MX2008006861A - Acyl transferase useful for decontamination - Google Patents

Abstract

The present invention provides an enzyme system that efficiently generates peracetic acid for use in decontamination applications. In preferred embodiments, the present invention provides a system that comprises an ester substrate, a hydrogen peroxide, and at least one acyl transferase. In some particularly preferred embodiments, the system further comprises at least one surfactant. In alternatively preferred embodiments, the present invention provides at least one wild-type and/or variant acyl transferase. The present invention finds particular use in decontamination involving a wide variety of chemical and biological warfare materials, as well as for general surface cleaning and decontamination.

Description

ACIL TRANSFERASA EMPLOYEE FOR DECONTAMINATION FIELD OF THE INVENTION The present invention provides an enzymatic system that efficiently generates peracetic acid for use in decontamination applications. In preferred embodiments, the present invention provides a system comprising an ester substrate, a hydrogen peroxide, and at least one acyl transferase. In some particularly preferred embodiments, the system further comprises at least one surfactant. In alternately preferred embodiments, the present invention provides at least one acyl transferase of the wild type and / or variant. The present invention finds particular use in decontamination involving a wide variety of chemical and biological weapon materials, as well as for general surface cleaning and decontamination.

BACKGROUND OF THE INVENTION Peracetic acid is widely accepted as a decontamination / disinfection agent. However, it is a chemical and carries with it all the problems associated with the use of chemical reagents. First, it degrades over time and at high temperatures. In addition, for large surface area cleaning / decontamination, large volumes of REF are required. : 193170 liquid chemicals. In addition, it can not be easily transported due to its corrosive action on tanker trucks. In addition, it has a large chemical footprint. That is, what is needed is a peracetic acid generation system that solves these storage and transport emissions, is active over a wide range of temperatures, and has a small chemical footprint.

BRIEF DESCRIPTION OF THE INVENTION The present invention provides an enzymatic system that efficiently generates peracetic acid in aqueous solution for use in decontamination applications. In preferred embodiments, the present invention provides a system comprising an ester substrate, a hydrogen peroxide and at least one acyl transferase. However, it is not intended that the present invention be limited to peracetic acid, such as any perishable (eg, pernonanoic acid, as well as percents made of C10-C18 long chain fatty acids or longer chains) finds use in the present invention. In addition, percents made of short chain fatty acids find use in the present invention. However, a variety of perishes find use in the present invention. In some embodiments, the present invention provides an enzyme system with an additional enzyme that forms hydrogen peroxide. In some additional embodiments, the present invention provides enzymatic systems that contain additional compounds that generate hydrogen peroxide including, but not limited to, such compounds as sodium percarbonate, glucose oxidase, urea and several others, including but not limited to those described in United States Patent Publication Series No. 10 / 581,014. In some preferred embodiments, the ester substrate is a stable alcohol ester, although it is not intended that the present invention be limited to any particular substrate. In some particularly preferred embodiments, the present invention provides a system for enzyme-assisted perhydrolysis in aqueous solution (eg, more than about 90% water, although the present invention is not intended to be limited to any particular percentage of water), which comprises at least one ester and at least one peroxide. However, it is contemplated that the present invention will find use in various aqueous systems, including those having a large percentage of water (eg, more than about 85%, more than about 95% or more than about 95% water) , as well as those with lower percentages of water (for example, less than about 85%). In some additional particularly preferred embodiments, the system further comprises at least one surfactant. In this way, in some embodiments, the system comprises at least one enzyme, at least one source of hydrogen peroxide, and at least one ester substrate in a buffer. In some additional embodiments, the system also comprises at least one detergent, while still in additional embodiments, the system also comprises at least one surfactant. Thus, various formulations are contemplated to find use in the present invention. In addition, in some embodiments, the present formulations are pH neutral, but in some particularly preferred embodiments, the enzyme systems also function in alkaline and slightly acidic environments (e.g., pH from about 6 to about 10). It is contemplated that the enzyme system of the present invention will find use in various forms, including liquids, granules, foams, emulsions, etc., designed to suit the near need. However, it is not intended that the present invention be limited to any particular format. In still further embodiments, the acyl transferase system of the present invention is used in conjunction with additional enzymes, including but not limited to proteases, amylases, cellulases, etc. In alternately preferred embodiments, the present invention provides at least one acyl transferase of the wild type and / or variant. In some preferred embodiments, the enzymes also have lipase activity. The present invention finds particular use in decontamination involving a wide variety of chemical and biological weapon materials, as well as for general surface cleaning and decontamination. In some embodiments, the present invention finds use in decontaminating materials contaminated by various toxic and / or pathogenic entities, including but not limited to toxic chemicals, mustard, VX, spores of B. an thra cis, Y. pestis, F ture lensis, fungi and toxins (eg, botulinum toxin, ricin, mycotoxins, etc.), as well as cells infected with infectious virions (eg, flavivirus, orthomyxovirus, paramyxovirus, arenavirus, rhabdovirus, arbovirus, enterovirus, bunyavirus, etc.). In some particularly preferred embodiments, the present invention provides a system that is capable of operating over a wide temperature range (e.g., from about 16 ° C to about 60 ° C). In still further embodiments, the system provides a small chemical footprint and is stable during short and / or prolonged storage. However, it is intended that the system of the present invention find use in numerous applications. In still further embodiments, the present invention finds use in decontaminating food and / or food products, including but not limited to, vegetables, fruits and / or other food items and / or food products. However, it is contemplated that the present invention will find use in the cleaning of surfaces of fruits, vegetables, eggs, meats, etc. However, it is intended that the present invention will find use in the food and / or foodstuff industries to remove contaminants from various food items and / or food products. In some particularly preferred embodiments, methods for the decomposition of food and / or food products exposed by the Food and Drug Administration and / or other food safety entities, as is known to those of skill in the art, find use with the present invention. As indicated herein, the present invention provides enzymatic systems for generation of perished in aqueous solution, suitable for use in decontamination. In some embodiments, the system comprises at least one ester substrate, at least one source of hydrogen peroxide and at least one acyl transferase enzyme. In some preferred modalities, the perished is selected from peracetic acid, pernonanoic acid, perproprionic, perbutanoic, perpentanoic, perhexanoic acid, percents made of long chain fatty acids, and percents made of short chain fatty acids. In some alternative preferred embodiments, the system further comprises at least one chemical hydrogen peroxide generation system, wherein the chemical hydrogen peroxide generation system comprises at least one chemical selected from sodium percarbonate, perborate and hydrogen peroxide. urea. In some embodiments, the system further comprises at least one enzyme hydrogen peroxide generation system selected from oxidases and their corresponding substrates. In some additional preferred embodiments, the system further comprises at least one enzyme hydrogen peroxide generating system, wherein the enzyme hydrogen peroxide generation system comprises at least one enzyme selected from glucose oxidase, sorbitol oxidase, hexose oxidase. , choline oxidase, alcohol oxidase, glycerol oxidase, cholesterol oxidase, pyranose oxidase, carboxyalcohol oxidase, L-amino acid oxidase, glycine oxidase, pyruvate oxidase, glutamate oxidase, sarcosine oxidase, lysine oxidase, lactate oxidase, vanillil oxidase, glycolate oxidase, galactose oxidase, uricase, oxalate oxidase, xanthine oxidase, and wherein the enzyme hydrogen peroxide generation system further comprises at least one suitable substrate for at least one enzyme. In some still further embodiments, the system further comprises at least one additional enzyme. In some preferred embodiments, at least one additional enzyme is selected from proteases, cellulases, amylases and enzymes that degrade the microbial cell wall. In some additional embodiments, at least one ester substrate is an alcohol ester. In some additional embodiments, the system further comprises at least one surfactant. In some preferred embodiments, the system further comprises at least one detergent. In some additional embodiments, the system is in a selected form of liquids, granules, foams and emulsions. The present invention also provides methods for decontamination, comprising the steps of: providing an article in need of decontamination, and at least one system for generation of perished in aqueous solution, suitable for use in decontamination; and expose the item to the system under conditions such that the item is decontaminated. In some embodiments, the exposure comprises exposing the article to the system under alkaline or acid pH conditions. In some alternate embodiments, the exposure comprises exposing the article to the system under neutral pH conditions. In some still further embodiments, the exposure comprises exposing the article to high temperature. In some preferred embodiments, the high temperature is about 60 ° C or higher. However, it is not intended that the present invention be limited to any particular temperature, as various temperatures find use in the methods of the present invention. In some embodiments, the system is in a selected form of liquids, granules, foams and emulsions. In some still further preferred embodiments, the system comprises at least one ester substrate, at least one source of hydrogen peroxide, and at least one acyl transferase. In some particularly preferred embodiments, the permeate is selected from peracetic acid, pernonanoic acid, perproprionic, perbutanoic, perpentanoic, perhexanoic acid, percents made from long chain fatty acids, and percents made from short chain fatty acids. In some alternative preferred embodiments, the method further comprises at least one chemical hydrogen peroxide generation system selected from percarbonate, perborate and hydrogen urea peroxide. In some additional alternative embodiments, the method further comprises at least one enzyme hydrogen peroxide generation system, selected from oxidases and their corresponding substrates. In some particularly preferred embodiments, the system comprises at least one enzyme hydrogen peroxide generation system, selected from glucose oxidase, sorbitol oxidase, hexose oxidase, choline oxidase, alcohol oxidase, glycerol oxidase, cholesterol oxidase, pyranose oxidase, carboxyalcohol oxidase , L-amino acid oxidase, glycine oxidase, pyruvate oxidase, glutamate oxidase, sarcosine oxidase, lysine oxidase, lactate oxidase, vanillil oxidase, glycolate oxidase, galactose oxidase, uricase, oxalate oxidase, xanthine oxidase, and where the generation system of Enzymatic hydrogen peroxide further comprises at least one suitable substrate for at least one enzyme. In additional embodiments, the method further comprises at least one enzyme or at least one additional enzyme. In some preferred embodiments, at least one enzyme is selected from proteases, amylases, cellulases and enzymes that degrade the microbial cell wall. In some alternative embodiments, at least one ester substrate is an ester alcohol. In some additional embodiments, the method further comprises at least one surfactant. In some preferred embodiments, the decontamination comprises decontaminated articles contaminated by at least one toxin and / or at least one pathogen. In some preferred embodiments, the toxin is selected from toxin butyline, anthrax toxin, ricin, scombroid toxin, ciguatoxin, tetrodotoxin and mycotoxins. In additional preferred embodiments, the pathogen is selected from bacteria, viruses, fungi, parasites and prions. In some particularly preferred embodiments, at least one pathogen is selected from Bacillus spp. , B. anthracis, Clostridium um spp. , C. botulinum, C. perfringens, Listeria spp. , Staphylococcus spp. , Streptococcus spp. , Salmonella spp. , Shigella ssp. , E. coli, Yersinia spp. , Y. pestis, Francisella spp. , F. tularensis, Camplyobacter ssp. , Vibrio spp. , Brucella spp. , Cryptosporidium spp. , Giardia spp. , Cyclospora spp. , and Trichinella spp. In still preferred embodiments, the article in need of decontamination is selected from hard surfaces, fabrics, foodstuffs, foodstuffs, clothing, rugs, carpets, textiles, medical instruments and veterinary instruments. In some particularly preferred embodiments, the food is selected from fruits, vegetables, fish, seafood, and meat. In some still further preferred embodiments, the hard surfaces are selected from domestic surfaces and industrial surfaces. In some particularly preferred embodiments, domestic surfaces are selected from kitchen countertops, sinks, cabinets, chopping boards, tables, shelves, food preparation storage areas, bathroom appliances, floors, ceilings, walls and bedroom areas. In some particularly preferred alternative modes, industrial surfaces are selected from food processing areas, food processing areas, tables, shelves, floors, ceilings, walls, sinks, chopping tables, airplanes, automobiles, trains and boats. The present invention also provides methods for decontamination comprising the steps of: providing an article in need of decontamination, and at least one system for generation of perished in aqueous solution, suitable for use in decontamination; generate the peracid in aqueous solution; and exposing the article to the perished in aqueous solution under conditions such that the article is decontaminated. In some embodiments, the exposure comprises exposing the article to the system under alkaline or acid pH conditions. In some alternative embodiments, the exposure comprises exposing the article to the system under neutral pH conditions. In some still further embodiments, the exposure comprises exposing the article to high temperature. In some preferred embodiments, the high temperature is about 60 ° C or higher. However, it is not intended that the present invention be limited to any particular temperature, as various temperatures find use in the methods of the present invention. In some embodiments, the system is in a selected form of liquids, granules, foams and emulsions. In some still further preferred embodiments, the system comprises at least one ester substrate, at least one source of hydrogen peroxide and at least one acyl transferase. In some particularly preferred embodiments, the peracid is selected from peracetic acid, pernonanoic acid, perproprionic, perbutanoic, perpentanoic, perhexanoic acid, percents made from long chain fatty acids, and percents made from short chain fatty acids. In some alternative preferred embodiments, the method further comprises at least one chemical hydrogen peroxide generation system, selected from sodium percarbonate, perborate and hydrogen urea peroxide. In some additional alternative embodiments, the method further comprises at least one enzyme hydrogen peroxide generation system, selected from oxidases and their corresponding substrates. In some particularly preferred embodiments, the system comprises at least one enzyme hydrogen peroxide generation system selected from glucose oxidase, sorbitol oxidase, hexose oxidase, choline oxidase, alcohol oxidase, glycerol oxidase, cholesterol oxidase, pyranose oxidase, carboxyalcohol oxidase, oxidase of L-amino acid, glycine oxidase, pyruvate oxidase, glutamate oxidase, sarcosine oxidase, lysine oxidase, lactate oxidase, vanillyl oxidase, glycolate oxidase, galactose oxidase, uricase, oxalate oxidase, xanthine oxidase, and wherein the peroxide generation system of Enzymatic hydrogen further comprises at least one suitable substrate for at least one enzyme. In additional embodiments, the method further comprises at least one enzyme or at least one additional enzyme. In some preferred embodiments, at least one enzyme is selected from proteases, amylases, cellulases, and enzymes that degrade the microbial cell wall. In some alternative embodiments, at least one ester substrate is an ester alcohol. In some additional embodiments, the method further comprises at least one surfactant. In some preferred embodiments, decontamination comprises decontaminating articles contaminated by at least one toxin and / or at least one pathogen. In some preferred embodiments, the toxin is selected from botulinum toxin, anthrax toxin, ricin, scombroid toxin, ciguatoxin, tetrodotoxin, and mycotoxins. In additional preferred embodiments, the pathogen is selected from bacteria, viruses, fungi, parasites and prions. In some particularly preferred embodiments, at least one pathogen is selected from Ba cillus spp. , B. an thracis, Clostridium spp. , C. botulinum, C. perfringens, Listeria spp. , Staphylococcus spp. , Streptococcus spp. , Salmonella spp. , Shigella ssp. , E. coli, Yersinia spp. , Y. pestis, Francisella spp. , F. tularensis, Camplyobacter ssp. , Vibrio spp. , Brucella spp. , Cryptosporidi um spp. , Giardia spp. , Cyclospora spp. , and Trichinella spp. In still further preferred embodiments, the item in need of decontamination is selected from hard surfaces, fabrics, food, food products, clothing, rugs, carpets, textiles, medical instruments and veterinary instruments. In some particularly preferred embodiments, the food is selected from fruits, vegetables, fish, shellfish and meat. In some still further preferred embodiments, the hard surfaces are selected from domestic surfaces and industrial surfaces. In some particularly preferred embodiments, domestic surfaces are selected from kitchen countertops, sinks, cabinets, chopping boards, tables, shelves, food preparation storage areas, bathroom appliances, floors, ceilings, walls and bedroom areas. In some particularly preferred alternative modes, industrial surfaces are selected from food processing areas, food processing areas, tables, shelves, floors, ceilings, walls, sinks, chopping tables, airplanes, automobiles, trains and boats.

