03-1- 
               
               
                 Time-Channel 1 
                 A 
                 A 
                 A 
                 A 
                 A 
                 A 
                 A 
                 A 
               
               
                   
               
               
                 Buffer only Ctrl 
                 0 
                 184 
                 0 
                 261 
                 1743 
                 15 
                 0 
                 53 
               
               
                 All in buffer Ctrl 
                 0 
                 152 
                 0 
                 140 
                 1156 
                 15 
                 0 
                 126 
               
               
                 All (SEQ 6) in buffer Ctrl 
                 0 
                 148 
                 0 
                 270 
                 1674 
                 0 
                 0 
                 132 
               
               
                 Egg in buffer 
                 247 
                 2513 
                 0 
                 254 
                 1904 
                 0 
                 0 
                 104 
               
               
                 Garlic in buffer 
                 11946 
                 6596 
                 324772 
                 17996 
                 18052 
                 10348 
                 190882 
                 7091 
               
               
                 Onion in buffer 
                 3092 
                 223 
                 255 
                 179 
                 1219 
                 379 
                 1562 
                 19836 
               
               
                 Fish in buffer 
                 0 
                 5971 
                 82 
                 280 
                 1052 
                 31 
                 55 
                 1278 
               
               
                 Broccoli in buffer 
                 30284 
                 2180 
                 0 
                 686 
                 1648 
                 16 
                 95 
                 317 
               
               
                 Egg with All (SEQ 6) 
                 0 
                 1865 
                 0 
                 177 
                 2063 
                 31 
                 0 
                 172 
               
               
                 Garlic with All 
                 11183 
                 11158 
                 252367 
                 11225 
                 12751 
                 7712 
                 120800 
                 4657 
               
               
                 Onion with All 
                 0 
                 1379 
                 211 
                 249 
                 1549 
                 359 
                 189 
                 2876 
               
               
                 Fish with All 
                 0 
                 5678 
                 81 
                 333 
                 3109 
                 38 
                 0 
                 739 
               
               
                 Broccoli with All 
                 0 
                 1720 
                 15 
                 214 
                 2206 
                 0 
                 0 
                 203 
               
               
                   
               
               
                 GC HeadSpace Retention 
                 85.37-1- 
                 89.06-1- 
                 109.13- 
                 114.62- 
                 118.39- 
                 123.45- 
                 129.00- 
                 137.22- 
               
               
                 Time-Channel 1 
                 A 
                 A 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
               
               
                   
               
               
                 Buffer only Ctrl 
                 323 
                 556 
                 5852 
                 1790 
                 1019 
                 710 
                 3654 
                 1076 
               
               
                 All in buffer Ctrl 
                 312 
                 623 
                 5516 
                 1630 
                 841 
                 228 
                 10280 
                 919 
               
               
                 All (SEQ 6) in buffer Ctrl 
                 314 
                 695 
                 5769 
                 1620 
                 1255 
                 180 
                 11184 
                 761 
               
               
                 Egg in buffer 
                 316 
                 512 
                 6289 
                 1555 
                 569 
                 233 
                 1918 
                 712 
               
               
                 Garlic in buffer 
                 33171 
                 193008 
                 3130042 
                 76838 
                 36723 
                 72911 
                 75586 
                 159679 
               
               
                 Onion in buffer 
                 3942 
                 808 
                 64165 
                 20089 
                 38742 
                 585 
                 3101 
                 1183 
               
               
                 Fish in buffer 
                 250 
                 863 
                 7392 
                 2012 
                 426 
                 190 
                 1854 
                 956 
               
               
                 Broccoli in buffer 
                 482 
                 635 
                 5308 
                 1545 
                 433 
                 177 
                 2527 
                 815 
               
               
                 Egg with All (SEQ 6) 
                 317 
                 677 
                 6009 
                 1718 
                 630 
                 323 
                 13123 
                 653 
               
               
                 Garlic with All 
                 25049 
                 102954 
                 2978852 
                 45903 
                 17777 
                 35537 
                 58083 
                 96108 
               
               
                 Onion with All 
                 1174 
                 877 
                 23147 
                 7519 
                 1989 
                 572 
                 12976 
                 1753 
               
               
                 Fish with All 
                 293 
                 896 
                 12225 
                 3231 
                 472 
                 611 
                 13520 
                 2790 
               
               
                 Broccoli with All 
                 295 
                 964 
                 5648 
                 2380 
                 377 
                 471 
                 12934 
                 1211 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
            
               
                 GC HeadSpace Retention 
                 143.24- 
                 154.28- 
                 169.95- 
                 191.97- 
                 215.61- 
                 218.92- 
                 229.13- 
               
               
                 Time-Channel 1 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
               
               
                   
               
               
                 Buffer only Ctrl 
                 1717 
                 4478 
                 1819 
                 8656 
                 0 
                 0 
                 0 
               
               
                 All in buffer Ctrl 
                 1747 
                 5068 
                 1845 
                 7366 
                 0 
                 0 
                 0 
               
               
                 All (SEQ 6) in buffer Ctrl 
                 1761 
                 5122 
                 1959 
                 8306 
                 0 
                 0 
                 0 
               
               
                 Egg in buffer 
                 1140 
                 4204 
                 704 
                 7062 
                 0 
                 0 
                 0 
               
               
                 Garlic in buffer 
                 71647 
                 88584 
                 27466 
                 23567 
                 1855825 
                 0 
                 908875 
               
               
                 Onion in buffer 
                 75943 
                 5797 
                 4409 
                 10753 
                 527175 
                 0 
                 2612 
               
               
                 Fish in buffer 
                 2052 
                 9444 
                 709 
                 6980 
                 0 
                 0 
                 661775 
               
               
                 Broccoli in buffer 
                 1593 
                 5464 
                 1010 
                 7032 
                 13406 
                 0 
                 2672 
               
               
                 Egg with All (SEQ 6) 
                 1019 
                 3929 
                 816 
                 5495 
                 0 
                 0 
                 3385 
               
               
                 Garlic with All 
                 47660 
                 52839 
                 17170 
                 20449 
                 3769 
                 0 
                 1711225 
               
               
                 Onion with All 
                 12650 
                 6056 
                 2121 
                 9793 
                 620925 
                 228725 
                 2826 
               
               
                 Fish with All 
                 2179 
                 9386 
                 734 
                 7549 
                 0 
                 0 
                 260025 
               
               
                 Broccoli with All 
                 1162 
                 4835 
                 656 
                 7448 
                 1097125 
                 0 
                 0 
               
               
                   
               
            
           
         
       
     
     Example 3 
     In addition to example 2, a parallel trial was set up to determine which enzymes performed the odor reduction most effectively. 
     Broccoli, fish and egg as well as a background buffer blank was set up in the following manner 
     
       
         
           
               
             
               
                 TABLE 5 
               
             
            
               
                   
               
               
                 showing Individual enzyme analysis 
               
            
           
           
               
               
               
               
               
               
            
               
                   
                   
                   
                 Manna- 
                 Cellu- 
                   
               
               
                   
                   
                   
                 nase 
                 lase 
                 Oxidase 
               
               
                   
                   
                   
                 SEQ ID 
                 SEQ ID 
                 SEQ ID 
               
               
                 Label 
                 Food 
                 Enzymes 
                 NO: 2 
                 NO: 1 
                 NO: 4 
               
               
                   
               
               
                 XX A 1-4 
                 Differs 
                 None 
                   
                   
                   
               
               
                 XX B 1-4 
                 Differs 
                 Protease (SEQ ID 
               
               
                   
                   
                 NO: 6 or 7) + 
               
               
                   
                   
                 Amylase (SEQ ID 
               
               
                   
                   
                 NO: 5) 
               
               
                 XX C 1-4 
                 Differs 
                 Protease (SEQ ID 
                 X 
               
               
                   
                   
                 NO: 6 or 7) + 
               
               
                   
                   
                 Amylase (SEQ ID 
               
               
                   
                   
                 NO: 5) 
               
               
                 XX D 1-4 
                 Differs 
                 Protease (SEQ ID 
                   
                 X 
               
               
                   
                   
                 NO: 6 or 7) + 
               
               
                   
                   
                 Amylase (SEQ ID 
               
               
                   
                   
                 NO: 5) 
               
               
                 XX E 1-4 
                 Differs 
                 Protease (SEQ ID 
                   
                   
                 X 
               
               
                   
                   
                 NO: 6 or 7) + 
               
               
                   
                   
                 Amylase (SEQ ID 
               
               
                   
                   
                 NO: 5) 
               
               
                 XX F 1-4 
                 Differs 
                 Protease (SEQ ID 
                 X 
                 X 
               
               
                   
                   
                 NO: 6 or 7) + 
               
               
                   
                   
                 Amylase (SEQ ID 
               
               
                   
                   
                 NO: 5) 
               
               
                 XX G 1-4 
                 Differs 
                 Protease (SEQ ID 
                   
                 X 
                 X 
               
               
                   
                   
                 NO: 6 or 7) + 
               
               
                   
