Detergent with stabilised enzyme systems

FIELD: chemistry.

SUBSTANCE: invention relates to a cleaning, bleaching or disinfecting composition which is a stabilised oxidase composition containing said oxidase and a stabiliser and at least one substrate for said oxidase, where said stabiliser contains at least one oxidase inhibitor, where said stabiliser is selected from thiosulphate and 2-amino-2-methyl-1-propanol. In certain especially preferable versions of the invention, said oxidase is selected from glucose oxidase, sorbitol oxidase, choline oxidase, hexose oxidase and alcohol oxidase. The invention also relates to a method of forming a bleaching product in a detergent solution, involving a step for adding said composition to said detergent solution, wherein the preferred version, said bleaching product is a peroxide or a bleaching system which can be activated using peroxide.

EFFECT: ensuring stability a H2O2-generating system during storage and production of bleaching agents when diluting a detergent in a detergent solution.

15 cl, 8 tbl, 9 ex, 3 dwg

 

This application claims priority under pending provisional patent application U.S. serial number 60/818824 filed July 6, 2006

The present invention provides methods and compositions for stabilization of enzymes oxidase during storage. In some preferred embodiments of the invention oxidase is a component of liquid compositions which additionally contain at least one oxidase substrate. In some preferred embodiments of the invention oxidase is the component of the liquid detergent (detergent) compositions. In some especially preferred embodiments of the invention oxidase stabilized by adding to the liquid cleaning medium reversible inhibitor oxidase. In some especially preferred embodiments of the invention oxidase stabilize bisulfite. In additional preferred embodiments of the invention the use of a reversible inhibitor also prevents premature formation of peroxide during storage of liquid detergent. In additional embodiments the invention, the liquid preparations of detergent consisting of an oxidase enzyme, its substrate and its reversible inhibitor, at a dilution liquid is about detergent in the washing solution for washing produce a product with active oxygen (peroxide).

Detergents for Laundry and dishes consist of complex mixtures containing a wide range of ingredients, which usually include a number of components, such as ionic and nonionic surfactants, solvents, builders, perfumes, enzymes and bleaching components. For such complex mixtures is well known problems associated with the storage stability, in particular stability of the enzymes. In some cases, the stability problems associated with the physical stability of detergent, while in other cases they are associated with the functional stability of the individual ingredients of the detergent. Enzymes, such as oxidases, in particular, is subject to restrictions stability when stored in the liquid detergent. This prevents their widespread use in the cleaning compositions intended for fabrics and for home use, which refers to the bleaching action. The preservation of the enzymatic activity of the oxidase in detergents during storage is a difficult task, especially in detergents, which also contain components in the form of the oxidase substrate. The presence of oxidase, oxidase and substrate leads to the formation of hydrogen peroxide directly in situ. Peroxide, bodoro is and reduces the stability of the enzyme by oxidation enzymes in liquid and dry preparations. It is believed that the destruction of the enzymes under the action of the peroxide is carried out through various mechanisms (e.g., oxidation of essential amino acid residues in the enzyme through interactions with cofactors of enzymes and so on). However, it is believed that the present invention is not limited to any particular mechanism. However, the destruction of enzymes by the action of peroxide often leads to a gradual loss of activity. In dry preparations of detergent enzymes can be stabilized (for example, by using encapsulation of enzymes, as described in the application WO 96/02623 included in this description in its entirety by reference).

In this area there are various bleach for washing and activators washing (see, for example, the publication of Grime and Clauss, Chem. Indust., 20:647-649, 652-653 [1990]; Sheane and Wilkinson, Tinctoria 101:36-41 [2004]; and Broze,Handbook of Detergents,Warwick International, [1999]). Used most often bleaches include perborate sodium, percarbonate sodium, sodium persulfate, perphosphate sodium, urea peroxide, persilat sodium, ammonium, potassium and lithium analogues; in detergents, toothpastes and other products are widely used peroxide, calcium peroxide, zinc peroxide, sodium, carbamoylphenoxy and others, such as sodium hypochlorite and chlorine oxide. Okisliteljno the ability of such peroxides at low temperature can be improved by adding "activator bleaching". In this area there are many bleaching activators, which include acyl compounds such as tetraacetylethylenediamine (TAED), ester compounds such as nonanoyloxybenzenesulfonate (NOBS) and isanonymousallowed (ISONOBS), complexes of transition metals and other compounds.

In this whitening system by adding water during the wash cycle, it generates percolate (e.g., peracetic acid, hydrogen peroxide and/or other related products. Percolate and other products with active oxygen present in the system, then act to whiten or lighten some spots on the tissue or vessel. However, bleaching activators cannot be added to liquid detergents with percarbonates, because they will interact and form percolate and/or other activated oxidizing means. There is therefore a need for a system that produces H2O2that inactive during storage, however, produces hydrogen peroxide during the wash cycle.

Bleach is usually not included in liquid detergents due to poor stability during storage bleaching products in detergents that contain significant amounts of water (for example, more than 1% water). The presence of bleaching products also ESMA adversely affects the storage stability of the enzymes, sensitive to oxidation, and other compounds included in detergents. There is therefore a need for liquid detergents which ensure the generation of bleaching products in situ at a dilution of detergent in the washing solution.