BRIEF DESCRIPTION OF THE FIGURES Figure 1 provides a graph showing the enzymatic generation of peracetic acid from peroxide or hydrogen percarbonate. Figure 2 provides a graph showing the generation of peracetic acid from glucose and propylene glycol diacetate.

Figure 3 provides a graph showing the generation of peracetic acid at three different temperatures (21 ° C, 40 ° C and 60 ° C). Figure 4 provides a graph showing the ability of the enzyme acyl transferase to produce concentrated peracetic acid.

DETAILED DESCRIPTION OF THE INVENTION The present invention provides an enzymatic system that efficiently generates peracetic acid for use in decontamination applications. In preferred embodiments, the present invention provides a system comprising an ester substrate, a hydrogen peroxide, and at least one acyl transferase. However, it is not intended that the present invention be limited to peracetic acid, such as any peracid (e.g., pernonanoic acid, as well as percents made from C10-C18 long chain fatty acids or longer chains), find use in the present invention. However, a variety of perishes find use in the present invention. In some particularly preferred embodiments, the system further comprises at least one surfactant. In alternately preferred embodiments, the present invention provides at least one acyl transferase of the wild type and / or variant. The present invention finds particular use in decontamination involving a wide variety of chemical and biological weapon materials, as well as for decontamination and general surface cleaning. The present invention provides numerous advantages over currently used methods using peracid for cleaning, disinfecting and / or decontaminating. For example, the present invention facilitates the rapid generation of perished in si tu. In addition, the careful sequential addition of ingredients, peracid extraction and enzyme removal by filtration typical of current methods, is avoided by the present invention. While direct peracetic acid (ie, undiluted), is used for spore removal, it decomposes, high levels can be corrosive, it is a biohazard, there are transmission emissions with the chemical and there are limits on storage capacities. Similarly, hydrogen peroxide is used, but suffers from similar emissions such as peracetic acid. The enzymatic decontamination compositions and methods of the present invention provide numerous significant advantages over the currently used solvent-based reactive chemistry decontaminants. Some of these advantages include the non-caustic characteristics of the aqueous-based enzyme system, which allows users to safely use the system without fear of injury to themselves, equipment or the environment. In addition, there is a reduced logistic limit, in that enzymatic systems are easily transported, easy to use, and require much less water than traditional decontamination methods. In addition, with traditional methods, there is a need to collect post-decontaminant derivatives. In contrast, the surfactants used in the present invention are biodegradable. Peracid decomposes spontaneously to acetic acid or propionic acid, both of which are also biodegradable. The generation of peracid in itself in water, which occurs with the enzymatic system of the present invention, is desirable as it is much safer than the generation of reactive chemicals in solvents and requires much less bulk. However, the enzymatic activation of hydrogen peroxide has a smaller chemical footprint, and the use of enzymatic activation can also control the lifetime of the peracid. further, since hydrogen peroxide has a poor shelf life, the use of percarbonate or an equivalent in the present system avoids this problem, in addition to avoiding shipping issues associated with hydrogen peroxide. The use of percarbonate, or another compound that generates hydrogen peroxide, instead of hydrogen peroxide, also offers flexibility in the formulation components. In some preferred embodiments, the formulations comprise ingredients that are inactive until activated by exposure to water. Therefore, these formulations are especially suitable to be adjusted to the types of materials to be decontaminated, formulation compatibility, and the use of additives (as necessary), to provide optimum effectiveness. In addition, to find use as liquids, the enzyme system of the present invention also finds use as compact and dry products, as well as gels, emulsions, etc. Thus, the present invention provides the desired flexibility of formulation design, such that the formulation chosen for use is the best for such an application.