                   
                 Amylase (SEQ ID 
               
               
                   
                   
                 NO: 5) 
               
               
                 XX H 1-4 
                 Differs 
                 Protease (SEQ ID 
                 X 
                 X 
                 X 
               
               
                   
                   
                 NO: 6 or 7) + 
               
               
                   
                   
                 Amylase (SEQ ID 
               
               
                   
                   
                 NO: 5) 
               
               
                   
               
            
           
         
       
     
     E.g. Labels for control was called None, and labeled NO A1-4 etc. 
     Egg=EG, Broccoli=BR, Fish=FI. When the foodstuff is egg the protease of SEQ ID NO: 6 is used. When the foodstuff is not egg the protease of SEQ ID NO: 7 is used. 
     Odor Panel evaluation of Results (Assay II). The odor panel this time was asked to rank the odor from 1 to 5, where 5 was the most pungent odor, and 1 was not significantly different from the blank odor. The odor panel consisted of 5 trained panelists. Data can be found in table 9. Data from Assay I can be found in tables 6-8. 
     The foodstuffs were prepared and processed as described above in paragraphs “Preparation of foodstuffs for odor generation” and “Processing of foodstuff after initial preparation” and with the experimental setup describe above. One experiment is performed with a commercial detergent composition instead of buffer, where 1 tablet of SUN prof dishwash tablets P K100 HG756 is dissolved in 5 L water instead of buffer. The composition of SUN prof dishwash tablets P K100 HG756 is: 5-15% oxygen based bleaching agents and less than 5% non-ionic surfactant, phosphonates and polycarboxylates. The tablets were used to determine a fragrance masking profile instead of an odor removal. No enzymes were added to the tablet prepared foodstuffs. 
     
       
         
           
               
             
               
                 TABLE 6 
               
               
                   
               
               
                 Assay I: GC Headspace retention measurements for Broccoli with different enzymes. Channel 1, a cutoff 
               
               
                 value of 5,000 was chosen to pick the most relevant peaks, similar trends were seen in channel 2 (data not shown). 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                   
                 13.01- 
                 14.39- 
                 15.66- 
                 16.00- 
                 17.19- 
                 18.36- 
                 19.50- 
                 22.65- 
               
               
                 GC HeadSpace Retention Time-Channel 1 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
               
               
                   
               
               
                 Broccoli in Buffer background 
                 3915 
                 53666 
                 0 
                 1177 
                 10023 
                 23594 
                 0 
                 1800 
               
               
                 Broccoli + protease and amylase 
                 6434 
                 0 
                 0 
                 3497 
                 5988 
                 17821 
                 0 
                 1791 
               
               
                 Broccoli + protease, amylase, mannanase 
                 7727 
                 17077 
                 0 
                 2520 
                 6520 
                 17532 
                 0 
                 4284 
               
               
                 Broccoli + protease, amylase, cellulose 
                 5860 
                 46750 
                 0 
                 1409 
                 13000 
                 23542 
                 0 
                 1810 
               
               
                 Broccoli + protease, amylase, oxidase 
                 6938 
                 4823 
                 0 
                 11650 
                 6411 
                 16323 
                 0 
                 8792 
               
               
                 Broccoli + protease, amylase, cellulase, 
                 7212 
                 0 
                 0 
                 3366 
                 5200 
                 15193 
                 0 
                 4937 
               
               
                 mannanase 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Broccoli + protease, amylase, cellulase, oxidase  
                 5708 
                 4276 
                 0 
                 10305 
                 5903 
                 15104 
                 5123 
                 8074 
               
               
                 Broccoli + protease, amylase, mannanase, 
                 7342 
                 5111 
                 0 
                 10854 
                 5613 
                 15913 
                 0 
                 7457 
               
               
                 oxidase 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Broccoli + protease, amylase, mannanase, 
                 5545 
                 4382 
                 0 
                 8827 
                 4076 
                 15143 
                 0 
                 6055 
               
               
                 cellulase, oxidase 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Broccoli in Tablet 
                 11730 
                 0 
                 0 
                 3458 
                 3311 
                 17117 
                 0 
                 4146 
               
               
                   
               
               
                   
                 37.30- 
                 45.74- 
                 47.21- 
                 48.91- 
                 51.53- 
                 58.07- 
                 62.11- 
                 77.04- 
               
               
                 GC HeadSpace Retention Time-Channel 1 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
               
               
                   
               
               
                 Broccoli in Buffer background 
                 5157 
                 42 
                 18913 
                 1200 
                 7282 
                 2511 
                 6364 
                 3580 
               
               
                 Broccoli + protease and amylase 
                 4935 
                 5482 
                 0 
                 2230 
                 5047 
                 2033 
                 6491 
                 2596 
               
               
                 Broccoli + protease, amylase, mannanase 
                 40159 
                 20160 
                 0 
                 5051 
                 4255 
                 3081 
                 6879 
                 1891 
               
               
                 Broccoli + protease, amylase, cellulase 
                 4961 
                 6394 
                 0 
                 2576 
                 5160 
                 1735 
                 7215 
                 1751 
               
               
                 Broccoli + protease, amylase, oxidase 
                 50814 
                 6079 
                 0 
                 2339 
                 2270 
                 2525 
                 7735 
                 2425 
               
               
                 Broccoli + protease, amylase, cellulase, 
                 51099 
                 18209 
                 0 
                 4644 
                 3494 
                 3867 
                 7190 
                 2003 
               
               
                 mannanase 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Broccoli + protease, amylase, cellulase, oxidase 
                 46165 
                 8606 
                 0 
                 2841 
                 2296 
                 2458 
                 7423 
                 2141 
               
               
                 Broccoli + protease, amylase, mannanase, 
                 40817 
                 16123 
                 0 
                 4439 
                 3013 
                 2805 
                 8335 
                 2182 
               
               
                 oxidase 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Broccoli + protease, amylase, mannanase, 
                 32176 
                 7514 
                 0 
                 2509 
                 1741 
                 1890 
                 7740 
                 1639 
               
               
                 cellulase, oxidase 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Broccoli in Tablet 
                 85573 
                 1651 
                 0 
                 1515 
                 1818 
                 7252 
                 7929 
                 9835 
               
               
                   
               
               
                   
                 96.20- 
                 99.24- 
                 104.97- 
                 107.91- 
                 112.32- 
                 118.04- 
                 128.85- 
                 133.99- 
               
               
                 GC HeadSpace Retention Time-Channel 1 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
               
               
                   
               
               
                 Broccoli in Buffer background 
                 131 
                 760 
                 1423 
                 1266 
                 25 
                 644 
                 1261 
                 285 
               
               
                 Broccoli + protease and amylase 
                 169 
                 609 
                 770 
                 1127 
                 0 
                 640 
                 4879 
                 273 
               
               
                 Broccoli + protease, amylase, mannanase 
                 517 
                 675 
                 546 
                 979 
                 0 
                 533 
                 16843 
                 120 
               
               
                 Broccoli + protease, amylase, cellulase 
                 226 
                 640 
                 641 
                 851 
                 0 
                 492 
                 6784 
                 396 
               
               
                 Broccoli + protease, amylase, oxidase 
                 168 
                 774 
                 532 
                 964 
                 0 
                 429 
                 6600 
                 248 
               
               
                 Broccoli + protease, amylase, cellulase, 
                 453 
                 1002 
                 649 
                 854 
                 0 
                 536 
                 12515 
                 190 
               
               
                 mannanase 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Broccoli + protease, amylase, cellulase, oxidase 
                 143 
                 961 
                 487 
                 828 
                 0 
                 446 
                 7033 
                 113 
               
               
                 Broccoli + protease, amylase, mannanase, 
                 250 
                 942 
                 1654 
                 1027 
                 193 
                 693 
                 16334 
                 265 
               
               
                 oxidase 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Broccoli + protease, amylase, mannanase, 
                 29 
                 1024 
                 405 
                 904 
                 0 
                 493 
                 11464 
                 181 
               
               
                 cellulase, oxidase 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Broccoli in Tablet 
                 38638 
                 71296 
                 179156 
                 18459 
                 24561 
                 23836 
                 21949 
                 5004 
               
               
                   
               
            
           
           
               
               
               
               
               
               
            
               
                   
                 141.96- 
                 147.41- 
                 155.13- 
                 159.93- 
                 191.32- 
               
               
                 GC HeadSpace Retention Time-Channel 1 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
               
               
                   
               
               
                 Broccoli in Buffer background 
                 238 
                 408 
                 1762 
                 1533 
                 7718 
               
               
                 Broccoli + protease and amylase 
                 155 
                 47 
                 1704 
                 1582 
                 7358 
               
               
                 Broccoli + protease, amylase, mannanase 
                 139 
                 0 
                 1801 
                 1696 
                 7348 
               
               
                 Broccoli + protease, amylase, cellulase 
                 177 
                 0 
                 1948 
                 1628 
                 7205 
               
               
                 Broccoli + protease, amylase, oxidase 
                 110 
                 0 
                 1521 
                 1795 
                 6923 
               