It has been described several oxidase (see, for example, published: Beck et al., Bleach activators. Carbohydrates as Organic Raw Materials III : developed from a Workshop, Wageningen, Nov. 28-29, 1994, pages 295-306 [1996]; Nakayama and Amachi, J. Mol. Catalysis B: Enzymatic 6: 185-198 [1999]; WO 06/008497; WO 05/124012; U.S. patent No. 6399329; WO 01/007555 and WO 03/36094). However, an important limitation of such systems is that when oxidase together with their substrates stored in the liquid detergent, they produce hydrogen peroxide, which in itself can destroy the enzymes, and may also interact with the bleaching activators present in the system. Therefore, such systems oxidase substrate is unstable. In fact, this area remains a need for a tool that provides oxidase, reversible inhibiting in the presence of substrates during storage, which will produce bleach in situ when breeding in the washing solution. In addition, there is the need to obtain bleaching products (for example, products with active oxygen, peroxide and perkiset) at a dilution of detergent in the washing solution for washing the purpose bleach the deposits and/or lighten stains.

As in the case of liquids, the presence of bleaching products in powder detergents often has a strongly negative impact on the stability of the enzymes present in the detergent. Therefore, assumes greater caution in the separation of molecules of enzymes and bleaching products in powdered detergent. Usually this is achieved by separate compilation mixture of enzymes and bleaching products. For example, in some cases, the enzymes get in the form of granulates prepared in such a way as to minimize the penetration of the product with the active oxygen granules containing the enzyme during storage. For such powdered detergent (detergent) systems can also be useful for the application of reversible inhibiting system oxidase enzyme with a substrate.

The present invention provides methods and compositions for stabilization of enzymes oxidase during storage. In some preferred embodiments of the invention oxidase is the component of the liquid detergent compositions. In some especially preferred embodiments of the invention oxidase stabilized by adding to the liquid cleaning medium reversible inhibitor oxidase. In some particularly preferred embodiments, the implementation of izobreteniya stabilize bisulfite. In additional preferred embodiments of the invention the use of a reversible inhibitor also prevents premature formation of peroxide during storage of liquid detergent. In additional embodiments the invention, the liquid preparations of detergent containing oxidase enzyme, its substrate and its reversible inhibitor at a dilution of liquid detergent in the washing solution for washing produce a product with active oxygen (peroxide). In additional embodiments, the implementation of the present invention offers powdered drugs detergent containing at least one enzyme oxidase, at least one oxidase substrate and at least one reversible inhibitor. In some especially preferred embodiments of the invention such powdered preparations of detergent to produce products with active oxygen (peroxide) at a dilution of powdered detergent in the washing solution for washing.

The present invention offers a stable composition oxidase containing at least one oxidase and at least one stabilizer. In some embodiments of the invention, the oxidase is selected from glucose oxidase, corbiculidae, cholinesterase, exotic is easy and alcohol oxidase. In some alternative embodiments the invention, the composition additionally contains at least one substrate at least one oxidase. In some preferred versions of the invention, the substrate is chosen from glucose, lactate, sorbitol, choline, glycerol, ethylene glycol, propylene glycol and ethanol. In some alternative embodiments of the invention at least one stabilizer comprises at least one oxidase inhibitor. In some preferred versions of the invention, the stabilizer comprises at least one sulfite. In some especially preferred embodiments of the invention at least one sulfite selected from hydrosulfite sodium, metabisulfite, sodium and/or sodium bisulfite. In some alternative preferred embodiments of the invention the stabilizer is selected from thiosulfate and 2-amino-2-methyl-1-propanol. In some especially preferred embodiments of the invention the composition is a cleansing, whitening and/or disinfectant composition. In some alternative preferred embodiments of the invention the detergent is a detergent for washing or detergent for dishes. Some of the more embodiments of the invention the detergent is selected from a powder, liquid and gel detergents. In some yet some additional embodiments of the invention the composition is a detergent (detergent) Supplement or product pre-treatment. In yet some additional embodiments of the invention the composition further comprises an activator bleaching or bleach precursor. In some embodiments of the invention, the bleaching activator selected from precursors of percolat, metal complexes, peroxidases and systems acyltransferase substrate. In some particularly preferred embodiments the invention, the composition additionally contains at least one enzyme selected from proteases, amylases, pectinase, pectacles, lipases, mannanase, cellulases, esterases, koutinas, oxidoreductases, hemicellulase and karbohidrat. In some additional embodiments the invention, the composition additionally contains at least one auxiliary ingredient chosen from surfactants, builders, bleaching products, antimicrobial agents, polymers, solvents, salts, buffer funds, chelat forming means, means inhibiting the transfer of dyes, additives for deposition, dispersing agents, enzymes, stabilizers EN zymes is s, catalytic materials, bleach activators, accelerators whitening, previously obtained perkiset, polymeric dispersing means, graziadei tools/anti re-deposition, optical brighteners, soap suds suppressors, dyes, perfumes, tools, giving elasticity to the structure, fabric softeners, carriers, hydrotropes, technological additives, pigments and mixtures thereof.

The present invention also provides methods of obtaining a whitening product in the washing solution, including the state added to the cleaning solution of at least one composition according to the present invention. In yet some additional embodiments of the invention whitening product is a peroxide or whitening system that can be activated using peroxide.

Figure 1 presents a graph showing the effect of bisulfite on the stability and bleaching characteristics of glucose oxidase.

Figure 2 presents a graph showing the effect of bisulfite on the stability and bleaching characteristics of glucose oxidase in the test drives, contaminated blueberries.

Figure 3 presents a graph showing the effect of bisulfite on the stability and bleaching characteristics of the oxidase test tissue samples from numerous what akrasanee in teratomata.