Definitions Unless defined otherwise herein, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. For example, Singleton and Sainsbury, Dictionary of Mycrobiology and Molecular Biology, 2d Ed., John Wiley and Sons, NY (1994); and Hale and Marham, The Harper Collins Dictionary of Biology, Harper Perennial, NY (1991), provide those of skill in the art, with general dictionaries of many of the terms used in the invention. Although many methods and materials similar or equivalent to those described herein find use in the practice of the present invention, the present methods and materials are described herein. Therefore, the terms defined immediately below are more fully described by reference to the Specification as a whole. Also, as used herein, the singular terms "a", "one" and "the" include plural references unless the context otherwise indicates. Unless otherwise indicated, nucleic acids are written from left to right in the orientation 5 'to 3'; the amino acid sequences are written from left to right in amino to carboxy orientation, respectively. It is understood that this invention is not limited to the particular methodology, protocols and reagents described, as these may vary, depending on the context in which they are used by those skilled in the art. It is intended that each maximum numerical limitation given through this specification include each lower numerical limitation, as if such lower numerical limitations were expressly written here. Each minimum numerical limitation given through this specification shall include each superior numerical limitation, as if such superior numerical limitations were expressly written here. Each numerical interval given through this specification will include each narrower numerical range that falls within such larger numerical ranges, as if such narrower numerical ranges were all expressly written here. As used herein, the term "an item in need of decontamination" refers to anything that needs to be decontaminated. The article is not intended to be limited to any particular item or type of article. For example, in some modalities, the article is a hard surface, while in other modalities, the article is an article of clothing. In still further modalities, the article is a textile. In still further modalities, the article is used in the medical and / or veterinary fields. In some preferred embodiments, the article is a surgical instrument. In additional modalities, the article is used in transportation (for example, roads, runways, railroads, trains, automobiles, airplanes, boats, etc.). In additional embodiments, the term is used with reference to foods and / or food products, including but not limited to meat, meat derivatives, fish, seafood, vegetables, fruits, dairy products, grains, baking products, silages, hay, fodder, etc. However, the term is intended to encompass any article that is suitable for decontamination using the methods and compositions provided herein. As used herein, the term "decontamination" refers to the removal of contaminants from an article. In some preferred embodiments, decontamination encompasses disinfection, while in other embodiments, the term encompasses sterilization. However, the term is not intended to be limited to these modalities, as the term is proposed to encompass the removal of inanimate contaminants, as well as microbial contamination (eg, bacterial, fungal, viral, prions, etc.). How it is used here, the term "disinfect" refers to the removal of contaminants from surfaces, as well as the inhibition or elimination of microbes on the surfaces of articles. It is not intended that the present invention be limited to any particular surface, article or contaminants or microbes to be removed. As used herein, the term "sterilize" refers to the removal of all microbial organisms on a surface. As used herein, the term "sporicide" refers to the elimination of microbial spores, including but not limited to fungal and bacterial spores. The term encompasses compositions that are effective in the prevention of spore germination, as well as those compositions that provide completely non-viable spores. As used herein, the terms "bactericidal", "fungicidal" and "viricidal" refer to compositions that eliminate bacteria, fungi and viruses, respectively. The term "microbicide" refers to compositions that inhibit the growth and / or replication of any microorganism, including but not limited to bacteria, fungi, viruses, protozoa, riches, etc. As used herein, the terms "bacteriostatic", "fungistatic" and "virostatic" refer to compositions that inhibit the growth and / or replication of bacteria, fungi and viruses, respectively. The term "microbiostatic" refers to compositions that inhibit the growth and / or replication of any of the organisms, including but not limited to bacteria, fungi, viruses, protozoa, riches, etc. As used herein, the term "acyl transferase" refers to an enzyme that is capable of catalyzing a reaction that results in the formation of sufficiently high amounts of peracid suitable for application, such as cleaning, bleaching and disinfection. In particularly preferred embodiments, the acyl transferase enzymes of the present invention produce very high perhydrolysis to hydrolysis ratios. The ratios of perhydrolysis to high hydrolysis of these different enzymes makes these enzymes suitable for use in a very wide variety of applications. In further preferred embodiments, the acyl transferases of the present invention are characterized as having different tertiary structure and primary sequence. In particularly preferred embodiments, the acyl transferases of the present invention comprise different primary and tertiary structures. In some particularly preferred embodiments, the acyl transferases of the present invention comprise distinct quaternary structure. In some preferred embodiments, the acyl transferase of the present invention is the acyl transferase M. smegma tis (MsAcT), while in alternative embodiments, the acyl transferase is a variant of this acyl transferase, while in still further embodiments, the acyl transferase is a homolog of this acyl transferase. In preferred embodiments, a monomer hydrolase is engineered to produce a monomeric or multimeric enzyme having acyl transferase activity better than the original monomer. However, it is not intended that the present invention be limited to this specific acyl transferase M. smegma tis, specific variants of this acyl transferase, without specific homologs of this acyl transferase. In some particularly preferred embodiments, the acyl transferase is the wild-type M. smegma tis acyl transferase described and disclosed in WO 05/056782, incorporated herein by reference in its entirety. In some particularly preferred alternative modalities, acyl transferase is one of the variant enzymes or homologs described and disclosed in WO 05/056782. In some particularly preferred embodiments, the variant comprises the S54V substitution of MsAcT (referred to herein as the "S54V variant" or "S54V variant"). As used here, the term "multimer", refers to two or more proteins or peptides that are covalently or non-covalently associated and exist as a complex in solution. A "dimer" is a multimer that contains two proteins or peptides; a "trimer" contains three proteins or peptides, etc. As used herein, "octamer" refers to a multimer of eight proteins or peptides. As used herein, "cleaning compositions" and "cleaning formulations" refer to compositions that find use in the removal of unwanted compounds from articles to be cleaned, such as fabrics, discs, contact lenses, or other solid substrates, hair (shampoos), skin (soaps and creams), teeth (mouth rinses, toothpastes), etc. The term encompasses any of materials / compounds selected for the particular type of cleaning composition desired and the product form (e.g., liquid composition, gel, granule or atomizer), as soon as the composition is compatible with acyl transferase and other enzymes used in the composition. The specific selection of materials of limiting composition is easily made considering the surface, article or fabric to be cleaned, and the desired form of the composition for cleaning conditions during use. The terms further refer to any composition that is suitable for cleaning, bleaching, disinfecting and / or sterilizing any object and / or surface. The terms are intended to include, but are not limited to, detergent compositions (e.g., liquid detergents and / or laundry solids and fine fabric detergents; hard surface cleaning formulations, such as glass, wood, ceramic and metal coatings; and windows, carpet cleaners, oven cleaners, fabric fresheners, fabric softeners, and textile and laundry pre-observers, as well as dishwashing detergents). However, the term "cleaning composition" as used herein includes, unless otherwise indicated, granular or powder-for-all-purpose or high-strength washing agents, especially cleaning detergents; liquid washing agents, gels or in the form of paste for all purposes, especially so-called high-strength liquid (HDL) types; liquid detergents of fine fabrics; agents for manual washing of dishes or agents for washing light dishes of light resistance, especially those of the high foam type; machine dishwashing agents, including the various types of tablets, granules, liquids and rinsing aids for domestic and institutional use; disinfecting agents and liquid cleansers, including antibacterial handwashing types, cleansing bars, mouthwashes, dental cleaners, shampoos for binders or trolleys, bathroom cleaners; shampoos for hair and hair conditioners; bath gels and foams for metal baths and cleaners; as well as cleaning aids such as bleaching additives and "dyeing bars" or pre-treated types. As used herein, the terms "detergent composition" and "detergent formulation" are used with reference to mixtures which are proposed for use in a washing medium for cleaning soiled objects. In some preferred embodiments, the term is used with reference to laundry fabrics and / or garments (eg, "laundry detergents"). In alternative modes, the term refers to other detergents, such as those used to clean dishes, cutlery, etc. (for example, "dishwashing detergents"). The present invention is not intended to be limited to any particular detergent formulation or composition. However, it is not intended that in addition to acyl transferase, the term encompasses detergents containing surfactants, transferases, hydrolytic enzymes, oxide reductases, builders, bleaching agents, bleach activators, blue dye and fluorescent dyes, hardening inhibitors, masking agents, enzymatic activators, antioxidants, and solubilizers. As used herein, the term "hard surface cleaning compositions", refers to detergent compositions for cleaning hard surfaces such as floors, walls, tiles, bathroom and kitchen appliances, and the like. Such compositions are provided in any form, including but not limited to solids, liquids, emulsions, etc. As used herein, "dishwashing compositions" refer to all forms for compositions for cleaning dishes including, but not limited to, granular and liquid forms. As used herein, "fabric cleaning composition" refers to all forms of fabric cleaning detergent compositions, including but not limited to, granular, liquid and stick forms. As used herein, "textile" refers to woven fabrics, as well as staple fibers and filaments suitable for conversion or use as yarns, stitches and non-woven fabrics. The term encompasses yarns made from natural fibers, as well as synthetics (for example, manufactured). As used herein, "textile materials" is a general term for fibers, intermediate yarns, yarns, fabrics, and fabrics (for example, garments and other articles). As used herein, "fabric" encompasses any textile material. Thus, the term is intended to encompass clothing, as well as fabrics, yarns, fibers, non-woven materials, natural materials, synthetic materials and any other textile material. . As used herein, the term "compatible" means that the materials of the cleaning composition do not reduce the enzymatic activity of the acyl transferase to such an extent that the acyl transferase is not effective as desired during situations of normal use. Specific cleaning composition materials are exemplified in detail here later. As used herein, "effective amount of acyl transferase enzyme" refers to the amount of acyl transferase enzyme necessary to achieve the enzymatic activity required in the specific application (e.g., decontamination). Such effective amounts are readily assessed by one of ordinary skill in the art and are based on many factors, such as the particular enzyme variant used, the cleaning application, the specific composition of the cleaning composition, and whether a liquid or dry (for example, granular, bar), and the like. As used herein, "non-woven cleansing compositions" encompass hard surface cleaning compositions, dishwashing compositions, personal care cleansing compositions (e.g., oral cleansing compositions, dental cleansing compositions, personal cleansing compositions, etc.), and compositions suitable for use in the paper and pulp industry. As used herein, "oxidizing chemical" refers to a chemical that has the ability to bleach. The oxidizing chemical is present in a quantity, pH and temperature suitable for bleaching. The term includes, but is not limited to, hydrogen peroxide and perished. As used herein, "acyl" is the general name for organic acid groups, which are the carboxylic acid residues after the removal of the -OH group (eg, ethanoyl chloride, CH3CO-C1, is the acyl formed from ethanolic acid, CH3COO-H). The names of the individual acyl groups are formed by replacing the "ico" of the acid with "ilo". As used herein, the term "acylation" refers to the chemical transformation which replaces the acyl group (RCO-) in a molecule, generally by an active hydrogen of an -OH group. As used herein, the term "transferase" refers to an enzyme that catalyzes the transfer of functional compounds to a range of substrates. As used herein, "leaving group" refers to the nucleophile which is unfolded from the acyl donor after substitution by another nucleophile. As used herein, the term "enzymatic conversion" refers to the modification of a substrate to an intermediate or the modification of an intermediate to a final product by contacting the substrate or intermediate with an enzyme. In some modalities, the contact is made by directly exposing the substrate or intermediate to the appropriate enzyme. In other embodiments, contacting comprises exposing the substrate or intermediate to an organism that expresses and / or excretes the enzyme, and / or metabolizes the desired substrate and / or intermediary to the desired intermediate and / or final product, respectively. As used herein, the phrase "detergent stability" refers to the ability of a detergent composition. In some embodiments, the stability is assessed during the use of the detergent, while in other embodiments, the term refers to the stability of a detergent composition during storage. As used herein, the phrase "stability to proteolysis" refers to the stability of a protein (e.g., an enzyme) to support proteolysis. The term is not intended to be limited to the use of any particular protease to assess the stability of a protein. As used herein, "oxidative stability" refers to the ability of a protein to function under oxidative conditions. In particular, the term refers to the ability of a protein to function in the presence of various concentrations of H202 and / or peracid. The stability under various oxidative conditions can be measured either by standard procedures known to those in the art and / or by the methods described herein. A substantial change in oxidative stability is evidenced by at least about 5% or greater increase or reduction (in most embodiments, it is preferably an increase), in the half-life of enzyme activity, compared to the enzymatic activity present in the absence of oxidative compounds. As used herein, "pH stability" refers to the ability of a protein to function at a particular pH. In general, most enzymes have a finite pH range in which they will work. In addition, for enzymes that function at mid-range pH (i.e., approximately pH 7), there are enzymes that are capable of working under conditions with very high or very low pH. Stability at various pH can be measured either by standard procedures known to those in the art and / or by the methods described herein. A substantial change in the pH stability is evidenced by at least about 5% or greater increase or decrease (in most embodiments, it is preferably an increase) in the half-life of the enzyme activity, compared to the enzymatic activity at the optimum pH of the enzyme. However, it is not intended that the present invention be limited to any level of pH stability or pH range. As used herein, "thermal stability" refers to the ability of a protein to function at a particular temperature. In general, most enzymes have a finite range of temperatures at which they will work. In addition to enzymes that function at mid-range temperatures (eg, room temperature), there are enzymes that are capable of operating at very high or very low temperatures. The thermal stability can be measured either by known methods or by the methods described herein. A substantial range in thermal stability is evidenced by at least about 5% or greater increase or reduction (in most embodiments, it is preferably an increase), in the half-life of the catalytic activity of a mutant when exposed to a temperature different (ie, higher or lower) than the optimum temperature for enzymatic activity. However, it is not intended that the present invention be limited to any level of temperature stability or temperature range. As used herein, the term "chemical stability" refers to the stability of a protein (e.g., an enzyme) toward chemicals that adversely affect its activity. In some embodiments, such chemicals include, but are not limited to hydrogen peroxide, perishes, anionic detergents, cationic detergents, non-ionic detergents, chelators, etc. However, it is not intended that the present invention be limited to any particular level of chemical stability or chemical stability range. As used herein, the phrase "alteration in substrate specificity" refers to changes in the substrate specificity of an enzyme. In some embodiments, a change in substrate specificity is defined as a difference between the Kcat / Km ratio observed with an enzyme compared to enzymatic variants or other enzyme compositions. The specificities of enzymatic substrate vary, depending on the substrate tested. The specificity of the substrate of an enzyme is determined by comparing the catalytic efficiencies, it is exhibited with different substrates. These determinations find particular use in assessing the efficiency of mutant enzymes, as is generally desired to produce variant enzymes that exhibit higher ratios for particular substrates of interest. For example, the acyl transferase enzymes of the present invention are more efficient in producing peracid from an ester substrate than enzymes currently being used in decontamination, cleaning, bleaching and disinfection applications. Another example of the present invention is an acyl transferase with a lower activity in peracid degradation compared to the wild type. Another example of the present invention is an acyl transferase with activity in more hydrophobic acyl groups than acetic acid. However, it is not intended that the present invention be limited to any particular substrate composition, nor to any specific substrate specificity. As used herein, "surface property" is used with reference to an electrostatic charge, as well as properties such as hydrophobicity and / or hydrophilicity by the surface of a protein. As used herein, the phrase "is independently selected from the group consisting of ..." means that portions or elements that are selected from the referenced Markush group may be the same, may be different or any mixture of elements as indicated in the following example: As used herein, the terms "purified" and "isolated" refer to the removal of contaminants from a sample. For example, acyl transferases are purified by removal of contaminating proteins and other compounds within a solution or preparation that are not acyl transferases. In some embodiments, recombinant acyl transferases are expressed in fungal or bacterial host cells and these recombinant acyl transferases are purified by removal of other host cell constituents.; the percentage of recombinant acyl transferase peptides is thereby increased in the sample. As used herein, the term "derivative" refers to a protein which is derived from a protein by the addition of one or more amino acids to either or both of the C and N-terminal ends, the substitution of one or more amino acids at one or a number of different sites in the amino acid sequence, and / or deletion of one or more amino acids at either or both ends of the protein or at one or more sites in the amino acid sequence, and / or insertion of one or more more amino acids at one or more sites in the amino acid sequence. The preparation of a protein derivative is preferably achieved by modifying a DNA sequence which codes for the native protein, transformation of such a DNA sequence into a suitable host, and expression of the modified DNA sequence to form the derived protein. Related proteins (and derivatives) comprise, "variant proteins". In some preferred embodiments, variant proteins differ from a precursor protein and another by a small number of amino acid residues. The number of different amino acid residues may be one or more, preferably 1, 2, 3, 4, 5, 10, 15, 20, 30, 40, 50 or more amino acid residues. In some preferred embodiments, the number of different amino acids between variants is between 1 and 10. In some particularly preferred embodiments, related proteins and particularly variant proteins comprise at least 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98% or 99% of amino acid sequence identity. Additionally, a related protein or a variant protein as used herein, refers to a protein that differs from another related protein or a precursor protein in the number of prominent regions. For example, in some embodiments, variant proteins have 1, 2, 3, 4, 5, or 10 corresponding prominent regions that differ from the precursor protein. Several methods are known in the art that are suitable for generating variants of the acyl transferase enzymes of the present invention, including but not limited to in situ saturation mutagenesis, scanning mutagenesis, insertional mutagenesis, random mutagenesis, site-directed mutagenesis and directed evolution, as well as several other recombination procedures. In particularly preferred embodiments, homologous proteins are engineered to produce enzymes with the desired activities. In some particularly preferred embodiments, engineered proteins comprise at least one or a combination of the following conserved residues: L6, W14, W34, L38, R56, D62, L74, L78, H81, P83, M90, K97, G110 , L114, L135, F180, G205. In alternative embodiments, these engineered proteins comprise the GDSL-GRTT and / or ARTT portions. In additional embodiments, the enzymes are multimers, which include but are not limited to dimers, octamers and tetramers. In still further preferred embodiments, the engineered proteins exhibit a ratio of perhydrolysis to hydrolysis that is greater than 1. An amino acid residue of an acyl transferase is equivalent to an acyl transferase residue M. smegma tis if it is either homologous ( that is, having a corresponding position in either the primary and / or tertiary structure), or analogous to a specific residue or portion of such residue in acyl transferase M. smegma ti s (ie having the same or similar capacity functional to combine, react, and / or interact chemically). In some embodiments, to establish homology to primary structure, the amino acid sequence of an acyl transferase is directly compared to the primary sequence of acyl transferase M. smegma tis and particularly to a series of residues known to be variant in all acyl transferases for which is known the sequence. After aligning the conserved waste, allowing necessary insertions and deletions to maintain alignment (ie, avoiding the removal of conserved residues through deletion and arbitrary insertion), the residues equivalent to particular amino acids are defined in the primary acyl transferase sequence M. smegma tis. In preferred embodiments, the alignment of conserved residues retains 100% of such residues. However, alignment greater than 75% or as little as 50% of conserved waste is also adequate to define equivalent waste. In preferred embodiments, the preservation of the serine and catalytic histidine residues is maintained. Conserved residues are used to define the corresponding equivalent amino acid residues of acyl transferase M. smegma tis in other acyl transferases (eg, acyl transferases from other Mycoba cteri um species, as well as any other organisms). In some embodiments of the present invention, the DNA sequence encoding acyl transferase M. smegma tis is modified. In some embodiments, the following residues are modified: Cys7, AsplO, Serll, Leul2, Thrl3, Trpl4, TrplO, Pro24, Thr25, Leu53, Ser54, Ala55, Thr64, Asp65, Arg67, Cys77, Thr91, Asn94, Asp95, Tyr99, Vall25, Prol38, Leul40, Prol46, Prol48, Trpl49, Phel50, Ilel53, Phel54, Thrl59, Thrl86, Ilel92, Ilel94, and Phel96. However, it is not intended that the present invention be limited to the sequence that is modified in these positions. However, it is intended that the present invention encompass several modifications and combinations of modifications. In additional embodiments, equivalent residues are defined by determining the homology at the tertiary structure level for an acyl transferase whose tertiary structure has been determined by x-ray crystallography. In this context, "equivalent residues" are defined as those by which the atomic coordinates two or more of the main chain atoms of a particular amino acid residue of the carbonyl hydrolase and acyl transferase M. smegma tis (N en N, CA in CA, C in C, and 0 in 0), they are within 0.13 nm and preferably 0.1 nm after alignment. The alignment is achieved after the best model has been oriented and positioned to give the maximum overlap of atomic coordinates of hydrogen-free protein atoms of the acyl transferase in question to the acyl transferase M. smegma tis. As is known in the art, the best model is the crystallographic model given the lowest R factor for experimental diffraction data at the highest resolution available. Equivalent residues which are functionally and / or structurally analogous to a specific residue of acyl transferase M. smegma tis are defined as those amino acids of the acyl transferases which preferentially adopt a conformation so that any alter, modify or modulate the protein structure, to effect changes in substrate and / or catalyst bond in a defined manner and attributed to a specific residue of the acyl transferase M. smegma tis. In addition, they are those residues of acyl transferase (in cases where a tertiary structure has been obtained by x-ray crystallography), which occupy a position analogous to the extension through the main chain atoms of the given residue can not satisfying the equivalence criterion based on the occupation of a homologous position, the atomic coordinates at least two of the side chain atoms of the residue falling within 0.13 nm of the corresponding side chain atoms of acyl transferase M. smegma ti s . In some embodiments, some of the residues identified for substitution, insertion or deletion are conserved residues while others do not. The acyl transferase mutants of the present invention include several mutants, including those encoded by nucleic acid comprising a signal sequence. In some embodiments of acyl transferase mutants that are encoded by such a sequence are secreted by an expression host. In some additional modalities, the nucleic acid sequence comprises a homolog having a secretion signal. The characterization of wild-type mutant proteins is achieved via any suitable means and is preferably based on properties of interest. For example, pH and / or temperature, as well as detergent and / or oxidative stability are determined in some embodiments of the present invention. Indeed, it is contemplated that enzymes having various degrees of stability in one or more of these characteristics (pH, temperature, proteolytic stability, detergent stability and / or oxidative stability) will find use. In still other embodiments, acyl transferase with low peracid degradation activity is selected. As used herein, "corresponding to" refers to a residue at the position listed on a protein or peptide, or a residue that is analogous, homologous or equivalent to a residue listed on a protein or peptide. As used herein, "corresponding region" generally refers to an analogous position along related proteins or a precursor protein. The terms "encoding nucleic acid molecules", "encoding nucleic acid sequences", "encoding DNA sequences" and "encoding DNA" refer to the order or sequences of deoxyribonucleotides by a deoxyribonucleic acid strand. The order of these deoxyribonucleotides determines the order of amino acids along the polypeptide (protein) chain. The DNA sequence thus encodes the amino acid sequence. As used in this document, the term "Analogous sequence" refers to a sequence within a protein that provides similar function, tertiary structure and / or conserved residues as the protein of interest (ie, typically the original protein of interest). For example, in epitope regions containing an alpha helix or a beta sheet structure, the amino acid replaced in the analogous sequence preferably maintains the specific structure. The term also refers to nucleotide sequences, as well as amino acid sequences. In some embodiments, analogous sequences are developed in such a way that the replacement amino acids result in a variant enzyme that exhibits a similar or improved function. In some embodiments, the tertiary structure and / or conserved residue of the amino acids in the protein of interest is located in or near the segment or fragment of interest. Thus, where the segment 1 fragment of interest contains, for example, an alpha helix or a beta sheet structure, the replacement amino acids preferably maintain the specific structure. As used herein, "homologous protein" refers to a protein (e.g., acyl transferase) that has similar action and / or structure, such as a protein of interest (e.g., acyl transferase from another source). It is not intended that the homologs are necessarily evolutionarily related. In this way, the term encompassing the same or similar enzymes (ie, in terms of structure and function) obtained from different species is intended. In some preferred embodiments, it is desirable to identify a homologue having a quaternary, tertiary and / or primary structure similar to the protein of interest, replacement for the segment or fragment in the protein of interest with an analogous segment of the homolog will reduce the disorder of the change. In some embodiments, homologous proteins have induced similar immunobiological responses as a protein of interest. As used herein, "natural type" and "natural" proteins are those found in nature. The terms "wild-type sequence" and "wild-type gene" are used interchangeably herein to refer to a sequence that originates natively or naturally in a host cell.In some embodiments, the wild-type sequence refers to to a sequence of interest that is the starting point of a protein engineering design project The genes that encode the naturally occurring protein can be obtained in accordance with the general methods known to those skilled in the art. they generally comprise synthetically labeled probes that have putative sequences that encode regions of the protein of interest, prepare genomic libraries of organisms expressing the protein, and select libraries for the gene of interest by hybridization of the probes.The positively hybridizing clones are then subjected to to map and sequenced.