               
                 Broccoli + protease, amylase, cellulase, 
                 41 
                 0 
                 1641 
                 1838 
                 6770 
               
               
                 mannanase 
                   
                   
                   
                   
                   
               
               
                 Broccoli + protease, amylase, cellulase, oxidase 
                 72 
                 0 
                 1509 
                 1983 
                 6351 
               
               
                 Broccoli + protease, amylase, mannanase, 
                 159 
                 631 
                 1667 
                 2051 
                 6372 
               
               
                 oxidase 
                   
                   
                   
                   
                   
               
               
                 Broccoli + protease, amylase, mannanase, 
                 103 
                 0 
                 1391 
                 1921 
                 6351 
               
               
                 cellulase, oxidase 
                   
                   
                   
                   
                   
               
               
                 Broccoli in Tablet 
                 7863 
                 36287 
                 15596 
                 6251 
                 6682 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 7 
               
               
                   
               
               
                 Assay I: GC Headspace retention measurements for Egg with different enzymes. Channel 1, a cutoff value  
               
               
                 of 5,000 was chosen to pick the most relevant peaks, similar trends were seen in channel 2 (data not shown). 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                 GC HeadSpace Retention 
                 14.48- 
                 15.92- 
                 18.46- 
                 20.01- 
                 21.54- 
                 30.34- 
                 45.00- 
                 46.08- 
               
               
                 Time-Channel 1 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
               
               
                   
               
               
                 Egg in Buffer background 
                 38632 
                 0 
                 11091 
                 0 
                 0 
                 907 
                 5961 
                 1735 
               
               
                 Egg + protease and amylase 
                 0 
                 6322 
                 9377 
                 0 
                 0 
                 169 
                 0 
                 12917 
               
               
                 Egg + protease, amylase, mannanase 
                 3571 
                 527675 
                 7125 
                 92912 
                 24445 
                 131 
                 0 
                 29410 
               
               
                 Egg + protease, amylase, cellulase 
                 0 
                 10833 
                 9090 
                 0 
                 0 
                 188 
                 0 
                 8895 
               
               
                 Egg + protease, amylase, oxidase 
                 2845 
                 10685 
                 10328 
                 0 
                 0 
                 818 
                 0 
                 21041 
               
               
                 Egg + protease, amylase, cellulase,  
                 0 
                 5619 
                 9191 
                 0 
                 0 
                 325 
                 0 
                 37567 
               
               
                 mannanase 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Egg + protease, amylase, cellulase,  
                 2576 
                 9205 
                 9836 
                 0 
                 0 
                 895 
                 0 
                 9481 
               
               
                 oxidase 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Egg + protease, amylase, mannanase,  
                 3245 
                 11251 
                 9699 
                 0 
                 0 
                 1981 
                 0 
                 16306 
               
               
                 oxidase 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Egg + protease, amylase, mannanase,  
                 2893 
                 10498 
                 10772 
                 0 
                 0 
                 1838 
                 0 
                 13809 
               
               
                 cellulase, oxidase 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Egg in Tablet 
                 0 
                 3246 
                 13446 
                 0 
                 0 
                 6092 
                 288 
                 1459 
               
               
                   
               
               
                 GC HeadSpace Retention  
                 47.14- 
                 49.15- 
                 58.37- 
                 60.44- 
                 62.36- 
                 65.21- 
                 77.36- 
                 84.25- 
               
               
                 Time-Channel 1 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
               
               
                   
               
               
                 Egg in Buffer background 
                 0 
                 1396 
                 2437 
                 0 
                 7744 
                 0 
                 649 
                 80 
               
               
                 Egg + protease and amylase 
                 0 
                 4334 
                 1634 
                 0 
                 8282 
                 0 
                 668 
                 71 
               
               
                 Egg + protease, amylase, mannanase 
                 42767 
                 7722 
                 4551 
                 2431 
                 7333 
                 34602 
                 797 
                 105 
               
               
                 Egg + protease, amylase, cellulase 
                 0 
                 2124 
                 1770 
                 0 
                 7086 
                 122 
                 611 
                 70 
               
               
                 Egg + protease, amylase, oxidase 
                 0 
                 5478 
                 3215 
                 48 
                 7646 
                 0 
                 1638 
                 118 
               
               
                 Egg + protease, amylase, cellulase,  
                 0 
                 9401 
                 5414 
                 186 
                 8230 
                 0 
                 721 
                 89 
               
               
                 mannanase 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Egg + protease, amylase, cellulase, 
                 0 
                 3220 
                 3616 
                 0 
                 8280 
                 0 
                 1737 
                 105 
               
               
                 oxidase 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Egg +protease, amylase, mannanase,  
                 0 
                 4801 
                 4761 
                 19 
                 8354 
                 0 
                 2002 
                 152 
               
               
                 oxidase 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Egg + protease, amylase, mannanase,  
                 0 
                 3966 
                 4300 
                 22 
                 8435 
                 0 
                 1712 
                 124 
               
               
                 cellulase, oxidase 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Egg in Tablet 
                 0 
                 1233 
                 17270 
                 0 
                 8451 
                 0 
                 14501 
                 8393 
               
               
                   
               
               
                 GC HeadSpace Retention  
                 87.21- 
                 92.49- 
                 97.13- 
                 99.40- 
                 104.08- 
                 109.05- 
                 112.62- 
                 128.48- 
               
               
                 Time-Channel 1 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
               
               
                   
               
               
                 Egg in Buffer background 
                 1079 
                 6833 
                 180 
                 754 
                 513 
                 3559 
                 0 
                 1360 
               
               
                 Egg + protease and amylase 
                 851 
                 6078 
                 186 
                 651 
                 293 
                 3644 
                 0 
                 6165 
               
               
                 Egg + protease, amylase, mannanase 
                 820 
                 5667 
                 137 
                 646 
                 381 
                 3851 
                 0 
                 22389 
               
               
                 Egg + protease, amylase, cellulase 
                 728 
                 5099 
                 135 
                 632 
                 170 
                 3086 
                 0 
                 4474 
               
               
                 Egg + protease, amylase, oxidase 
                 682 
                 5711 
                 146 
                 800 
                 198 
                 3565 
                 0 
                 7306 
               
               
                 Egg + protease, amylase, cellulase,  
                 746 
                 5505 
                 120 
                 408 
                 242 
                 3271 
                 0 
                 14595 
               
               
                 mannanase 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Egg + protease, amylase, cellulase,  
                 595 
                 4825 
                 126 
                 707 
                 257 
                 3464 
                 0 
                 6529 
               
               
                 oxidase 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Egg + protease, amylase, mannanase,  
                 613 
                 5037 
                 176 
                 777 
                 554 
                 3541 
                 0 
                 15227 
               
               
                 oxidase 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Egg + protease, amylase, mannanase,  
                 676 
                 5260 
                 143 
                 592 
                 265 
                 3478 
                 0 
                 13035 
               
               
                 cellulase, oxidase 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Egg in Tablet 
                 5464 
                 6025 
                 16888 
                 37795 
                 63292 
                 8227 
                 7257 
                 4788 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 8 
               
               
                   
               
               
                 Assay I: GC Headspace retention measurements for Fish with different enzymes. Channel 1, a cutoff value  
               
               
                 of 5,000 was chosen to pick the most relevant peaks, similar trends were seen in channel 2 (data not shown). 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                 GC HeadSpace Retention Time-  
                 13.01- 
                 13.56- 
                 13.70- 
                 14.39- 
                 15.66- 
                 16.00- 
                 17.19- 
                 18.36- 
               
               
                 Channel 1 Run 1 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
               
               
                   
               
               
                 Fish in Buffer background 
                 9296 
                 0 
                 0 
                 635592 
                 0 
                 0 
                 0 
                 12526 
               
               
                 Fish + protease and amylase 
                 4265 
                 0 
                 0 
                 4506 
                 0 
                 28304 
                 0 
                 11699 
               
               
                 Fish + protease, amylase, mannanase 
                 5281 
                 0 
                 0 
                 2491 
                 0 
                 26373 
                 0 
                 11487 
               
               
                 Fish + protease, amylase, cellulase 
                 4534 
                 0 
                 0 
                 2788 
                 0 
                 27315 
                 0 
                 11161 
               
               
                 Fish + protease, amylase, oxidase 
                 4186 
                 0 
                 0 
                 5074 
                 0 
                 41757 
                 0 
                 12514 
               
               
                 Fish + protease, amylase, cellulase,  
                 4714 
                 0 
                 0 
                 2738 
                 0 
                 25644 
                 0 
                 11110 
               
               
                 mannanase 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Fish + protease, amylase, cellulase,  
                 4023 
                 0 
                 0 
                 3863 
                 0 
                 36836 
                 0 
                 11754 
               
               
                 oxidase 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Fish + protease, amylase, mannanase,  
                 5267 
                 0 
                 4799 
                 42042 
                 0 
                 0 
                 11856 
                 0 
               