The present invention provides methods and compositions for stabilization of enzymes oxidase during storage. In some preferred embodiments of the invention oxidase is the component of the liquid detergent compositions. In some especially preferred embodiments of the invention oxidase is stabilized by adding to the liquid cleaning medium reversible inhibitor oxidase. In additional preferred embodiments of the invention the use of a reversible inhibitor also prevents premature formation of peroxide during storage of liquid detergent. In additional embodiments the invention, the liquid preparations of detergent containing oxidase enzyme, its substrate and its reversible inhibitor at a dilution of liquid detergent in the washing solution for washing produce a product with active oxygen (peroxide). In some especially preferred embodiments of the invention oxidase stabilized by bisulphite. In additional embodiments, the implementation of the present invention offers liquid preparations of detergent containing at least one oxidase, at least one oxidase substrate and at least one reversible inhibitor. In a particularly preferred variant is ntah the invention, such liquid preparations of detergent at a dilution liquid detergent in the washing solution for washing produce a product with active oxygen (for example, peroxide).

Unless otherwise stated, the implementation of the present invention in practice involves the use of traditional techniques, widely used in molecular biology, Microbiology, protein purification, protein engineering, the determination of the sequences of proteins and DNA, areas of recombinant DNA and industrial application and development of enzymes, all of which are in the competence of the specialist in this field. All patents, patent applications, articles and publications cited in this specification, both above and below, thereby explicitly included in this description by reference.

In addition, provided in the description column are not limitations of the various aspects or embodiments of the invention, which can be obtained by reference to the description of the invention in General. Accordingly, the terms defined directly below are more fully defined by reference to the description of the invention in its entirety. However, to facilitate understanding of the invention, the following are the definitions of some terms.

Unless otherwise noted, all materials used in this description of the technical and scientific terms have the same meaning, which is widely understood to a person skilled in the art to which this invention. Although any methods and materials, the such or equivalent described in this description, will be used when implementing the present invention in practice, in the present description describes the preferred methods and materials. Accordingly, the terms defined directly below are more fully described by reference to the description of the invention in its entirety. Also used the singular number include reference to the plural, unless the context clearly indicates a different. Unless otherwise indicated, nucleic acids are written left-to-right orientation from 5' to 3'; amino acid sequences are written left to right in the direction from the amino group to carboxypropyl, respectively. It should be understood that the invention is not limited to the particular methodology, protocols, and described reagents, which may vary depending on the context in which they are used by experts in this field.

It is believed that every maximum numerical limitation given throughout this description of the invention, includes any numerical limitation of a lower order, as if such numerical limits lower order were described in this description, explicitly. Each of the minimum numerical limitation given throughout this specification the invention will include any of isleno restriction of a higher order, as if such numerical limitations of a higher order have been described in this description, explicitly. Each numerical range given throughout this description of the invention will include any narrower numerical range that falls within such broader numerical range, as if such narrower numerical ranges have been described in this description, explicitly.

Used in this description, the term "oxidase" refers to enzymes that catalyze the oxidation reaction/recovery, including molecular oxygen (O2) as the electron acceptor. In such reactions, the oxygen is reduced to water (H2O) or hydrogen peroxide (H2O2). Oxidases are a subclass of oxidoreductases.

Used in this description, the term "glucose oxidase" ("Gox") refers to the enzyme oxidase (EC 1.1.3.4), which catalyzes the oxidation of β-D-glucose to D-glucono-l,5-lactone, which is then hydrolyzed to gluconic acid with concomitant reduction of molecular oxygen to hydrogen peroxide.

Used in this description, the term "alcohol oxidase" ("Aox") refers to the enzyme oxidase (EC 1.1.3.13), which converts the alcohol to the aldehyde with a concomitant reduction of molecular acid is kind to hydrogen peroxide.

Used in this description, the term "cholinesterase" ("Cox") refers to the enzyme oxidase (EC 1.1.3.17), which catalyzes the oxidation of choline to glycinebetaine with the transfer of four electrons and betanova aldehyde as an intermediate product and a concomitant recovery of two molecules of molecular oxygen to two molecules of hydrogen peroxide.

Used in this description, the term "exotoxins" ("Hox") refers to the enzyme oxidase (EC 1.1.3.5) for oxidation of mono - and disaccharides to their corresponding lactones with a concomitant reduction of molecular oxygen to hydrogen peroxide. Hexosaminidase able to oxidize a variety of substrates, including D-glucose, D-galactose, maltose, cellobiose and lactose, etc. it Is believed that the present invention is not limited to any specific hexoses.

Used in this description, the term "glycerinated" refers to the enzyme oxidase (EC 1.1.3.), which catalyzes the oxidation of glycerol to glyceraldehyde with concomitant reduction of molecular oxygen to hydrogen peroxide.

Used in this description, the term "corbiculidae" refers to the enzyme oxidase polyols (EC 1.1.3.), which catalyzes the oxidation of the substrate (for example, D-sorbitol) to D-glucose with concomitant reduction of molecular oxygen to the PE the oxide of hydrogen. The substrates for corbiculidae also include various polyols (for example, xylitol, Arabic, mannitol, ribitol, glycerin, propandiol and propylene glycol). Used in this description, the term "polyol" refers to chemical compounds that contain many hydroxyl groups.

In the present invention find application additional oxidase, including, but not limited to, cholesterol oxidase, pianosounds, carboxypropyl oxidase, L-amino acid oxidase, glycerokinase, piruwatkinaza, glutaredoxins, sarcosinates, lazinesses, lacticacid, vanillylamide, glycosidase, galactosidase, uricase, oxalidaceae, xanthine oxidase.

Used in this description, the term "inhibitor" refers to chemical compounds that can reduce or block the catalytic activity of the enzyme. In particularly preferred versions of the invention, the inhibitors reduce or block the catalytic activity of at least one oxidase. Examples of the oxidase inhibitors include acetate, salts of silver, halide ions, secondary and tertiary alcohols, isocyanates, isothiocyanate, analogues of glucose, bisulfite, sulfite, thiosulfate, metabisulfite, zinc salts, diethylcarbamyl, methylmethanesulfonate, Acrylonitrile, 2-amino-2-methyl-1-propanol.