The term "recombinant DNA molecule" as used herein, refers to a DNA molecule comprising DNA segments joined together by molecular biological techniques. The term "recombinant oligonucleotide" refers to an oligonucleotide created using molecular biological manipulations, including but not limited to, ligation of two or more oligonucleotide sequences generated by restriction enzyme digestion of a polypeptide sequence, the synthesis of oligonucleotides (e.g., synthesis of primers or oligonucleotides) and the like. The degree of homology between sequences can be determined by any suitable method known in the art (see, for example, Smith and Waterman, Adv. Appl. Math., 2: 482 [1981]; Needleman and Wunsch, J. Mol. Biol. ., 48: 443 [1970], Pearson and Lipman, Proc. Nati, Acad. Sci. USA 85: 2444 [1988], programs such as GAP, BESTFIT, FASTA and TFASTA in Genetic Software Packages Wisconsin (Genetics Computer Group, Madison, Wl); and Deverreux et al., Nucí, Acid Res., 12: 387-395 [1984]. The phrases "substantially similar" and "substantially identical" in the context of at least two nucleic acids or polypeptides typically means that a polynucleotide or polypeptide comprises a sequence having at least about 40% identity, more preferably at least 75% identity , more preferably at least about 80% identity, still more preferably at least 90%, even more preferably about 95%, more preferably about 97% identity, sometimes as much as about 98% and about 99% sequence identity, compared to the reference sequence (ie, natural type). The sequence identity can be determined using known programs such as BLAST, ALIGN and CLUSTAL using standard parameters. (See, for example, Altschul, et al., J. Mol. Biol. 215: 403-410 [1990], Henikoff et al., Proc. Nati. Acad. Sci. USA 89: 10915 [1989]; Karin et al. , Proc. Nati, Acad Sci USA 90: 5873 [1993], and Higgins et al., Gene 73: 237-244 [1988]). The software to perform the BLAST analysis is publicly available through the National Center for Biotechnology Information. You can also search databases using FASTA (Pearson et al., Proc. Nati. Acad. Sci. USA 85: 2444-2448 [1988]). An indication that two polypeptides are substantially identical is that the first polypeptide is immunologically reacted cross-linked with the second polypeptide. Typically, polypeptides that differ by conservative amino acid substitutions are immunologically cross-reactive. Thus, a polypeptide is substantially identical to a second polypeptide, for example, wherein the two polypeptides differ only by conservative substitution. Another indication that two nucleic acid sequences are substantially identical is that the two molecules hybridize to each other under severe conditions (e.g., within a range of medium to high severity). As used herein, "equivalent residues" refers to proteins that the particular amino acid residues allow. For example, equivalent residues can be identified by determining the homology at the tertiary structure level for a protein (eg, acyl transferase) whose tertiary structure has been determined by X-ray crystallography. Equivalent residues are defined as those which the atomic coordinates of two or more of the main chain atoms of a particular amino acid residue of the protein having putative equivalent residues and the protein of interest (N in N, CA in CA, C in C and O in O) are within 0.13 nm and preferably 0.1 nm after alignment. The alignment is achieved after the best model has been oriented and placed to provide the maximum overlap of atomic coordinates of non-hydrogen protein atoms of the analyzed proteins. The preferred model is the crystallographic model providing the lower R factor for experimental diffraction data at the highest available resolution, determined using methods known to those of experience in the crystallography and protein characterization / analysis technique. The present invention provides an enzymatic system that efficiently generates para-acetic acid in water for use in decontamination applications. In preferred embodiments, the present invention provides a system comprising an ester substrate, a hydrogen peroxide, and at least one acyl transferase. However, the present invention is not intended to be limited to paracetic acid, such as any peracid (e.g., pernonic acid, as well as percents made of C10-C18 long chain fatty acids or longer chains) find use in the present invention. Of course, a variety of perished find use in the present invention. In some particularly preferred embodiments, the system further comprises at least one surfactant. In alternately preferred modalities, the present invention provides at least one acyl transferase of the wild type and / or variant. The present invention finds particular use in decontamination involving a wide variety of chemical and biological weapon materials, as well as to generate general surface cleaners and decontamination. In some embodiments, the present invention finds use in decontaminating contaminated material with materials including but not limited to toxic chemicals, mustards, VX, B spores. antrha cis, Y. pestis, F. tularensi s, fungi and toxins (eg, botulinum, ricin, mycotoxins, etc.), as well as cells infected with infectious virions (eg, flavivirus, orthomyxovirus, paramyxovirus, arenavirus, rhabdovirus , arboviruses, enteroviruses, buniaviruses, etc.). In some particularly preferred embodiments, the present invention provides systems that are capable of functionalizing at wide temperature ranges (e.g., from about 5 ° C to about 90 ° C.; from about 16 ° C to about 60 ° C; and from about 25 ° C to about 100 ° C). In still other preferred embodiments, the system provides a small chemical footprint and is stable during prolonged and / or short storage. Indeed, it is intended that the system of the present invention find use in numerous applications. It is contemplated that the enzyme system of the present invention will find use in various forms, including liquids, granules, foams, emulsions, etc., designed to adjust the near need. Indeed, it is not intended that the present invention be limited by any particular format. In still further embodiments, the acyl transferase system of the present invention was used together with additional enzymes, including but not limited to proteases, amylases, etc. Indeed, it is contemplated that various enzymes will find use in conjunction with the present invention, including but not limited to enzymes that degrade glycoprotein and that degrade the microbial cell wall, lysozymes, hemicelluloses, peroxidase, proteases, cellulases, xylanases, lipases, phospholipases, esterases, cutinases, pectinases, keratinases, reductases, oxidases, phenoloxidases, lipoxygenases, ligninases, pullulanases, tanases, pentosanas, malanases, ß-glucanases, arabinosidases, hyaluronidase, chondroitinase, laccase, endolguclanases, PNGases, amylases, etc., as well as mixtures thereof. In some embodiments, enzyme stabilizers find use in the present invention. It is contemplated that using combinations of enzymes, there will be a simultaneous reduction in the amount of chemical needs. In some particularly preferred embodiments, the present invention finds use in the enzymatic generation of percents of ether substrates and hydrogen peroxide. The present invention is not intended to be limited to any enzyme specific for the generation of hydrogen peroxide, as any enzyme that generates H2O and acid with a suitable substrate finds use in the methods of the present invention. For example, lactate oxidase from Lactobacillus species which are known to create H202 from lactic acid and oxygen, finds use with the present invention. Indeed, an advantage of the methods of the present invention is that the generation of acid reduces the pH of a basic solution to the pH range in which the peracid is most effective in bleaching (ie, at or below the pKa). Other enzymes (eg, alcohol oxidase, ethylene glycol oxidase, glycerol oxidase amino acid oxidase, etc.) which can generate hydrogen peroxide also find use with ester substrates in combination with the perhydrolase enzymes of the present invention to generate percents. Enzymes that generate acid from the substrates without the generation of hydrogen peroxidase also find use in the present invention. Examples of such enzymes include, but are not limited to, proteases. Thus, as described herein, the present invention provides definite advantages over the method and compositions currently used for decontaminant formulation and its use, as well as various other applications. In some preferred embodiments, the substrates are selected from one or more of the following: formic acid, acetic acid, propionic acid, butyl acid, valeric acid, caproic acid, caprylic acid, nonanoic acid, decanoic acid, dodecanoic acid, myristic acid, palmitic acid, stearic acid and oleic acid. In addition to the acyl transferase described herein, various hydrolases find use in the present invention, including but not limited to carboxylate ester hydrolase, thioester hydrolase, phosphate monoester hydrolase and phosphate diester hydrolase which acts on ester linkages; a thioester hydrolase which acts on ether bonds; and a-amino-acyl-peptide hydrolase, peptidyl-amino acid hydrolase, acyl-amino acid hydrolase, dipeptide hydrolase and peptidyl-peptide hydrolase which acts on peptide bonds. Such hydrolases find use alone or in combination with perhydrolases. Among the preferred ones are carboxylate ester hydrolase and peptidyl peptide hydrolase. Suitable hydrolases include: (1) proteases belonging to the peptidyl-peptide hydrolase class (e.g., pepsin, pepsin B, renin, trypsin, chymotrypsin A, chymotrypsin B, elastase, enterokinase, cathepsin C, papain, chemopapain, ficin, thrombin, fibrinolysin, renin, subtilisin, aspergilopeptidase A, collagenase, clostridiopeptidase B, kallikrein, gastrisine, cathepsin D, bromelain, keratinase, chymotrypsin C, pepsin C, aspergilopeptidase B, urokinase, carboxypeptidase A and B and aminopeptidase); (2) carboxylate ester hydrolase including carboxyl esterase, lipase, pectin esterase and chlorophyllase; and (3) enzymes having proportions of perhydrolysis at high hydrolysis. Especially effective amounts of these are lipases, as well as esterases that exhibit high proportions of perhydrolysis to high hydrolysis, as well as esterases that engineered protein, cutinase and lipase using the primary, secondary, tertiary and / or quaternary structural characteristics of the perhydrolases of the present invention. The hydrolase is incorporated into the detergent composition as required in accordance with the purpose. It will preferably be incorporated in an amount of 0.00001 to 5 weight percent, and more preferably 0.02 to 3 weight percent. This enzyme will be used in the form of granules made from crude enzyme alone or in combination with other enzymes and / or components in the detergent composition. Raw enzyme granules are used in such a way that the amount of purified enzyme is 0.001 to 50 percent. in the granules. The granules are used in an amount of 0.002 to 20 and preferably 0.1 to 10 weight percent. In some embodiments, the granules are formulated to contain an agent that protects the enzyme and a dissolving retarding material (i.e., material that regulates the dissolution of granules during use). In addition, oxidase finds use in the present invention in the present invention, which include carbohydrate oxidase selected from the group consisting of aldose oxidase (IUPAC classification EC1.1.3.9), galactose oxidase (IUPAC classification EC1.1.3.9) , cellobiose oxidase (classification EC1.1.3.25 of UIPAC), pyranose oxidase (classification ECl.1.3.10 of IUPAC), sorbose oxidase (classification EC1.1.3.11 of IUPAC) and / or hexose oxidase (classification • EC1.1.3 .5 IUPAC), glucose oxidase (EC1.1.3.4 classification of IUPAC) and mixtures thereof. Indeed, it is contemplated that any suitable oxidase that follows the equation: Enzyme + reduced substrate - > Oxidized substrate + H202 finds use in the present invention. Additional components find use in the formulations of the present invention. Although it is not intended that the formulations of the present invention be limited, several components are described herein. Indeed, while the components are not essential for purposes of the present invention, the non-limiting list of the companions illustrated hereinafter is suitable for use in the present compositions and it may be desirable to be incorporated in certain embodiments of the invention, for example to assist or improve cleaning performance, to treat the substrate to be cleaned, or to modify the aesthetic of the cleaning composition, as the case may be with perfumes, dyes, dyes and the like. It will be understood that such companions are in addition to the enzymes of the present invention, hydrogen peroxide and / or source of hydrogen peroxide, and material comprising an ester portion. The precise nature of these additional components, and levels of incorporation thereof, will depend on the physical form of the composition and the nature of the cleaning operation which will be used. Suitable companion materials include, but are not limited to, surfactants, promoters, chelating agents, agents that inhibit dye transfer, deposition aids, dispersants, corrosion inhibitors, additional enzymes and enzyme stabilizers, catalytic materials, bleach activators, bleached reinforcers, preformed permeators, polymeric dispersing agents, agents for removing soil clay / anti-redeposition, brighteners, suds suppressors, dyes, perfumes, agents for elasticizing the structure, carriers, hydrotropes, processing aids and / or pigments. In addition to the description below, suitable examples of such companions and levels of use are found in U.S. Patent Nos. 5, 576,282, 6,306,812 and 6,326,348, incorporated herein by reference. The aforementioned accompanying ingredients may constitute the balance of the cleaning compositions of the present invention. In some embodiments, the enzyme system of the present invention further comprises enzymes that remove any residual peracid and / or H202 after decontamination is achieved. Such enzymes include but are not limited to catalyzed and / or hydrolytic enzymes. Importantly, the present invention provides means for effective cleaning, bleaching and disinfection at a pH and wide temperature ranges. In some embodiments, the pH range used in this generation is 4-12. In some alternative embodiments, the temperature range used is between about 51 and about 90 ° C. The present invention provides advantages over currently used systems (See, for example, EP Application 87-304933.9) bleaching is possible at the optimum pH of peracid oxidation, as well as providing bleaching at neutral pH, acidic pH and at low temperatures.