               
                 oxidase 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Fish + protease, amylase, mannanase,  
                 5116 
                 0 
                 5332 
                 41949 
                 0 
                 0 
                 11238 
                 0 
               
               
                 cellulase, oxidase 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Fish in Tablet 
                 3913 
                 60723 
                 91147 
                 9987 
                 0 
                 0 
                 7037 
                 3240 
               
               
                   
               
               
                 GC HeadSpace Retention Time-  
                 19.50- 
                 20.45- 
                 21.32- 
                 26.48- 
                 44.72- 
                 45.74- 
                 47.21- 
                 48.91- 
               
               
                 Channel 1 Run 1 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
               
               
                   
               
               
                 Fish in Buffer background 
                 44119 
                 0 
                 0 
                 66221 
                 50565 
                 0 
                 0 
                 2961 
               
               
                 Fish + protease and amylase 
                 0 
                 16425 
                 0 
                 197 
                 0 
                 2748 
                 0 
                 2538 
               
               
                 Fish + protease, amylase, mannanase 
                 0 
                 17547 
                 0 
                 131 
                 0 
                 57364 
                 0 
                 18345 
               
               
                 Fish + protease, amylase, cellulase 
                 0 
                 16274 
                 0 
                 97 
                 0 
                 10419 
                 0 
                 4346 
               
               
                 Fish + protease, amylase, oxidase 
                 0 
                 17190 
                 0 
                 192 
                 0 
                 6194 
                 0 
                 3387 
               
               
                 Fish + protease, amylase, cellulase,  
                 0 
                 16604 
                 0 
                 71 
                 0 
                 45311 
                 0 
                 11305 
               
               
                 mannanase 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Fish + protease, amylase, cellulase,  
                 0 
                 16498 
                 0 
                 148 
                 0 
                 7960 
                 0 
                 3198 
               
               
                 oxidase 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Fish + protease, amylase, mannanase,  
                 0 
                 0 
                 17456 
                 0 
                 0 
                 0 
                 44917 
                 0 
               
               
                 oxidase 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Fish + protease, amylase, mannanase,  
                 0 
                 0 
                 16729 
                 0 
                 0 
                 0 
                 83758 
                 0 
               
               
                 cellulase, oxidase 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Fish in Tablet 
                 0 
                 0 
                 7851 
                 796 
                 574 
                 349 
                 1356 
                 0 
               
               
                   
               
               
                 GC HeadSpace Retention Time-  
                 51.53- 
                 58.07- 
                 59.78- 
                 62.11- 
                 87.04- 
                 90.62- 
                 92.23- 
                 96.20- 
               
               
                 Channel 1 Run 1 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
               
               
                   
               
               
                 Fish in Buffer background 
                 609 
                 2082 
                 0 
                 7223 
                 579 
                 169 
                 9509 
                 5374 
               
               
                 Fish + protease and amylase 
                 266 
                 4042 
                 0 
                 7557 
                 617 
                 181 
                 8238 
                 5861 
               
               
                 Fish + protease, amylase, mannanase 
                 0 
                 5715 
                 0 
                 9348 
                 737 
                 0 
                 9509 
                 7489 
               
               
                 Fish + protease, amylase, cellulase 
                 638 
                 4243 
                 16 
                 7701 
                 612 
                 139 
                 8296 
                 5898 
               
               
                 Fish + protease, amylase, oxidase 
                 467 
                 4996 
                 0 
                 7628 
                 621 
                 172 
                 9503 
                 5466 
               
               
                 Fish + protease, amylase, cellulase,  
                 531 
                 4708 
                 0 
                 8292 
                 592 
                 34 
                 9040 
                 5745 
               
               
                 mannanase 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Fish + protease, amylase, cellulase,  
                 346 
                 5152 
                 0 
                 7991 
                 656 
                 134 
                 8317 
                 5757 
               
               
                 oxidase 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Fish + protease, amylase, mannanase,  
                 10484 
                 5123 
                 9086 
                 0 
                 1579 
                 2260 
                 853 
                 12577 
               
               
                 oxidase 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Fish + protease, amylase, mannanase,  
                 20010 
                 6025 
                 9714 
                 0 
                 1628 
                 1245 
                 655 
                 11711 
               
               
                 cellulase, oxidase 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Fish in Tablet 
                 1497 
                 6253 
                 8079 
                 0 
                 10189 
                 7449 
                 1011 
                 13312 
               
               
                   
               
               
                 GC HeadSpace Retention Time-  
                 99.24- 
                 104.97- 
                 107.91- 
                 109.57- 
                 110.92- 
                 112.32- 
                 114.46- 
                 115.97- 
               
               
                 Channel 1 Run 1 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
               
               
                   
               
               
                 Fish in Buffer background 
                 6238 
                 579 
                 1178 
                 2081 
                 1474 
                 0 
                 1690 
                 0 
               
               
                 Fish + protease and amylase 
                 6528 
                 435 
                 1321 
                 5110 
                 993 
                 0 
                 1569 
                 0 
               
               
                 Fish + protease, amylase, mannanase 
                 7290 
                 471 
                 1207 
                 6155 
                 1070 
                 0 
                 1785 
                 0 
               
               
                 Fish + protease, amylase, cellulase 
                 6568 
                 381 
                 1080 
                 5815 
                 1051 
                 0 
                 1540 
                 0 
               
               
                 Fish + protease, amylase, oxidase 
                 6541 
                 369 
                 1360 
                 6281 
                 981 
                 0 
                 1766 
                 0 
               
               
                 Fish + protease, amylase, cellulase,  
                 6497 
                 371 
                 1220 
                 6447 
                 1087 
                 0 
                 1868 
                 0 
               
               
                 mannanase 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Fish + protease, amylase, cellulase,  
                 6469 
                 432 
                 1293 
                 6226 
                 1007 
                 0 
                 1604 
                 0 
               
               
                 oxidase 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Fish + protease, amylase, mannanase,  
                 1772 
                 5866 
                 6918 
                 861 
                 0 
                 9709 
                 0 
                 1998 
               
               
                 oxidase 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Fish + protease, amylase, mannanase,  
                 1780 
                 5536 
                 6796 
                 1429 
                 22 
                 8771 
                 0 
                 2034 
               
               
                 cellulase, oxidase 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Fish in Tablet 
                 1758 
                 34972 
                 73209 
                 139866 
                 12731 
                 19184 
                 10799 
                 9779 
               
               
                   
               
               
                 GC HeadSpace Retention Time-  
                 118.04- 
                 124.88- 
                 128.85- 
                 141.96- 
                 147.41- 
                 149.51- 
                 155.13- 
                 162.18- 
               
               
                 Channel 1 Run 1 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
               
               
                   
               
               
                 Fish in Buffer background 
                 226 
                 63 
                 1458 
                 697 
                 15 
                 2811 
                 8116 
                 385 
               
               
                 Fish + protease and amylase 
                 429 
                 27 
                 2923 
                 989 
                 0 
                 2835 
                 8326 
                 377 
               
               
                 Fish + protease, amylase, mannanase 
                 717 
                 27 
                 39550 
                 905 
                 0 
                 2764 
                 8562 
                 343 
               
               
                 Fish + protease, amylase, cellulase 
                 752 
                 0 
                 8142 
                 947 
                 0 
                 2635 
                 8431 
                 354 
               
               
                 Fish + protease, amylase, oxidase 
                 927 
                 0 
                 5442 
                 891 
                 0 
                 2899 
                 8694 
                 381 
               
               
                 Fish + protease, amylase, cellulase,  
                 1144 
                 13 
                 42090 
                 909 
                 0 
                 2885 
                 8220 
                 338 
               
               
                 mannanase 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Fish + protease, amylase, cellulase,  
                 1139 
                 0 
                 7698 
                 911 
                 0 
                 2749 
                 8249 
                 337 
               
               
                 oxidase 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Fish + protease, amylase, mannanase,  
                 2384 
                 32415 
                 808 
                 1459 
                 10897 
                 0 
                 3133 
                 8289 
               
               
                 oxidase 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Fish + protease, amylase, mannanase,  
                 2446 
                 38881 
                 522 
                 1316 
                 6232 
                 89 
                 2898 
                 7736 
               
               
                 cellulase, oxidase 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Fish in Tablet 
                 12569 
                 11195 
                 1591 
                 5279 
                 8162 
                 8600 
                 3309 
                 11077 
               
               
                   
               
            
           
           
               
               
               
            
               
                 GC HeadSpace Retention Time-  
                 191.32- 
                 204.95- 
               
               
                 Channel 1 Run 1 
                 1-A 
                 1-A 
               
               
                   
               
               
                 Fish in Buffer background 
                 6544 
                 62 
               
               
                 Fish + protease and amylase 
                 6502 
                 83 
               
               
                 Fish + protease, amylase, mannanase 
                 6819 
                 67 
               
               
                 Fish + protease, amylase, cellulose 
                 7008 
                 84 
               
               
                 Fish + protease, amylase, oxidase 
                 6784 
                 63 
               
               
                 Fish + protease, amylase, cellulase,  
                 6876 
                 81 
               