COI is Leshey in this description, the term "reversible inhibitor of the enzyme" refers to molecules, they join to the enzyme and reduce the reaction speed with his participation. In some embodiments of the invention are reversible inhibitors of enzymes are affected by changes in the concentration of enzyme substrate depending on the inhibitor. In some embodiments of the invention are reversible inhibitors of enzymes attached to the enzyme through weak ties that such ties used to attach to the substrate. Thus, a reversible inhibitor blocks the enzyme is not forever, by removal of the inhibitor from the enzyme has the opportunity to join its substrate and to share their substrate. In some embodiments of the invention a reversible enzyme inhibitors are competitive inhibitors that ecovalence interact with the enzyme and/or compete with the substrate for the active site of the enzyme, and/or have a structure similar to the substrate, products and/or transitional state. In additional embodiments the invention, the reversible inhibitor is a non-competing inhibitor of the enzyme that joins in on the plot, the presence of the enzyme and different from the active site, and/or causes conformational changes of the enzyme, which reduce and/or block catalyticallyactive. It is believed that this term is not limited to any particular mechanism or type reversible inhibitor of the enzyme. A necessary condition is only that the effects of an inhibitor of the enzyme were reversibly to the enzyme functioned in the absence of inhibitor and/or in the absence of the effects of the inhibitor.

Used in this description, the term "compatible" means that the materials cleaning compositions do not reduce the enzymatic activity of the proposed enzyme(s) oxidase to such an extent that a oxidase(s) becomes inefficient, as it is desirable in situations in General use. Materials specific cleansing composition will be detailed in the example below.

Used in this description, the term "effective amount of the enzyme" refers to the amount of enzyme necessary to achieve the enzymatic activity provided for in a particular application. Such effective amounts can be easily determined by a person skilled in the art and is based on many factors, such as a specific variant of enzyme used, the use of purification, the specific composition of the cleaning composition, requires a liquid or dry (e.g., granulated) composition, etc.

Used in this description, the phrase "stability of detergent" refers to the stability to the position of detergent. In some embodiments of the invention, the stability is measured during the application of detergent, while in other embodiments of the invention the term refers to the stability of the detergent composition during storage.

The term "improved stability" is used to show the best stability of the enzymes in the compositions containing the substrate. In preferred embodiments of the invention in detergents inhibitors intended for washing or caring for utensils, enzymes have improved stability during storage compared with the corresponding drugs without enzyme inhibitors. In preferred embodiments of the invention, the system enzyme/substrate in detergents inhibitors intended for washing or caring for utensils and has improved stability during storage compared with the corresponding drugs without enzyme inhibitors.

Used in this description, the term "stability under oxidation" refers to the ability of proteins to function in oxidative conditions. In particular, the term refers to the ability of proteins to function in the presence of various concentrations of H2O2, perkiset and other oxidants. Stability at different on ikitelli conditions can be measured either by using standard techniques, well-known experts in this field, and/or using the steps described in this way. A significant change in the stability of the oxidation is confirmed by the increase or decrease (in most embodiments of the invention, it is preferable to increase) at least about 5% or more of the half-life of enzymatic activity compared to the enzyme activity observed in the absence of oxidizing compounds.

Used in this description, the term "stability when changing pH" refers to the ability of proteins to function at a specific pH. As a rule, most enzymes have a limited pH range in which they will operate. In addition to the enzymes that function in the middle range of pH (about pH 7), there are enzymes that are able to work in conditions with very high or very low pH. Stability at different pH can be measured either using standard techniques known to experts in the field, and/or using the steps described in this way. A significant change in stability at a pH change is confirmed by the increase or decrease (in most embodiments of the invention, it is preferable to increase) at least about 5% and is more than half the enzymatic activity compared to the enzyme activity at the optimum pH for enzymes. However, it is believed that the present invention is not limited by any level of stability when changing pH or pH range.

Used in this description, the term "thermal stability" refers to the ability of proteins to function at a specific temperature. As a rule, most of the enzymes has a limited range of temperatures at which they will operate. In addition to the enzymes that work in the moderate temperature range (for example, at room temperature), there are enzymes that can operate at very high or very low temperatures. Thermal stability can be measured either by using known techniques, or using the steps described in this way. A significant change in thermal stability is confirmed by the increase or decrease (in most embodiments of the invention, it is preferable to increase) at least about 5%, or more than half of catalytic activity of the mutant when it is exposed to a predetermined temperature. However, it is believed that the present invention is not limited to any level of stability at any temperature or temperature range.

Used in this description, the term "chemical stability" refers to stabilnosti (for example, enzyme) with respect to chemical substances that can have a negative impact on their activity. In some embodiments of the invention such chemicals include, but are not limited to, hydrogen peroxide, percolate, anionic detergents, cationogenic detergents, nonionic detergents, chelating tools, etc. But it is believed that the present invention is not limited to any particular level of chemical stability or range of chemical stability.

Used in this description, the terms "purified" and "isolated" refer to the removal of contaminants from a sample. For example, the enzyme of interest, purified by removal of contaminating proteins and other compounds that are not enzyme of interest from a solution or upon receipt. In some embodiments of the invention the recombinant enzymes of interest is expressed in a bacterial or fungal host cells, and these recombinant enzymes of interest are purified by removing other components of the host cell; the percent of recombinant enzyme of interest, among the polypeptides in the sample increases.

Used in this description, the term "proteins, representing the nteres" refers to proteins (for example, the enzyme or enzyme of interest")that are subject to analysis, identification and/or modification. In the present invention are the use of natural and recombinant (e.g., mutant) proteins.