EXPERIMENTAL The following examples are provided to further demonstrate and illustrate certain preferred embodiments and aspects of the present invention and are not construed as limiting the scope thereof. In the experimental description which is as follows, the following abbreviations apply: ° C (degrees centigrade); TA (room temperature), rpm (revolutions per minute); H20 (water); dH20 (distilled water); HCl (hydrochloric acid); aa (amino acid, pb (base pair), kb (kilobase pair), kD (kilodaltons), gm (grams), μg and ug (micrograms), mg (milligrams), ng (nanograms), μl and ul (microliters); ml (milliliters); mm (millimeters); nm (nanometers); μm and um (micrometer); M (molar); mM (millimolar); μM and uM (micromolar); U (units); V (volts); MW (molecular weight); sec (seconds); min (minutes); h (hours) MgCl2 (magnesium chloride); NaCl (sodium chloride); OD420 (optical density at 420 nm); PAGE (poliacplamide gel electrophoresis); EtOH (ethanol); LB (Luna broth); THE agar Moon); PBS (buffered phosphate salt [150 mM NaCl, mM sodium phosphate buffer, pH 7.2]); SCS (sodium dodecyl sulfate); Tris (tris (hydroxymethyl) ammomethane); p / v (weight to volume); v / v volume to volume); % p (percentage by weight); PAA (peracetic acid); Per (perhydrolase); per (perhydrolase gene); Ms (M. smegma tis); MsAcT (acyl transferase from M. smegma tis); variant 854V (vanant of acyl transferase from M. smegma tis comprising the S54V substitution); MS (mass spectroscopy); Dial (Dial Brands, Inc., Scottsdale, AZ); Kemira (Kemira Industrial Chemicals, Helsmgborg, Switzerland); EM Science (Em Science, Gibbston, NJ); HP (Hewlett-Packard, Palo Alto, CA); ICN (ICN Pharmaceuticals, Inc., Costa Mesa, CA); Dial (Dial, Corp, Scottsdale, AZ); Pierce (Pierce Ciotechnology, Rockford, IL); Amicon (Amicon, Inc., Beverly, MA); ATCC (American Type Culture Collection, Manassas, VA); Amersham (Amesham Biosciences, Inc., Piscataway, NJ); Becton Dickmson (Becton Dickmson Labware, Lincoln Park, NJ); BioRad (BioRad, Richmond, CA); Difco (Difco Laboratories, Detroit, MI); GIBCO BRL or Gibco BRL (Life Technologies, Inc., Gaithersburg, MD); MIDI (MIDI Labs, Newark, DE); Sigma or Aldrich (Sigma-Aldrich Inc., St. Louis, MO); Sorvall (Sorvall Instruments, a subsidiary of DuPont Co., Biotechnology Systems, Wilmington, DE); Agilent (Agilent Technologies, Palo Alto, CA); Minolta (Konica Minolta, Ramsey, NJ); and Zeiss (Cari Zeiss, Inc., Thornwood, NY).