               
                 mannanase 
                   
                   
               
               
                 Fish + protease, amylase, cellulase,  
                 7756 
                 84 
               
               
                 oxidase 
                   
                   
               
               
                 Fish + protease, amylase, mannanase,  
                 561 
                 6248 
               
               
                 oxidase 
                   
                   
               
               
                 Fish + protease, amylase, mannanase,  
                 523 
                 7001 
               
               
                 cellulase, oxidase 
                   
                   
               
               
                 Fish in Tablet 
                 603 
                 6663 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 9 
               
             
            
               
                   
               
               
                 Smell Panel Evaluation 
               
            
           
           
               
               
               
               
               
               
            
               
                 PANELLIST 
                 1 
                 2 
                 3 
                 4 
                 5 
               
               
                   
               
            
           
           
               
               
            
               
                   
                 BROCCOLI 
               
            
           
           
               
               
               
               
               
               
            
               
                 Food in Buffer control 
                 5.0 
                 4.0 
                 3.0 
                 3.0 
                 5.0 
               
               
                 Food + protease and amylase 
                 3.0 
                 4.0 
                 4.0 
                 3.0 
                 3.0 
               
               
                 Food + protease, amylase, mannanase 
                 2.0 
                 4.0 
                 5.0 
                 1.0 
                 2.0 
               
               
                 Food + protease, amylase, cellulase 
                 3.0 
                 4.0 
                 5.0 
                 2.0 
                 1.0 
               
               
                 Food + protease, amylase, oxidase 
                 2.0 
                 3.0 
                 2.0 
                 1.0 
                 1.0 
               
               
                 Food + protease, amylase, cellulase, 
                 2.0 
                 3.0 
                 1.0 
                 2.0 
                 5.0 
               
               
                 mannanase 
               
               
                 Food + protease, amylase, cellulase, 
                 1.5 
                 2.0 
                 1.0 
                 2.0 
                 1.0 
               
               
                 oxidase 
               
               
                 Food + protease, amylase, mannanase, 
                 2.0 
                 1.0 
                 2.0 
                 1.0 
                 2.0 
               
               
                 oxidase 
               
               
                 Food + protease, amylase, mannanase, 
                 1.5 
                 1.0 
                 1.0 
                 1.0 
                 1.0 
               
               
                 cellulase, oxidase 
               
            
           
           
               
               
            
               
                   
                 FISH 
               
            
           
           
               
               
               
               
               
               
            
               
                 Food in Buffer control 
                 5.0 
                 4.0 
                 4.0 
                 5.0 
                 5.0 
               
               
                 Food + protease and amylase 
                 4.0 
                 3.0 
                 2.0 
                 5.0 
                 4.0 
               
               
                 Food + protease, amylase, mannanase 
                 3.5 
                 3.0 
                 1.0 
                 5.0 
                 4.0 
               
               
                 Food + protease, amylase, cellulase 
                 3.0 
                 4.0 
                 1.0 
                 4.0 
                 2.0 
               
               
                 Food + protease, amylase, oxidase 
                 2.5 
                 3.0 
                 1.0 
                 3.0 
                 2.0 
               
               
                 Food + protease, amylase, cellulase, 
                 3.0 
                 4.0 
                 1.0 
                 1.0 
                 1.0 
               
               
                 mannanase 
               
               
                 Food + protease, amylase, cellulase, 
                 2.0 
                 1.0 
                 1.0 
                 1.0 
                 1.0 
               
               
                 oxidase 
               
               
                 Food + protease, amylase, mannanase, 
                 1.5 
                 1.0 
                 1.0 
                 1.0 
                 1.0 
               
               
                 oxidase 
               
               
                 Food + protease, amylase, mannanase, 
                 3.0 
                 2.0 
                 1.0 
                 1.0 
                 1.0 
               
               
                 cellulase, oxidase 
               
            
           
           
               
               
            
               
                   
                 EGG 
               
            
           
           
               
               
               
               
               
               
            
               
                 Food in Buffer control 
                 3.0 
                 4.0 
                 3.0 
                 5.0 
                 4.0 
               
               
                 Food + protease and amylase 
                 5.0 
                 3.0 
                 5.0 
                 5.0 
                 4.0 
               
               
                 Food + protease, amylase, mannanase 
                 2.0 
                 3.0 
                 2.0 
                 5.0 
                 3.0 
               
               
                 Food + protease, amylase, cellulase 
                 2.5 
                 4.0 
                 3.0 
                 5.0 
                 3.0 
               
               
                 Food + protease, amylase, oxidase 
                 1.0 
                 4.0 
                 2.0 
                 5.0 
                 2.0 
               
               
                 Food + protease, amylase, cellulase, 
                 3.5 
                 3.0 
                 1.0 
                 4.0 
                 4.0 
               
               
                 mannanase 
               
               
                 Food + protease, amylase, cellulase, 
                 2.0 
                 1.0 
                 1.0 
                 1.0 
                 4.0 
               
               
                 oxidase 
               
               
                 Food + protease, amylase, mannanase, 
                 2.0 
                 2.0 
                 2.0 
                 1.0 
                 4.0 
               
               
                 oxidase 
               
               
                 Food + protease, amylase, mannanase, 
                 1.0 
                 2.0 
                 1.0 
                 1.0 
                 4.0 
               
               
                 cellulase, oxidase 
               
               
                   
               
            
           
         
       
     
     Example 4. Full Scale Washing 
     The five foodstuffs (Broccoli, Egg, Fish, Onion, and Garlic) were prepared as described under examples under Preparation of foodstuffs for odor generation. 
     Processing of Foodstuff after Initial Preparation. 
     Assay Buffer: Buffer pH 8.00. 50 mM HEPES, 50 mM CHES, 1 mM CaCl 2 ), 0.01% triton X-100. 
     Water hardness at 21° dH was added to the buffer. The Ca:NaHCO 3 :Mg ratio was 4:2:1, prepared using 4000° dH CaCl 2 ), 2000° dH MgCl2 stock solutions and freshly prepared 0.535 M NaHCO 3 . 
     After this initial preparation, 4×200 mL bottles of buffer were aliquoted. 12.5 g of broccoli, 12.5 g of Fish, 12.5 g of egg, 2 g of onion, and 1 g of garlic was added to each of the four bottles. These were stirred at 500 rpm on a magnetic stirrer for 20 min to allow for homogeneity. These bottles of mixed foods were labelled A, B, C, D. 
     To bottles A and C, the enzymes according to the experimental set up is shown tables 1-2 above was added to the same end concentration. The enzyme were stirred in the food mix for 5 min. Bottles A, B, C, and D were added to the filter of four Miele GLS2 dishwashing machines. Machines with A and B were then run through a wash program (only with water) immediately at 40° C., using the standard program. R 40° C., 8′/20′/55′ KL 55° C. Machines with C And D were closed and not washed. All four machines were then left for 72 h. After this period all four were run through the wash program with water again, and a panel evaluated the odour of each immediately after the drying phase completion. 
     The odor panel consists of 5 trained panelists. Each machine was opened, and all 5 panelists performed olfactory evaluation simultaneously. This was repeated for all four machines. The panelists were then asked to compare A vs B and then C vs D, stating which machine had “won” as a ranking based on odour reduction. 
     
       
         
           
               
             
               
                 TABLE 10 
               
             
            
               
                   
               
               
                 Full Scale Wash Evaluation of Odor in Dishwasher Machines, Those 
               
               
                 washed immediately and then left for 72 h, and then washed again. 
               
            
           
           
               
               
               
            
               
                 Panellists 
                 A 
                 B 
               
               
                   
               
               
                 1 
                 Win 
                 Lose 
               
               
                 2 
                 Win 
                 Lose 
               
               
                 3 
                 Win 
                 Lose 
               
               
                 4 
                 Win 
                 Lose 
               
               
                 5 
                 Win 
                 Lose 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 11 
               
             
            
               
                   
               
               
                 Full Scale Wash Evaluation of Odor in Dishwasher 
               
               
                 Machines, Those left for 72 h, and then washed. 
               
            
           
           
               
               
               
            
               
                 Panellists 
                 C 
                 D 
               
               
                   
               
               
                 1 
                 Win 
                 lose 
               
               
                 2 
                 Lose 
                 Win 
               
               
                 3 
                 Win 
                 Lose 
               
               
                 4 
                 Lose 
                 Win 
               
               
                 5 
                 Win 
                 Lose 
               
               
                   
               
            
           
         
       
     
     Example 5. MDW Odor Removal 
     Determination of the Necessary Enzymes for Odor Removal. 
     Fresh onions were purchased from a local supermarket (Netto, Bagsværd) and were used untreated and raw after peeling. 
     Processing of Substrates after Initial Preparation. 
     Assay Buffer: Buffer pH 8.00. 50 mM HEPES, 50 mM CHES, 1 mM CaCl 2 ), 0.01% triton X-100. 
     Water hardness at 21° dH was added to the buffer. The Ca:NaHCO 3 :Mg ratio was 4:2:1, prepared using 4000° dH CaCl 2 ), 2000° dH MgCl2 stock solutions and freshly prepared 0.535 M NaHCO 3 . 
     Onion Preparation:
         1 g of blended substrate was added to 100 mL of assay buffer and mixed to a fine liquid suspension (ca. 3 min of blending).   2 replicates of 2 mL were pipetted to the gas E-nose vials.   The substrate was prepared first and after pipetting the lids screwed tightly in place.   The vials were mixed using a whirl mixer for 30 s.   Controls are listed below.   All vials were incubated at 37° C. over 24 h.   E-Nose analysis (Assay I) was carried out and SAS JMP used to perform a principle component analysis of the data to determine which enzymes play a critical role.       