Used in this description, the term "protein" refers to any composition containing amino acids and identified by a person skilled in the art as protein. The terms "protein", "peptide" and " polypeptide used in this specification interchangeably. When the peptide is part of a protein, the person skilled in the art the use of this term will become clear in the context.

As used in this description, the term "cleaning composition" and "cleaning agents" refer to compositions that find use in the removal of undesirable compounds from the products subjected to purification such as fabric, dishes, contact lenses, other solid substrates, removing unwanted compounds from the hair (shampoos), skin (Soaps and creams), teeth (liquid mouth rinse, toothpaste, and so on, the Term includes any materials/compounds selected for the particular type of desired cleansing composition, and product form (e.g., liquid, gel-like composition, the composition is in the form of granules or spray), provided that the composition is compatible with the oxidase and the other enzyme(s), the use of which has been created in the composition, and any reversible inhibitors of enzymes in the composition. The specific choice of materials cleansing compositions are easy to make with regard to the surface of the product or fabric being cleaned, and the desired form of the composition for use in cleaning.

In addition, the terms refer to any composition that is suitable for cleaning, bleaching, disinfection and/or sterilization of any object and/or surface. It is believed that the terms include, but are not limited to, detergent compositions (e.g., liquid and/or solid detergents for washing and detergents for fine fabrics; cleaning preparations for hard surfaces, such as glass, wood, ceramic and metal table tops and Windows; cleaning products for carpets; cleaning products for ovens; air fresheners fabrics; fabrics softeners and removers for pre-stain removal Laundry and textiles, and detergents for dishes).

Moreover, as used in this description, the term "cleaning composition" includes, unless otherwise specified, detergent General purpose in granulated or powdered form or detergent designed for intensive cleaning mode, in particular cleansing detergent; detergent General purpose liquid, gel-like sludge is pasty form, in particular, the so-called types of fluids, designed for intensive cleaning mode (HDL); liquid detergents for fine fabrics; means for manual washing of dishes or tools for easy washing, in particular, the means related to the type of rich foaming; means for machine washing, including different types in the form of tablets, granules, liquids and additives for rinsing, designed for home use and use in institutions; liquid cleaning and disinfecting agents, including antibacterial types cleanser, pieces of cleansing soap, liquid for rinsing the oral cavity cleaning tool for dentures, shampoos for vehicles or carpets, cleaning products for the bathroom; shampoos and conditioners for hair; shower gels and foam baths, cleaners for metals; as well as cleansing tools such as bleaching additives and funds type of pencil to remove stains or pre-processing.

The term "detergent composition" and "the preparation of detergent are applicable for mixtures that are intended for use in cleaning the environment for cleanup of contaminated sites. In some preferred embodiments of the invention, the term applies to washing ton is Anya and/or clothing (e.g., "detergents for washing"). In alternative embodiments of the invention the term refers to other detergents, such as detergents used for cleaning utensils, Cutlery, etc. (for example, detergents for washing dishes"). It is believed that the present invention is not limited to any particular drug detergent or composition. Furthermore, it is believed that in addition to perhydrol the term includes detergents that contain surfactants, transferase(s), hydrolytic enzymes, oxidoreductase, builders, bleaching agents, bleaching activators, podsinwowa tools and fluorescent dyes, anti-caking agents, masking tools, enzyme activators, enzyme inhibitors, antioxidants and soljubilizatory. In some preferred versions of the invention, the preparations of detergent include, but are not limited to, tools, published in patent applications U.S. serial No. 10/576331 and 10/581014 and applications WO 05/52161 and WO 05/056782, found use in the present invention. However, it is believed that the present invention is not limited to any particular drug detergent(mi), as in the present invention finds application in any suitable drug my what it means.

Used in this description, the term "composition for washing " refers to all forms of compositions for cleaning the utensils, including flatware, including, but not limited to, granular and liquid forms. It is believed that the present invention is not limited to any particular type or composition for dishes. Moreover, the present invention finds use when cleaning utensils (e.g., tableware, including, but not limited to, plates, cups, glasses, bowls etc) and Cutlery (for example, devices including, but not limited to, spoons, knives, forks, utensils for serving and so on) of any material, including, but not limited to, ceramics, plastics, metals, porcelain, glass, acrylic materials, etc. the Term "ware" applies as crockery and Cutlery.

Used in this description, the term "detergent characteristics" of an enzyme refers to the contribution of the enzyme in the wash, which provides cleaning means extra cleaning performance compared to detergent without the addition of enzyme to the composition. Cleaning characteristics are compared in the respective conditions of washing.

The term "appropriate wash conditions" used in this description in order to specify conditions, in particular temperature, washing time, washing the e mechanisms the concentration of the soap solution, the type of detergent and the hardness of the water is actually applied in the market segment detergent designed for home use.

The term "improved cleaning characteristics" is used to indicate that when removing stains from products (e.g., tissue or utensils and/or Cutlery), washed in appropriate circumstances wash, get the best end result or fewer of the enzyme, based on mass, to obtain the same end result as compared with another enzyme.

The term "not changing detergents characteristics" is used to indicate that the detergent characteristics of the enzyme, based on the weight, amount to at least 80% relative to another enzyme in appropriate circumstances, wash.

Detergent characteristics of enzymes is usually measured by their ability to remove certain specific spots in the respective test conditions. In these test systems can be adjusted to other relevant factors such as the composition of the detergent composition, the concentration of the soap solution, water hardness, detergent machinery, time, pH and/or temperature, thus, to simulate the conditions typical for home use in a specific market segment.