EXAMPLE 1 Elimination Curve for B Spores. subtilis by Peracetic Acid (PAA) In this example, experiments are conducted to determine the peracetic acid and acetic acid removal curve together with detergent (commercially available from PUREX® [Dial] used in this Example) for B spores. subtilis. In these experiments, the spores of B. subtilis were prepared as known in the art (See, for example, Siccardi et al., J. Bacteriol., 121: 13-19 [1975]). The assays were carried out in duplicate in 96-well round-bottom microtiter plates.

(Costar) with paracetic acid (32% by weight in acetic acid; Aldrich). The PAA was serially diluted in either 50 mM buffer KP04, pH 7.1 ("Shock absorber"), or at a 1: 500 dilution of Purex (original formula; Dial) in the same buffer ("Shock absorber + Det") in a total volume of 50 μl. The amount of PAA added to the assay is 0, 0.4 or 40 mM. A volume of 5 μl of the spore suspension, containing 109-1010 spores, was then added to each well and the assay was incubated for 15 minutes at room temperature. Then ice-cooled LB (150 μl) was added (See, for example, Sambrook et al., "Molecular Cloning: A Laboratory Manual", Second Edition (Could Spring Harbor), [1989)] to each well, mixed, and 100 μl was transferred to each fresh 96-well plate. Serial dilutions of each solution are made (in a total volume of 100 μl / well). A volume of 5 μl of each dilution was stained in LA plates (Sambrook et al., Supra),, and incubated at 37 ° C for 17-24 hours.

The colonies were counted and the% of spores eliminated was determined in relation to the respective controls (buffer alone or buffer + detergent, without peracid). The results are presented in Table 1, as an average of duplicates from an experiment. However, the experiment was performed twice in duplicate with similar results. Based on these results, PAA in the range of about 4 to about 40 mM was determined to be sufficient to remove B spores. subtilis in 15 minutes.

EXAMPLE 2 Enzymatic generation of PAA In this example, three methods for generation of PAA by acyl transferase are described. In one method, at least one acyl transferase (wild type or variant) is combined with at least one ester substrate, and hydrogen peroxide in a buffer or detergent, with or without one or more surfactants. In an alternative method, at least one acyl transferase (natural type or variant), at least one ester substrate and sodium percarbonate (or other source of H202) are combined in a buffer or detergent, with or without one or more other surfactants . In yet a further method, at least one acyl transferase (wild type or variant) is combined with glucose oxidase and glucose, at a concentration sufficient to generate an amount of PAA, with which spores are removed in buffer or detergent. In some formulations, one or more other surfactants are also included. Other enzymes that H202 generates also find use in this system, which include oxidases, oxidoreductases (for example, glycerol oxidase and hexose oxidase). In some preferred embodiments, an independent alcohol oxidase co-factor was used.

Determination of PAA Concentration In these experiments, methods known in the art were used to determine the concentration of PAA (See, for example, Pinkernell et al., Analyst, 122: 567-571 [1997]). In this ABTS assay, 100 μl of the solution to be analyzed was added to 1 ml of 125 mM potassium citrate buffer, pH 5.0, containing 1.0 mM of 3-ethylbenzthiazoline-6-sulfonic acid (ABTS) and 50 μM Kl, and allowed to incubate at room temperature for 3 minutes. The absorbance at 420 nm was measured on an HP 8452A Diode Array Spectrometer and compared to a standard curve prepared using authentic standard. Enzymatic reactions to form PAA were initiated without enzyme addition and conducted at room temperature. Aliquots of the reactions were removed at the indicated times and analyzed for PAA concentration. The sodium percarbonate used in these experiments was obtained from Kemira and the hydrogen peroxide was obtained from EM Sience.