     
       
         
           
               
             
               
                 TABLE 12 
               
             
            
               
                   
               
               
                 Design, an X indicates the addition n of the enzyme 
               
               
                 to the vial as per table 2 concentration. 
               
            
           
           
               
               
               
               
               
               
               
            
               
                   
                   
                 SEQ ID 
                 SEQ ID 
                 SEQ ID 
                 SEQ ID 
                 SEQ ID 
               
               
                   
                 Ref# 
                 NO: 7 
                 NO: 5 
                 NO: 1 
                 NO: 2 
                 NO: 4 
               
               
                   
                   
               
               
                   
                  1/31 
                 X 
                   
                   
                   
                   
               
               
                   
                  2/32 
                   
                 X 
               
               
                   
                  3/33 
                   
                   
                 X 
               
               
                   
                  4/34 
                   
                   
                   
                 x 
               
               
                   
                  5/35 
                   
                   
                   
                   
                 X 
               
               
                   
                  6/36 
                 X 
                 X 
               
               
                   
                  7/37 
                 X 
                   
                 X 
               
               
                   
                  8/38 
                 X 
                   
                   
                 x 
               
               
                   
                  9/39 
                 X 
                   
                   
                   
                 X 
               
               
                   
                 10/40 
                   
                 x 
                 X 
               
               
                   
                 11/41 
                   
                 x 
                   
                 x 
               
               
                   
                 12/42 
                   
                 x 
                   
                   
                 X 
               
               
                   
                 13/43 
                   
                   
                 X 
                 x 
               
               
                   
                 14/44 
                   
                   
                 X 
                   
                 X 
               
               
                   
                 15/45 
                   
                   
                   
                 x 
                 X 
               
               
                   
                 16/46 
                 X 
                 x 
                 X 
               
               
                   
                 17/47 
                 X 
                   
                 X 
                 x 
               
               
                   
                 18/48 
                 X 
                   
                   
                 x 
                 X 
               
               
                   
                 19/49 
                   
                 x 
                 X 
                 x 
               
               
                   
                 20/50 
                   
                 x 
                 X 
                   
                 X 
               
               
                   
                 21/51 
                   
                   
                 X 
                 x 
                 X 
               
               
                   
                 22/52 
                 X 
                 x 
                   
                 x 
               
               
                   
                 23/53 
                 X 
                 x 
                   
                   
                 X 
               
               
                   
                 24/54 
                   
                 x 
                   
                 x 
                 X 
               
               
                   
                 25/55 
                 X 
                 x 
                 X 
                 x 
               
               
                   
                 26/56 
                   
                 x 
                 X 
                 x 
                 X 
               
               
                   
                 27/57 
                 X 
                   
                 X 
                 x 
                 X 
               
               
                   
                 28/58 
                 X 
                 x 
                   
                 x 
                 X 
               
               
                   
                 29/59 
                 X 
                 x 
                 X 
                   
                 X 
               
               
                   
                 30/60 
                 X 
                 x 
                 X 
                 x 
                 X 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 13 
               
               
                   
               
               
                 E-nose retention measurements (Assay I) for food items with and without enzymes. Channel 1, a cutoff  
               
               
                 value of 5,000 was chosen to pick the most relevant peaks, similar trends were seen in channel 2 
               
               
                 (data not shown). Where P = protease (SEQ ID NO: 7), A = amylase (SEQ ID NO: 5), C = cellulase 
               
               
                 (SEQ ID NO: 1), M = mannanase (SEQ ID NO: 2), O = cellobiose oxidase (SEQ ID NO: 4). 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                   
                 13.07-1- 
                 13.80-1- 
                 14.52-1- 
                 15.86-1- 
                 18.49-1- 
                 19.78-1- 
                 20.75-1- 
                 21.20-1- 
               
               
                   
                 A 
                 A 
                 A 
                 A 
                 A 
                 A 
                 A 
                 A 
               
               
                   
               
               
                 Onion + P 
                 1693.02 
                 1193.785 
                 181327.5 
                 0 
                 6499.56 
                 106070.3 
                 0 
                 0 
               
               
                 Onion + A 
                 2483.225 
                 3120.72 
                 210744.8 
                 80495.75 
                 0 
                 2018109 
                 1178889 
                 662581.3 
               
               
                 Onion + C 
                 2315.185 
                 3150.62 
                 230510.4 
                 0 
                 7508.335 
                 66388.54 
                 0 
                 0 
               
               
                 Onion + M 
                 4465.355 
                 5859.28 
                 174097.2 
                 0 
                 6775.39 
                 65224.56 
                 0 
                 0 
               
               
                 Onion + O 
                 3948.38 
                 2365.075 
                 207007.1 
                 32371.53 
                 8053.19 
                 51931.13 
                 0 
                 0 
               
               
                 Onion + PA 
                 349.035 
                 0 
                 157786.8 
                 63239.89 
                 35.69 
                 778642.9 
                 823956.6 
                 635776.7 
               
               
                 Onion + PC 
                 1542.78 
                 0 
                 158048.5 
                 0 
                 6393.405 
                 135662.2 
                 0 
                 0 
               
               
                 Onion + PM 
                 3724.125 
                 2460.485 
                 242563.5 
                 0 
                 7737.605 
                 64012.97 
                 0 
                 0 
               
               
                 Onion + PO 
                 2927.975 
                 0 
                 4022.17 
                 103002.6 
                 8906.34 
                 28101.33 
                 0 
                 0 
               
               
                 Onion + AC 
                 870.985 
                 172.855 
                 154132.9 
                 3270.245 
                 0 
                 1352188 
                 703896.7 
                 168898.5 
               
               
                 Onion + AM 
                 3819.485 
                 4364.075 
                 397188.6 
                 0 
                 8886.73 
                 295456.1 
                 0 
                 0 
               
               
                 Onion + AO 
                 5315.445 
                 2738.63 
                 804135.2 
                 0 
                 12534.94 
                 110744.7 
                 0 
                 0 
               
               
                 Onion + CM 
                 2316.125 
                 2367.36 
                 53721.73 
                 21580.93 
                 6798.895 
                 54605.38 
                 0 
                 0 
               
               
                 Onion + CO 
                 4618.54 
                 4965.92 
                 325989 
                 0 
                 8858.455 
                 57258.25 
                 0 
                 0 
               
               
                 Onion + MO 
                 2593.645 
                 1208.055 
                 47142.54 
                 30189.94 
                 6981.475 
                 46294.3 
                 0 
                 0 
               
               
                 Onion + PAC 
                 1074.25 
                 1319.825 
                 180493.4 
                 60880.81 
                 839.535 
                 1523485 
                 2044270 
                 0 
               
               
                 Onion + PCM 
                 2273.875 
                 2775.54 
                 130204.8 
                 8038.085 
                 6408.705 
                 60792.13 
                 0 
                 0 
               
               
                 Onion + PMO 
                 2665.625 
                 0 
                 3630.715 
                 185626.6 
                 0 
                 0 
                 0 
                 0 
               
               
                 Onion + ACM 
                 2119.25 
                 8060.89 
                 452157.8 
                 0 
                 9219.665 
                 239434.4 
                 0 
                 0 
               
               
                 Onion + ACO 
                 2657.61 
                 91.855 
                 382264.7 
                 3807.255 
                 5657.73 
                 271087.9 
                 84021.03 
                 178077.2 
               
               
                 Onion + CMO 
                 2052.11 
                 4831.3 
                 193067.1 
                 40816.52 
                 8659.375 
                 57585.38 
                 0 
                 0 
               
               
                 Onion + PAM 
                 3831.155 
                 4976.68 
                 407922.3 
                 0 
                 8710.7 
                 395173 
                 0 
                 0 
               
               
                 Onion + PAO 
                 2883.325 
                 0 
                 4096.515 
                 179563.2 
                 0 
                 0 
                 0 
                 0 
               
               
                 Onion + AMO 
                 3574.2 
                 0 
                 6023.205 
                 128740.8 
                 4125.775 
                 17038.27 
                 0 
                 0 
               
               
                 Onion + PACM 
                 3995.13 
                 5006.945 
                 438052.3 
                 0 
                 8743.66 
                 435610.8 
                 0 
                 0 
               