Used in Yes the Mr. the description, the term "disinfectant" refers to the removal of dirt from surfaces and also to the inhibition or destruction of microbes on the surfaces of products. It is believed that the present invention is not limited to any particular surface, product, or contamination(s), or microbes to be deleted.

Cleaning and detergent preparations

Detergent compositions of the present invention are offered in any suitable form, including, for example, but not limited to, liquids, granules, emulsions, gels and pastes. When using a solid detergent composition, the detergent is preferably manufactured in the form of granules. The granules preferably receive additional content protective equipment (see, for example, patent application U.S. serial No. 07/642669, filed January 17, 1991, included in this description by reference). Similarly, in some embodiments of the invention the granules are produced such that they contain materials designed to reduce the dissolution rate of the pellets in the scrubbing medium (see, for example, U.S. patent No. 5254283 included in this description in its entirety by reference). In addition, the enzymes of the present invention find use in drugs, in which the substrate and the enzyme are in the same granule. Thus, in some embodiments of the invention the efficiency of the enzyme, the Pris is concerned in the preparation, is increased by providing a high local concentration of enzyme and substrate (see, for example, the publication of the patent application U.S. US2003/0191033 included in this description by reference). In the present invention finds application in any suitable drug and/or drugs (see, for example, U.S. patent No. 5204015 included in this description by reference). Specialists in this area is well known for other drugs that are used as cleaning compositions.

In addition, proteins, in particular stabilized oxidase of the present invention, can be incorporated into the formulation known powdered and liquid detergents having pH from 3 to 12.0, at a content of about 0.001 to about 5% (preferably from 0.1 to 0.5%) by weight.

It is assumed that stable oxidase of the present invention will find application in any suitable cleansing compositions, including, but not limited to, use in Kuskovo and liquid Soaps, preparations for the care of the dishes, use for surface cleaning, cleaning solutions or products for contact lenses, processing waste, application for textiles, for bleaching pulp, disinfecting substances for skin care, oral care mouth, hair care, etc.

Despite the fact that this is not what is required from the point of view of the purposes of the present invention, in further illustrates a non-limiting list of supporting materials suitable for use in the instant cleaning compositions and which are used in certain embodiments of the invention, for example, to support or enhance cleaning performance, for treatment of the substrate to be cleaned, or to modify the aesthetic properties of the cleansing composition, as in the case of perfumes, pigments, dyes or the like, it is Clear that such auxiliary materials are used in addition to stable enzymes of the present invention. The precise nature of these additional components, and levels of their inclusion depends on the physical form of the composition and nature of the cleaning operation for which it should be applied. Suitable support materials include, but are not limited to, surfactants, builders, hepatoblastoma tools, tools, inhibiting migration of the dye precipitating additives, dispersing funds, additional enzymes, and enzyme stabilizers, catalytic materials, bleach activators, accelerators, bleach, prior percolate, polymeric dispersing means, gryazeudalyayuschaya tools/anti-re-deposition, optical brighteners, suppressors m is through the foam, dyes, fragrances, tools, giving elasticity to the structure, the fabrics softeners, carriers, hydrotropes, processing AIDS and/or pigments (see, for example, U.S. patent No. 5576282, 6306812 and 6326348 included in this description by reference). The above auxiliary ingredients can make the balance cleansing compositions of the present invention.

In some preferred embodiments of the invention in detergent compositions of the present invention uses a surface-active agent (i.e. surfactant, including anionic, nonionic and ampholytic surfactants for use in detergent compositions is well known. Some surfactants suitable for use in the present invention, is described in British patent application No. 2094826 A, included in this description by reference. In some embodiments, implementation of the present invention apply a mixture of surfactants. For example, the number of known compounds are suitable surfactants applicable to compositions containing mutant proteins according to the invention. These include nonionic, anionic, cationogenic, anionic or zwitterionic detergents (see, for example, U.S. patent No. 404128 and 4261868).

Anionic surfactants suitable for use in detergent compositions of the present invention, include alkylbenzenesulfonate with a linear or branched chain; sulfates simple alilovic or alkenilovyh esters containing alkyl groups or alkeneamine group with a linear or branched chain; alkyl sulphates or alkanesulfonyl; reincorporate; alkanesulfonyl etc. Suitable counterions for anionic surfactants include alkali metal ions such as sodium and potassium; alkaline earth metal ions such as calcium and magnesium; ammonium ion; alkanolamine containing from 1 to 3 alkanols groups with the number of carbon atoms is 2 or 3.

Ampholytic surfactants, which are used in the present invention include sulfate Quaternary ammonium salts, ampholytic surfactants of betaine type, etc. of Such ampholytic surfactants have both positively and negatively charged groups in the same molecule.

Nonionic surfactants, which are used in the present invention usually contain simple polyoxyalkylene esters, and alkanolamine higher fatty acids or their adducts with alkalization, glycerin complex is monetary fatty acids, etc.

In some preferred versions of the invention, the surfactant or mixture of surfactants included in the detergent compositions of the present invention, used in amounts of from about 1 wt.% to about 95 wt.% of the total weight of the detergent composition and preferably from about 5 wt.% to about 45 wt.% by weight of the total detergent composition. In various embodiments of the invention in the compositions of the present invention includes numerous other components. It is believed that the present invention is not limited to the specific above examples. Moreover, it is believed that this invention will find application additional connections.

In some embodiments of the invention offer cleansing compositions contain at least one chelate forming agent. Fit hepatoblastoma tools include, but are not limited to, hepatoblastoma tools for copper, iron and/or manganese, and mixtures thereof. If applied chelate forming agent, a cleansing composition typically contains from about 0.1 to about 15% or even from about 3.0 to about 10% chelat forming means based on the weight of the tested cleansing composition.