Comparison of Enzymatically Generated PAA from H202 or Sodium Percarbonate A solution of 39 mM sodium percarbonate (sodium carbonate peroxyhydrate, Technical grade 85%, providing 100 mM effective H202, Kemira) in 320 mM KP04 was prepared , pH 7.1. After dissolution of the solid percarbonate, the resulting solution has a pH of 7.6. To compare the enzymatic production of PAA of prepared H202 (32% by weight, Aldrich) or H202 formed of percarbonate under identical pH conditions, two reactions were prepared. One reaction contained 100 mM H202 in 320 mM KP04, pH 7.6, 100 mM 1,2-propylene glycol diacetate (Aldrich) and 2pp S54V variant. The absolute concentration of H202 was measured from the value declared on the label and not confirmed by analysis. The second reaction contained 39 mM sodium percarbonate in 320 mM KP04, pH 7.1, 100 mM 1,2-propylene glycol diacetate and 2 ppm S54V. The reactions were initiated by the addition of the enzyme. The samples were removed at the indicated times and the concentration of PAA was determined as described in Example 1. The results are shown in Figure 1. As indicated in this Figure, the progress of the reaction and final concentration of PAA is similar in both cases.

EXAMPLE 3 Enzymatic Generation of PAA that Eliminates Spores of B. subtilis In this example, conducted experiments are described to evaluate the elimination capacity of enzymatically generated PAA with B spores. subtilis. Based on the results obtained in the experiments described in Examples 1 and 2, a range of 4 to 40 mM PAA was determined to be sufficient to demonstrate the elimination of B spores. subtilis 1-168. In these experiments, the elimination of spore in buffer, as well as in detergent, was evaluated.

Removal of Spores in Shock Absorber In this experiment, sodium percarbonate was used as the source of H02. The final solution contained: 100 mM 1,2-propylene glycol diacetate, 2 ppm S54V variant, 39 mM sodium percarbonate (85% technical grade, 100 mM effective H202 yield) in 320 mM KP04, pH 7.1 in a total volume of 800 μl. This mixture (40 mM PAA yield) was serially diluted to provide additional mixtures that provide 4.9, 9.9 and 20.5 mM PAA. A mixture with only 400 mM KP04, pH 7.1 was used to determine a total spore count in the absence of PAA. The mixtures were allowed to incubate at room temperature for 3 minutes. A volume of 180 μl of each of the mixtures was then distributed in duplicate cavities of a 96-well round-bottom plate (Costar), containing 20 μl of the spore suspension used in Example 1, to provide a volume total of 200 μl in each cavity. The liquid is gently pipetted 4-5 times to ensure mixing of the components. The mixtures were incubated with the spores for an additional 15 or 30 minutes at room temperature. At the 15 and 30 minute time points, 20 μl of each of the cavities were removed, added to the cavities in a 96-well plate and serially diluted in LB to 107 in a total volume of 100 μl. A volume of 5 μl of each dilution of each spore mixture was spotted on an LA plate, allowed to dry and then incubated overnight at 37 ° C. Also at the 15 and 30 minute time points, and an appropriate volume of each cavity was removed and sufficiently diluted in dH20 to provide a measurable amount of PAA using the ABTS scale test with a standard, as described in Example 1. The results of these tests are shown in Table 2. The results of spore removal are represented as an average of the duplicates.

Elimination of Spores in Detergent. The experiment was repeated exactly as described except that a 1: 500 dilution of Purex in 320 mM KP04, pH 7.1, was used in place of the buffer. The results were presented in Table 3 (average of duplicates). The controls include several reaction components in 400 mM buffer of KP04, pH 7.1: 2 ppm of variant S54V, 2 ppm of • S54V variant with 39 mM percarbonate, 100 mM of 1,2-propylene glycol diacetate, 100 mM of 1,2-propylene glycol diacetate with 39 mM sodium percarbonate, 39 mM sodium percarbonate. All these treatments provide equivalent levels of spores / ml after 30 minutes of incubation, except sodium percarbonate alone (lxlO9 spores / ml against 5xl09 spores / ml for other controls). This decrease was not observed with sodium percarbonate in combination with other components and of course it is not dramatic as the removal observed by mixing all 3 components at comparable levels (100% elimination).

EXAMPLE 4 Enzymatic Generation of PAA to Eliminate Trlchodepna Spores. reesei In this Example, conducted experiments are described to evaluate the capacity of elimination of PAA in spores of T. reesei. Spores of T. reesei were prepared by growing the strain for approximately 4 days in Papa Dextrose (PDA) medium at 30 ° C. When the plaque is approximately 75% covered by growth of fungus, it was incubated at room temperature for seven days until there is confluent growth. Spores scraped off the plate using a cotton tipped swab, resuspended in 1 ml of 10% glycerol and frozen at -80 ° C until use. Before use in the spore removal assay, the spore suspension was thawed, the spores formed into pellets by centrifugation, washed twice with 1 ml of dH20, and resuspended in 1 ml of dH20. The spore removal experiments were carried out as described in Example 3, except that 20 μl of fungal spore preparation was added to the cavities of the 96-well plate instead of the Bacillus spores. Also, the mixtures were made in such a way that the amount of peracid generated was 40, 13.3, 4.4 and 1.5 mM. Dilutions of the 15 and 30 minute incubations were plated on PDA medium. The actual amount of generated PAA was determined as described in Example 1, at the 15 and 30 minute time points. The results are presented in Table 4. These results indicate that the fungal spores were eliminated by PAA generated by the AcT system and at a lower PAA level than the B spores. subtilis.

EXAMPLE 5 Production of Peracid Acid from Glucose and Propylene Glycol Diacetate In this example, experiments are described to assess the amount of peracetic acid produced from glucose and propylene glycol diacetate. A 15 ml solution containing 50 mM KP04, pH 7.1 with 60 mM glucose, and 20 mM 1,2-propylene glycol diacetate (Aldrich) was prepared. The solution is continuously sprayed with air and stirred at room temperature. The reaction to generate H202 is initiated by the addition of 100 units of glucose oxidase (Oxigen HP, Genencor International) and allowed to continue for 1 hour. A sample was removed and tested for PAA prior to the addition of 2 ppm S54V variant, to initiate PAA production from the formed H202. The results are presented in Figure 2. Two additional samples were removed at the times indicated in Figure 2, and the PAA concentration determined as described above. No PAA was detected before the addition of enzymes and approximately 9.5 mM of PAA is produced from 20 mM of 1,2-propylene glycol diacetate and H202 is produced from the glucose / glucose oxidase reaction.

EXAMPLE 6 Generation of Peracetic Acid by AcT at Different Temperatures In this example, conducted experiments are described to assess the generation of paracetic acid by AcT at different temperatures. Such generation provides means to solve problems associated with storage instability of PAA at various temperatures. In these experiments, AcT was used to generate PAA over a temperature range from about 20 ° C to about 60 ° C. In some experiments, temperatures such as 21 ° C, 40 ° C and 60 ° C are used. In these experiments, three reactions were prepared, consisting of 320 mM KP04, pH 7.1, 100 mM 1,2-propylene glycol diacetate (Aldrich), and 100 mM sodium percarbonate. The reactions were equilibrated at 21 ° C, 40 ° C and 60 ° C, and then started by the addition of S54V variant to a final concentration of 2 ppm. The results are presented in Figure 3. The samples were removed at the times indicated in the Figure, and the concentration of PAA determined as described above. These results indicate that the enzyme system is functional up to at least 60 ° C.

EXAMPLE 7 Generation of Concentrated Peracetic Acid by AcT In this Example, experiments are conducted to determine the ability of AcT to generate a concentrated solution of peracetic acid. These experiments are conducted to direct the potential benefit to prepare a concentrated peracetic acid solution which is suitable for dosing or diluting in different solutions for use. In this experiment the reaction contains 50 mM KP04, 2 mM H202 (EM Science), 2 M 1,2-propylene glycol diacetate (Aldrich), and the S54V variant at a final concentration of 160 ppm. The reaction was vortexed occasionally to mix the reactants, they are not miscible at this concentration. The mixing was conducted at room temperature. The results are shown in Figure 4. The samples were diluted at the indicated times and the concentration of peracetic acid was determined as described below. All patents and publications mentioned in the specification are indicative of the levels of those skilled in the art to which the invention pertains. All patents and publications are incorporated herein by reference to the same extent as if each individual publication is specifically and individually indicated to be incorporated by reference. In view of the preferred embodiments of the present invention being described, it will be apparent to one of ordinary skill in the art that various modifications can be made to the embodiments described, and that such modifications are intended to be within the scope of the present invention. Those skilled in the art will readily appreciate that the present invention is well adapted to carry out the objects and obtain the purposes and advantages mentioned., as well as those inherent in this. The compositions and methods described herein are representative of preferred embodiments, are exemplary, and are not intended as limitations on the scope of the invention. It will be readily apparent to one skilled in the art that various substitutions and modifications can be made to the invention described herein without departing from the scope and spirit of the invention. The invention illustratively described herein, suitably may be practiced in the absence of any element or elements, limitation or limitations which are not specifically described in this document. The terms and expressions which have been used, are used as terms of description and not of limitation, and there is no intention that in the use of such terms and expressions except any equivalent of the characteristics shown and described or portions thereof, but it is recognized that various modifications are possible within the scope of the claimed invention. Thus, it will be understood that although the present invention has been specifically described by preferred embodiments and optional features, modifications and variations of the concepts herein described by those skilled in the art may be appealed, and that such modifications and variations are considered to be within the scope of the invention. scope of this invention as defined by the appended claims. The invention has been described broadly and generically in this document. Each of the limited species and sub-generic groupings fall within the generic description are also part of the invention. This includes the generic description of the invention with a negative condition or limitation that removes any subject matter of the goods, with respect to whether or not the material removed is specifically mentioned in this document.

It is noted that in relation to this date, the best method known by the applicant to carry out the aforementioned invention, is that which is clear from the present description of the invention.