               
                 Onion + ACMO 
                 4571.91 
                 0 
                 16464.78 
                 95371.73 
                 13783.19 
                 18421.19 
                 0 
                 0 
               
               
                 Onion + PCMO 
                 2316.075 
                 0 
                 3227.795 
                 162944 
                 0 
                 0 
                 0 
                 0 
               
               
                 Onion + PAMO 
                 2640.74 
                 0 
                 3329.67 
                 156026.2 
                 9407.52 
                 0 
                 0 
                 0 
               
               
                 Onion + PACO 
                 2493.555 
                 0 
                 3315.165 
                 166471.1 
                 0 
                 0 
                 0 
                 0 
               
               
                 Onion + 
                 2250.215 
                 0 
                 2869.585 
                 135063.8 
                 16208.05 
                 0 
                 0 
                 0 
               
               
                 PACMO 
               
               
                   
               
               
                   
                 22.12-1- 
                 22.68-1- 
                 24.50-1- 
                 26.61-1- 
                 30.40-1- 
                 31.12-1- 
                 31.77-1- 
                 36.57-1- 
               
               
                   
                 A 
                 A 
                 A 
                 A 
                 A 
                 A 
                 A 
                 A 
               
               
                   
               
               
                 Onion + P 
                 0 
                 2426.355 
                 113624.9 
                 0 
                 0 
                 132.435 
                 1080.475 
                 7066.74 
               
               
                 Onion + A 
                 0 
                 0 
                 216312 
                 0 
                 1774.785 
                 0 
                 19254.47 
                 12528.74 
               
               
                 Onion + C 
                 0 
                 4501.415 
                 116601.5 
                 0 
                 0 
                 0 
                 1057.585 
                 6373.695 
               
               
                 Onion + M 
                 0 
                 6150.535 
                 159333.6 
                 0 
                 0 
                 0 
                 858.7 
                 5750.87 
               
               
                 Onion + O 
                 0 
                 0 
                 45718.45 
                 12541.3 
                 240.335 
                 0 
                 293.375 
                 7818.715 
               
               
                 Onion + PA 
                 0 
                 0 
                 131305.2 
                 0 
                 7681.48 
                 0 
                 982.085 
                 5511.465 
               
               
                 Onion + PC 
                 0 
                 0 
                 106519.6 
                 0 
                 0 
                 0 
                 1913.52 
                 6595.775 
               
               
                 Onion + PM 
                 0 
                 7203.135 
                 166396.8 
                 0 
                 0 
                 0 
                 3179.39 
                 6564.6 
               
               
                 Onion + PO 
                 0 
                 0 
                 14555.21 
                 0 
                 335.925 
                 0 
                 394.945 
                 5312.25 
               
               
                 Onion + AC 
                 0 
                 0 
                 122263.7 
                 0 
                 0 
                 0 
                 1907.135 
                 4484.05 
               
               
                 Onion + AM 
                 0 
                 0 
                 194972.2 
                 0 
                 0 
                 0 
                 11027.67 
                 8133.615 
               
               
                 Onion + AO 
                 0 
                 0 
                 79399.2 
                 0 
                 654.82 
                 0 
                 12767.88 
                 7872.27 
               
               
                 Onion + CM 
                 0 
                 5381.76 
                 142288 
                 0 
                 352.105 
                 0 
                 2694.5 
                 5871.85 
               
               
                 Onion + CO 
                 0 
                 1726.79 
                 56373.3 
                 10865.8 
                 834.765 
                 0 
                 541.645 
                 6969.6 
               
               
                 Onion + MO 
                 5179.31 
                 0 
                 86884.65 
                 0 
                 0 
                 0 
                 1395.765 
                 5802.545 
               
               
                 Onion + PAC 
                 0 
                 0 
                 198688.5 
                 0 
                 0 
                 0 
                 16370.36 
                 9354.28 
               
               
                 Onion + PCM 
                 0 
                 6416.12 
                 166730.2 
                 0 
                 515.51 
                 0 
                 1971.4 
                 5263.63 
               
               
                 Onion + PMO 
                 1003.565 
                 656.39 
                 6990.07 
                 0 
                 596.415 
                 0 
                 594.54 
                 6361.13 
               
               
                 Onion + ACM 
                 0 
                 4902.485 
                 201632.1 
                 0 
                 0 
                 0 
                 9960.2 
                 8323.95 
               
               
                 Onion + ACO 
                 0 
                 0 
                 54839.82 
                 0 
                 660.13 
                 0 
                 11175.01 
                 6815.635 
               
               
                 Onion + CMO 
                 0 
                 1621.71 
                 94525.36 
                 0 
                 0 
                 0 
                 2180.31 
                 5368.785 
               
               
                 Onion + PAM 
                 0 
                 4212.355 
                 198890.5 
                 0 
                 0 
                 0 
                 10347.87 
                 6872.2 
               
               
                 Onion + PAO 
                 0 
                 1512.335 
                 5254.165 
                 0 
                 773.325 
                 0 
                 1234.345 
                 5189.76 
               
               
                 Onion + AMO 
                 0 
                 661.495 
                 31290.68 
                 0 
                 929.74 
                 0 
                 1204.225 
                 5361.625 
               
               
                 Onion + PACM 
                 0 
                 5327.47 
                 211331.6 
                 0 
                 0 
                 0 
                 12797.71 
                 6871.59 
               
               
                 Onion + ACMO 
                 0 
                 2663.57 
                 39258.62 
                 0 
                 253.485 
                 0 
                 825.25 
                 5063.71 
               
               
                 Onion + PCMO 
                 202.345 
                 817.015 
                 6254.42 
                 0 
                 653.04 
                 0 
                 389.37 
                 5342.37 
               
               
                 Onion + PAMO 
                 0 
                 1511.025 
                 5527.71 
                 0 
                 725.395 
                 0 
                 495.87 
                 6155.415 
               
               
                 Onion + PACO 
                 371.805 
                 1415.84 
                 5841.365 
                 0 
                 1170.295 
                 0 
                 993.62 
                 5381.35 
               
               
                 Onion + 
                 0 
                 762.83 
                 4277.475 
                 0 
                 700.005 
                 0 
                 430.555 
                 5493.65 
               
               
                 PACMO 
               
               
                   
               
               
                   
                 39.86-1- 
                 45.99-1- 
                 47.42-1- 
                 50.43-1- 
                 60.42-1- 
                 62.30-1- 
                 65.22-1- 
                 83.83-1- 
               
               
                   
                 A 
                 A 
                 A 
                 A 
                 A 
                 A 
                 A 
                 A 
               
               
                   
               
               
                 Onion + P 
                 791.44 
                 1971.66 
                 12461.52 
                 0 
                 0 
                 14409.05 
                 4932.85 
                 37604.29 
               
               
                 Onion + A 
                 4254.635 
                 7862.125 
                 11425.15 
                 0 
                 4134.66 
                 17076.94 
                 4827.97 
                 33470.48 
               
               
                 Onion + C 
                 579.955 
                 3142.855 
                 12526.67 
                 0 
                 0 
                 14532.09 
                 5130 
                 37199.48 
               
               
                 Onion + M 
                 301.52 
                 2253.135 
                 3931.64 
                 0 
                 0 
                 14500.55 
                 5319 
                 18560.28 
               
               
                 Onion + O 
                 189.005 
                 15974.93 
                 5655.435 
                 3694.96 
                 0 
                 14914.19 
                 7885.055 
                 18462.4 
               
               
                 Onion + PA 
                 407.095 
                 5864.46 
                 13115.24 
                 0 
                 2896.865 
                 16666.41 
                 4400.805 
                 33203.46 
               
               
                 Onion + PC 
                 450.035 
                 2490.64 
                 14312.61 
                 0 
                 0 
                 15746.29 
                 4649.505 
                 37919.34 
               
               
                 Onion + PM 
                 868.78 
                 1743.625 
                 1757.73 
                 155.245 
                 0 
                 15374.86 
                 4338.465 
                 12640.75 
               
               
                 Onion + PO 
                 47.65 
                 865.67 
                 0 
                 1466.865 
                 0 
                 16353.05 
                 15705.96 
                 3055.185 
               
               
                 Onion + AC 
                 686.21 
                 5756.285 
                 11270.95 
                 0 
                 3360.845 
                 17181.48 
                 4233.89 
                 34483.77 
               
               
                 Onion + AM 
                 1336.845 
                 4312.795 
                 3149.005 
                 0 
                 0 
                 16924.15 
                 4711.195 
                 13159.3 
               
               
                 Onion + AO 
                 1128.445 
                 12013.47 
                 3374.255 
                 0 
                 0 
                 15846.81 
                 4014.145 
                 14955.92 
               
               
                 Onion + CM 
                 344.505 
                 2731.66 
                 2709.225 
                 12857.02 
                 0 
                 15855.5 
                 3441.355 
                 13629.57 
               
               
                 Onion + CO 
                 252.63 
                 16628.3 
                 7944.205 
                 0 
                 0 
                 15305.37 
                 3998.115 
                 24583.82 
               