In some embodiments of the invention the cleaning composition of the present invention contain an additive for deposition. Suitable additives for deposition include, but are not limited to, polyethylene glycol, polypropyleneglycol, polycarboxylate, soil release polymers, such as polyterephthalic acid, clays, such as kaolinite, montmorillonite, atapulgite, illite, bentonite, halloysite and mixtures thereof.

In some additional embodiments of the invention the cleaning composition of the present invention can also include one or more means of inhibiting the migration of the dye. Suitable polymeric preparations inhibiting the transfer of dyes include, but are not limited to, polyvinylpyrrolidone polymers, polyamine N-oxide polymers, copolymers of N-vinylpyrrolidone and N-vinylimidazole, polivinilatsetatny and polyvinylimidazole or mixtures thereof. If test cleansing compositions have means, inhibiting migration of the dye, they are usually present in amounts of from about 0.0001 to about 10%, from about 0.01 to about 5% or even from about 0.1 to about 3% by weight based on the weight of the cleansing composition.

In some additional embodiments of the invention the cleaning composition of us who oedema invention also contain dispersing means. Suitable water-soluble organic materials include Homo - or copolymer acids or their salts in which the polycarboxylic acid comprises at least two carboxyl radicals separated from each other by no more than two carbon atoms.

In some embodiments of the invention these detergent cleaning compositions optionally include other enzymes, which usually provide cleaning performance and/or warranty of fabric care. Examples of suitable enzymes include, but are not limited to, hemicellulase, peroxidase, protease, cellulase, xylanase, lipase, phospholipase, esterase, cutinase, pectinase, pectolyase, keratinase, reductase, oxidase, oxidoreductase, peroxidase, lipoxygenase, ligninase, mannanase, pullulanase, tannaz, pentosanase, peroxidase, Malagasy, β-glucanase, arabinosidases, hyaluronidase, chondroitinase, laccase, endoglycosidase, and amylases, or mixtures thereof. A typical combination is a blend of traditionally used type enzymes protease, lipase, cutinase and/or cellulase in conjunction with amylase.

Adding protein to conventional cleaning compositions does not create any special limitations. In other words, any temperature and pH suitable for the detergent is also suitable for the present is omposite provided what pH is in the range in which the enzyme(s) is active and the temperature below denaturirovannyj described proteins. In addition, proteins of the invention find use in cleaning, bleaching and disinfecting compositions without detergents, this time either in pure form or in combination with a source of hydrogen peroxide, ester substrate (for example, either added to, or characteristic of the applied system, such as system patches that contain esters, the pulp, which contains esters, etc), other enzymes, surfactants, builders, stabilizers, etc. moreover, it is believed that the present invention is not limited to any specific product or application.

In some additional embodiments of the invention the cleaning composition of the present invention include catalytic metal complexes. One of the types of metal-containing bleaching catalyst is a catalyst system containing a transition metal cation, determining the catalytic activity of bleach, such as the cations of copper, iron, titanium, ruthenium, tungsten, molybdenum, or manganese; additional metal cation with a lower catalytic activity bleach or without the cat the lytic activity of the bleach, such as the cations of zinc or aluminum, and complexing agents with defined stability constants for the catalytic cations of metals and additional metals, particularly ethylenediaminetetraacetic acid, ethylenediaminetetra(methylenephosphonate acid) and their soluble salts. Such catalysts are described in U.S. patent No. 4430243 included in this description by reference. In some embodiments of the invention for the catalysis used in the compositions of the connection is used manganese. Such compounds and levels of use are well known in the art (see, for example, U.S. patent No. 5576282). In some alternative embodiments for carrying out the invention known applicable in this invention cobalt catalysts bleaching (see, for example, U.S. patent No. 5597936, 5595967, 5597936 and 5595967).

In some embodiments the invention, the cleaning compositions optionally contain a complex of transition metals with macropolicies rigid ligand ("MRL"). In practice, and without any limitation of the compositions and cleaning processes of the present invention can be adjusted to provide a concentration of active product MRL in the aqueous washing medium about at least one part in a hundred million, and preferably will be provided with the contents of the MRL in the washing solution is approximately 0.005 h/m to approximately 25 hours/million more preferably approximately from 0.05 h/m to approximately 10 hours per million, and most preferably approximately from 0.1 h/m to approximately 5 hours/million Preferred transition metals in a soluble bleaching catalyst based on transition metals include manganese, iron and chromium. Preferred MRL are special types of rigidity of the ligand, which is a cross-bridge, such as 5,12-diethyl-1,5,8,12-tetraazabicyclo[6.6.2]hexadecane. Suitable MRL for transition metals easily get through known in the field of methods (see, for example, application WO 00/332601 and U.S. patent No. 6225464).

In some embodiments of the invention the cleaning composition of the present invention contain one or more of detergent builders or system builders. When applied the amendment, which is to be tested cleansing composition typically contains at least about 1%, from about 3% to about 60%, or even from about 5% to about 40% amendment in the calculation of the mass of the tested cleansing composition. The builders include, but are not limited to, salts of polyphosphates, alkali metal, ammonium and alkanolamine, silicates of alkali metals, carbonates of alkaline earth and alkali metal, p is dicarboxylate connection of aluminosilicate builders, hydroxypolycarboxylic ethers, copolymers of maleic anhydride with ethylene or simple vinylmation ether, 1,3,5-trihydroxybenzene-2,4,6-trisulfonic acid and carboxymethylcysteine acid, salts of various alkali metal, ammonium and substituted ammonium and polixeni acids, such as ethylenediaminetetraacetic acid, nitrilotriacetic acid, and polycarboxylate, such as malletova acid, succinic acid, citric acid, accidentally acid, primulina acid, benzene-1,3,5-tricarboxylic acid, carboxymethylcysteine acid and its soluble salts.