Claims (58)

1. CLAIMS Having described the invention as above, the content of the following claims is claimed as property: 1. Enzymatic system for generation of peracid in aqueous solution, characterized in that it is suitable for use in decontamination.
2. 2. Enzyme system according to claim 1, characterized in that the system comprises at least one ester substrate, at least one source of hydrogen peroxide, and at least one acyl transferase.
3. 3. Enzyme system according to claim 1, characterized in that the peracid is selected from peracetic acid, pernonanoic acid, perproprionic, perbutanoic, perpentanoic, perhexanoic acid, percents made of long chain fatty acids, and percents made of chain fatty acids short .
4. 4. Enzymatic system according to claim 2, further comprising at least one chemical hydrogen peroxide generating system, characterized in that the chemical hydrogen peroxide generation system comprises at least one chemical selected from sodium percarbonate, perborate, and hydrogen peroxide urea.
5. 5. Enzymatic system according to claim 2, characterized in that it also comprises at least one system for the generation of enzymatic hydrogen peroxide selected from oxidases and their corresponding substrates.
6. 6. Enzyme system according to claim 2, further comprising at least one enzyme hydrogen peroxide generation system, characterized in that it comprises at least one enzyme selected from glucose oxidase, sorbitol oxidase, hexose oxidase, choline oxidase, alcohol oxidase , glycerol oxidase, cholesterol oxidase, pyranose oxidase, carboxyalcohol oxidase, L-amino acid oxidase, glycine oxidase, pyruvate oxidase, glutamate oxidase, sarcosine oxidase, lysine oxidase, lactate oxidase, vanillil oxidase, glycolate oxidase, galactose oxidase, uricase, oxalate oxidase , xanthine oxidase, and wherein the enzyme hydrogen peroxide generation system further comprises at least one suitable substrate for at least one enzyme.
7. 7. Enzyme system according to claim 1, characterized in that it also comprises at least one additional enzyme.
8. 8. Enzyme system according to claim 7, characterized in that at least one additional enzyme is selected from proteases, cellulases, amylases and enzymes that degrade the microbial cell wall.
9. 9. Enzymatic system according to claim 1, characterized in that at least one ester substrate is an ester alcohol.
10. 10. Enzymatic system according to claim 1, characterized in that it also comprises at least one surfactant.
11. 11. Enzymatic system according to claim 1, characterized in that it also comprises at least one detergent.
12. 12. Enzymatic system according to claim 1, characterized in that it is also in a selected form of liquids, granules, foams and emulsions.
13. 13. Method for decontamination, characterized in that it comprises the steps of: a) providing an article in need of decontamination, and at least one system for generation of peracid in aqueous solution, suitable for use in decontamination; b) expose the item to the system under conditions such that the item is decontaminated. 1 .
14. Method according to claim 13, characterized in that the exposure comprises exposing the article to the system under conditions of alkaline or acidic pH.
15. 15. Method according to claim 13, characterized in that the exposure comprises exposing the article to the system under neutral pH conditions.
16. Method according to claim 13, characterized in that the exposure comprises exposing the article to a high temperature.
17. Method according to claim 13, characterized in that the system is in a selected form of liquids, granules, foams and emulsions.
18. 18. Method of compliance with the claim 13, characterized in that the system comprises at least one ester substrate, at least one source of hydrogen peroxide, and at least one acyl transferase.
19. 19. Method according to claim 13, wherein the peracid is selected from peracetic acid, pernonanoic acid, perproprionico, perbutanoico, perpentanoico, perhexanoic acid, peracids made from long fatty chain acid, and peracetic acid produced short chain.
20. 20. Method of compliance with the claim 18, characterized in that it also comprises at least one chemical hydrogen peroxide generation system, selected from sodium percarbonate, perborate and hydrogen urea peroxide.
21. 21. Method of compliance with claim 7 18, characterized in that the system comprises at least one system for the generation of enzymatic hydrogen peroxide selected from oxidases and their corresponding substrates.
22. 22. Method according to claim 18, wherein the system comprises at least one generation system enzymatic hydrogen peroxide is selected from glucose oxidase, sorbitol oxidase, hexose, choline oxidase, alcohol oxidase, glycerol oxidase, cholesterol oxidase, pyranose oxidase carboxialcohol oxidase L-amino acid oxidase glycine, pyruvate oxidase, glutamate, sarcosine oxidase, lysine oxidase, lactate oxidase, vanillyl oxidase, glycolate oxidase, galactose oxidase, uricase, oxalate oxidase, xanthine oxidase, and wherein the enzyme hydrogen peroxide generation system further comprises at least one suitable substrate for at least one enzyme.
23. 23. Method according to claim 13, characterized in that it also comprises at least one enzyme.
24. 24. Method of compliance with the claim 23, characterized in that the enzyme is selected from proteases, amylases, cellulases and enzymes that degrade the microbial cell wall.
25. 25. Method according to claim 13, characterized in that at least one ester substrate is an ester alcohol.
26. 26. Method according to claim 13, characterized in that it also comprises at least one surfactant.
27. 27. Method of compliance with the claim 26, characterized in that the decontamination comprises decontaminating articles contaminated by at least one toxin and / or at least one pathogen.
28. 28. Method according to claim 27, wherein the toxin is selected from botulinum toxin, anthracis toxin, ricin, scombroid toxin, ciguatoxin, tetrodotoxin, and mycotoxins.
29. 29. Method of compliance with the claim 27, characterized in that the pathogen is selected from bacteria, viruses, fungi, parasites and prions.
30. 30. Method according to claim 29, characterized in that at least one pathogen is selected from Bacillus spp. , B, anthracis, Clostridium spp. , C. botulinum, C. perfringens, Listeria spp. , Staphylococcus spp. , Streptococcus spp. , Salmonella spp. , Shigella ssp. , E. coli, Yersinia spp. , Y. pestis, Francisella spp. , F. tularensis, Camplyobacter ssp. , Vibrio spp. , Brucella spp. , Cryptosporidium spp. , Giardia spp. , Cyclospora spp. , Y Trichinella spp.
31. 31. Method according to claim 13, characterized in that the article in need decontamination is selected from hard surfaces, fabrics, food, food products, clothing, binders, carpets, textiles, medical instruments and veterinary instruments.
32. 32. Method according to claim 13, characterized in that the food is selected from fruits, vegetables, fish, shellfish and meat.
33. 33. Method according to claim 31, characterized in that the hard surfaces are selected from domestic surfaces and industrial surfaces. 3 .
34. Method according to claim 33, characterized in that the domestic surfaces are selected from kitchen countertops, sinks, cupboards, chopping boards, tables, shelves, food preparation storage areas, bathroom fixtures, floors, ceilings, walls and bedroom areas.
35. 35. Method according to claim 33, characterized in that the industrial surfaces are selected from areas of food processing, food processing areas, tables, shelves, floors, ceilings, walls, sinks, snacks, airplanes, automobiles. , trains and boats.
36. 36. Method for decontamination, characterized in that it comprises the steps of: 1a) providing an article in need of decontamination, and at least one system for generating peracid in aqueous solution, suitable for use in decontamination; b) generating the peracid in aqueous solution; and c) exposing the article to the peracid in aqueous solution under conditions such that the article is decontaminated.
37. 37. Method according to claim 36, characterized in that the exposure comprises exposing the article to peracid at alkaline or acidic pH conditions.
38. 38. Method according to claim 36, characterized in that the exposure comprises exposing the article to the peracid under neutral pH conditions.
39. 39. Method of compliance with the claim 36, characterized in that the exposure comprises exposing the article to the peracid at elevated temperature.
40. 40. Method according to claim 36, characterized in that the system is in a selected form of liquids, granules, foams and emulsions.
41. 41. Method according to claim 36, characterized in that the system comprises at least one ester substrate, at least one source of hydrogen peroxide, and at least one acyl transferase.
42. 42. Method according to claim 36, characterized in that the peracid is selected from peracetic acid, pernonanoic acid, perproprionic acid, perbutanoic acid, perpentanoic acid and perhexanoic acid and perished elaborated from long chain fatty acids, and percents made from short chain fatty acids .
43. 43. Method according to claim 41, characterized in that the system comprises at least one chemical hydrogen peroxide system selected from sodium percarbonate, perborate, and hydrogen urea peroxide.
44. 44. Method according to claim 41, characterized in that the system comprises at least one system for the generation of enzymatic hydrogen peroxide selected from oxidases and their corresponding substrates.
45. 45. Method of compliance with the claim 41, characterized in that it also comprises at least one peroxide generation system selected from glucose oxidase, sorbitol oxidase, hexose oxidase, choline oxidase, alcohol oxidase, glycerol oxidase, cholesterol oxidase, pyranose oxidase, carboxyalcohol oxidase, L-amino acid oxidase, glycine oxidase, pyruvate oxidase, glutamate oxidase, sarcosine oxidase, lysine oxidase, lactate oxidase, vanillyl oxidase, glycolate oxidase, galactose oxidase, uricase, oxalate oxidase, xanthine oxidase, and wherein the system for the generation of enzymatic hydrogen peroxide also comprises less, a suitable substrate by at least one enzyme.
46. 46. Method according to claim 36, characterized in that it also comprises at least one enzyme.
47. 47. Method according to claim 46, characterized in that at least one enzyme is selected from proteases, amylases, celluloses, and enzymes that degrade the microbial cell wall.
48. 48. Method of compliance with the claim 36, characterized in that at least one ester substrate is an ester alcohol.
49. 49. Method according to claim 36, characterized in that it also comprises at least one surfactant.
50. 50. Method according to claim 36, characterized in that the decontamination comprises decontaminated articles, contaminated by at least one toxin and / or at least one pathogen.
51. 51. Method of compliance with the claim 50, characterized in that the toxin is selected from botulinum toxin, anthracis toxin, ricin, scombroid toxin, ciguatoxin, tetrodotoxin and mycotoxins.
52. 52. Method according to claim 50, characterized in that the pathogen is selected from bacteria, viruses, fungi, parasites and prions.
53. 53. Method according to claim 52, characterized in that at least one pathogen is selected from Bacillus spp. , B. Anthracis, Clostridium spp. , C. botulinum, C. perfringens, Listeria spp. , Staphylococcus spp. , Streptococcus spp., Salmonella spp. , Shigella ssp. , E. coli, Yersinia spp. , Y. pestis, Francisella spp. , F. tularensis, Camplyobacter ssp. , Vibrio spp. , Brucella spp. , Cryptosporidium spp. , Giardia spp. , Cyclospora spp. , and Trichinella spp.
54. 54. Method according to claim 36, characterized in that the article in need of decontamination is selected from hard surfaces, fabrics, foodstuffs, foodstuffs, garments, rugs, folders, textiles, medical instruments and veterinary instruments.
55. 55. Method according to claim 36, characterized in that the food is selected from fruits, vegetables, fish, seafood and meat.
56. 56. Method of compliance with the claim 55, characterized in that the hard surfaces are selected from domestic surfaces and industrial surfaces.
57. 57. Method of compliance with the claim 56, characterized in that the domestic surfaces are selected from kitchen countertops, sinks, cupboards, chopping boards, tables, shelves, food preparation storage areas, bathroom appliances, floors, ceilings, walls and bedroom areas.
58. 58. Method according to claim 56, characterized in that the industrial surfaces are selected from food processing areas, food processing areas, tables, shelves, floors, ceilings, walls, sinks, snacks, airplanes, automobiles. , trains and boats.