               
                 Onion + MO 
                 404.575 
                 1356.07 
                 380.035 
                 2891.2 
                 0 
                 16180.55 
                 4608.655 
                 5809.57 
               
               
                 Onion + PAC 
                 1362.035 
                 6647.695 
                 9668.645 
                 0 
                 3057.505 
                 17841.94 
                 3860.065 
                 31760.76 
               
               
                 Onion + PCM 
                 328.28 
                 2354.36 
                 2704.65 
                 0 
                 0 
                 15337.36 
                 4125.76 
                 14868.14 
               
               
                 Onion + PMO 
                 220.17 
                 1697.535 
                 0 
                 0 
                 0 
                 16063.16 
                 15129.99 
                 2271.765 
               
               
                 Onion + ACM 
                 1946.56 
                 5809.655 
                 3451.585 
                 0 
                 0 
                 15711.53 
                 4117.495 
                 14112.96 
               
               
                 Onion + ACO 
                 2156.645 
                 16052.89 
                 5475.545 
                 0 
                 714.915 
                 16314.44 
                 3686.255 
                 17859.47 
               
               
                 Onion + CMO 
                 519.205 
                 7024.215 
                 1274.395 
                 2958.735 
                 0 
                 15513.42 
                 4599.02 
                 7821.2 
               
               
                 Onion + PAM 
                 924.495 
                 4010.56 
                 2975.07 
                 0 
                 0 
                 15476.17 
                 3876.755 
                 13543.89 
               
               
                 Onion + PAO 
                 125.8 
                 3153.325 
                 0 
                 0 
                 0 
                 15508.17 
                 14357.31 
                 1861.34 
               
               
                 Onion + AMO 
                 0 
                 4687.68 
                 0 
                 18573.64 
                 0 
                 16198.68 
                 7521.145 
                 2509 
               
               
                 Onion + PACM 
                 1178.375 
                 7910.8 
                 3860.415 
                 0 
                 0 
                 16091.45 
                 3735.945 
                 14666.91 
               
               
                 Onion + ACMO 
                 218.745 
                 8066.205 
                 0 
                 9098.36 
                 0 
                 15202.36 
                 7858.95 
                 2495.935 
               
               
                 Onion + PCMO 
                 0 
                 3626.405 
                 0 
                 0 
                 0 
                 15192.24 
                 13615.49 
                 1975.02 
               
               
                 Onion + PAMO 
                 135.815 
                 4319.845 
                 0 
                 0 
                 0 
                 16372.64 
                 14199.29 
                 1812.8 
               
               
                 Onion + PACO 
                 205.34 
                 2949.905 
                 0 
                 0 
                 0 
                 15904.59 
                 14038.02 
                 1786.025 
               
               
                 Onion + 
                 0 
                 4234.15 
                 0 
                 0 
                 0 
                 16068.85 
                 13267.05 
                 1717.265 
               
               
                 PACMO 
               
               
                   
               
               
                   
                 109.41- 
                 112.13- 
                 114.86- 
                 130.36- 
                 139.63- 
                 159.67- 
                 193.05- 
                 109.41- 
               
               
                   
                 1-A 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
                 1-A 
               
               
                   
               
               
                 Onion + P 
                 72104.44 
                 15419.15 
                 9126.405 
                 3903.805 
                 22092.73 
                 7774.455 
                 4051.55 
                 72104.44 
               
               
                 Onion + A 
                 66300.83 
                 13754.07 
                 12516.15 
                 6243.98 
                 25107.29 
                 7920.03 
                 4495.255 
                 66300.83 
               
               
                 Onion + C 
                 65411.54 
                 13785.93 
                 9279.195 
                 3635.735 
                 26340.99 
                 8022.48 
                 4376.08 
                 65411.54 
               
               
                 Onion + M 
                 52806.61 
                 7689.305 
                 7938.865 
                 4595.055 
                 20160.5 
                 7857.31 
                 3889.59 
                 52806.61 
               
               
                 Onion + O 
                 49932.22 
                 9058.72 
                 6691.675 
                 1905.96 
                 16616.68 
                 6689.16 
                 3241.005 
                 49932.22 
               
               
                 Onion + PA 
                 48120.5 
                 12096.23 
                 11334.61 
                 3942.08 
                 21470.44 
                 8822.595 
                 3848.88 
                 48120.5 
               
               
                 Onion + PC 
                 48721.09 
                 12914.68 
                 9046.92 
                 3958.5 
                 22404.13 
                 7786.355 
                 4099.32 
                 48721.09 
               
               
                 Onion + PM 
                 38128.52 
                 4335.63 
                 6939.555 
                 3297.08 
                 15029.6 
                 7833.605 
                 2697.615 
                 38128.52 
               
               
                 Onion + PO 
                 18268.86 
                 7035.55 
                 3524.265 
                 2042.435 
                 9809.04 
                 7270.185 
                 3712.215 
                 18268.86 
               
               
                 Onion + AC 
                 52441.94 
                 12089.38 
                 12254.52 
                 5976.2 
                 25308.87 
                 8711.55 
                 3919.815 
                 52441.94 
               
               
                 Onion + AM 
                 36707.5 
                 4234.54 
                 8645.395 
                 12784.92 
                 15656.68 
                 8050.815 
                 3019.155 
                 36707.5 
               
               
                 Onion + AO 
                 44819.35 
                 5240.23 
                 8214.58 
                 9031.915 
                 15543.65 
                 7507.76 
                 4025.16 
                 44819.35 
               
               
                 Onion + CM 
                 40212.86 
                 6365.05 
                 5919.275 
                 3412.14 
                 25734.76 
                 7444.555 
                 4036.26 
                 40212.86 
               
               
                 Onion + CO 
                 48386.32 
                 7410.85 
                 8822.905 
                 2431.295 
                 16839.58 
                 7793.2 
                 4540.265 
                 48386.32 
               
               
                 Onion + MO 
                 31985.89 
                 8717.44 
                 5911.96 
                 2840.48 
                 15573.19 
                 7844.89 
                 4440.74 
                 31985.89 
               
               
                 Onion + PAC 
                 44492.68 
                 9995.685 
                 11372.47 
                 8544.435 
                 21799.75 
                 8484.41 
                 4131.44 
                 44492.68 
               
               
                 Onion + PCM 
                 39161.15 
                 4764.455 
                 7189.495 
                 4870.94 
                 14805.26 
                 8202.535 
                 3798.82 
                 39161.15 
               
               
                 Onion + PMO 
                 14918.1 
                 5964.915 
                 3239.705 
                 4651.11 
                 13363.97 
                 7378.16 
                 3650.605 
                 14918.1 
               
               
                 Onion + ACM 
                 35693.57 
                 3293.5 
                 7612.73 
                 6104.505 
                 14790.53 
                 8478.1 
                 3166.73 
                 35693.57 
               
               
                 Onion + ACO 
                 40577.55 
                 6358.445 
                 10167.19 
                 4802.155 
                 15886.1 
                 7860.245 
                 3914.765 
                 40577.55 
               
               
                 Onion + CMO 
                 29944.3 
                 7475.86 
                 5960.88 
                 2661.22 
                 15345.11 
                 7595.06 
                 3912.565 
                 29944.3 
               
               
                 Onion + PAM 
                 33568.13 
                 2674.18 
                 8107.99 
                 3894.155 
                 14635.48 
                 7867.58 
                 3907.615 
                 33568.13 
               
               
                 Onion + PAO 
                 12225.37 
                 4857.56 
                 2980.125 
                 8977.835 
                 10594.3 
                 7274.35 
                 3384.945 
                 12225.37 
               
               
                 Onion + AMO 
                 19344.58 
                 7074.21 
                 3808.56 
                 21563.79 
                 18519.15 
                 8471.01 
                 4306.885 
                 19344.58 
               
               
                 Onion + PACM 
                 33790.27 
                 2529.135 
                 8672.405 
                 9482.605 
                 14444.75 
                 7463.55 
                 3095.175 
                 33790.27 
               
               
                 Onion + ACMO 
                 17532.72 
                 5610.975 
                 4553.295 
                 11722.8 
                 12846.7 
                 7519.5 
                 3403.62 
                 17532.72 
               
               
                 Onion + PCMO 
                 12887.23 
                 4863.695 
                 3312.18 
                 3829.265 
                 12278.65 
                 8501.28 
                 3991.87 
                 12887.23 
               
               
                 Onion + PAMO 
                 12631.78 
                 4443.66 
                 3189.335 
                 17530.01 
                 12705.32 
                 7585.415 
                 3622.585 
                 12631.78 
               
               
                 Onion + PACO 
                 11026.31 
                 4360.555 
                 2817.38 
                 10079.8 
                 10759.32 
                 7098.595 
                 3681.155 
                 11026.31 
               
               
                 Onion + 
                 12047.41 
                 4070.63 
                 3161.265 
                 12182.35 
                 11277.06 
                 8118.06 
                 3496.99 
                 12047.41 
               
               
                 PACMO