In some embodiments, implementation of the present invention the composition contains from about 0 to about 50 wt.% one or more structure-forming components selected from the group consisting of alkali metal salts and alkanolamine salts of the following compounds: phosphates, phosphonates, phosphonocarboxylates, salts of amino acids, electrolytes in the form of macromolecular aminopolyamide, medicationbuy polymers, salts of dicarboxylic acids and aluminosilicate salts. Examples of suitable divalent connecting metal ion in the chelate complex as described in British patent application No. 2094826 A, the description of which is included in this description by reference

In additional embodiments of the invention the compositions of this invention contain as alkalis or inorganic electrolytes from about 1 to about 50 wt.%, preferably from about 5 to about 30 wt.%, based on the weight of the composition, of one or more alkali metal salts of the following compounds: silicates, carbonates and sulfates, and organic bases such as triethanolamine, diethanolamine, monoethanolamine, triisopropanolamine.

In yet some additional embodiments, the implementation of the present invention the compositions contain as an anti-re-deposition from about 0.1 to about 5 wt.% one or more of the following compounds: polyethylene glycol, polyvinyl alcohol, polyvinylpyrrolidone and carboxymethylcellulose. In some preferred versions of the invention, the concatenation is used carboxymethylcellulose and/or polyethylene glycol with a composition of the present invention as useful graziadei compositions.

In some additional embodiments, the implementation of the present invention the bleaching agent(a), such as percarbonate sodium, perborate sodium adduct of sodium sulfate/hydrogen peroxide, and the adduct of sodium chloride/hydrogen peroxide is/or photosensitive bleaching dye, such as zinc or aluminum salt of sulfonated phthalocyanine, additionally increases detergents (detergent) exposure cleaning/bleaching compositions of the present invention. In additional embodiments of the invention are used in the bleaching accelerators (for example, TAED and/or NOBS).

In some embodiments, implementation of the present invention in the composition is included podsinwowa funds and/or fluorescent dyes. Examples of suitable podsinwowa funds and fluorescent dyes described in British patent application No. 2094826 A, the description of which is included in this description by reference.

In some embodiments, implementation of the present invention, in which the composition is a powder or solid composition included anti-caking agents. Examples of suitable anti-caking agents include saltsp-toluensulfonate acids, salts cellsurvival acid, salts of acetic acid, salt sulfonterol acid, talc, finely powdered silica, clay, calcium silicate (for example, Mikro-Cell from the company Johns Manville Co.), calcium carbonate and magnesium oxide.

In some embodiments, the implementation of the present invention find application antioxidants, including, but not limited to,tert-equivalent, 4,4'-butylidene the IP(6- tert-butyl-3-METHYLPHENOL), 2,2'-butylidene(6-tert-butyl-4-METHYLPHENOL), monotropaceae the cresol, Dictyostelium the cresol, monotropaceae phenol, Dictyostelium and 1,1-bis-(4-hydroxyphenyl)cyclohexane.

In yet some additional embodiments of the invention the compositions of the present invention also include soljubilizatory, including, but not limited to, lower alcohols (e.g. ethanol, salts benzosulfimide and salt lower alkylbenzenesulfonates, such as saltp-toluensulfonate), glycols such as propylene glycol, salt acetylbenzenesulfonyl, acetamide, amides pyridinedicarboxylic acid, salts benzoate and urea.

In some embodiments of the invention detergent compositions of the present invention are used in a wide range of pH, from acidic to alkaline pH. In a preferred embodiment of the invention the detergent composition of the present invention is used in moderately acidic, neutral or alkaline scrubbing medium with detergent having a pH of from greater than 4 to about not more than 12.

In addition to the above ingredients in the present invention also find use perfumes, buffers, preservatives, dyes and other Such components are used in concentrations and forms, known to experts in the Noi area.

In some embodiments of the invention fundamentals of powder detergents of the present invention receive any of the known methods of preparation, including the method of spray drying and/or granulation. In particular, the base detergent obtained by the method of spray drying and/or granulation with the spray drying is preferred. The base detergent obtained by the spray drying method is not limited on the terms of the receipt. The base detergent obtained by the spray drying method, is a hollow granules, which is produced by sputtering in the hot space aqueous suspension of heat-resistant ingredients, such as surface-active products and builders. After spray drying, you can add fragrances, enzymes, bleaching agents, inorganic alkaline builders. When using very dense, granular bases detergent obtained in this way, as the granulation with the spray drying, the various ingredients can also be added after getting the basics.

When detergent is a liquid, in some embodiments of the invention it is a homogenous solution, while in some and alternativnyj embodiments of the invention it is a non-homogeneous variance.

In some preferred versions of the invention, the detergent compositions according to the present invention is incubated with the tissue (for example, soiled tissues), industrial applications and applying for home use, temperatures, reaction times, and the modules baths, traditionally used in these environments. Specialists in this field will easily determine the conditions of curing (that is, the conditions effective for the treatment of materials by using detergent compositions according to the present invention).

As indicated above, in some embodiments, implementation of the present invention detergents get into pre-soaked in the appropriate solution form at intermediate pH, where there is sufficient activity to provide the required improvements in the bating, depilling, prevent peeling, removal and/or cleaning the surface of the fibers. In some embodiments, the invention also applies at least one surface-active substance. The remainder of the composition contains conventional components used in the solution for pre-soaking (for example, solvents, buffers, other enzymes (proteases), and so on) in their traditional concentrations.

In some embodiments of the invention purifying whom is osili of the present invention find application in the wash, cleaning hard surfaces, use in automatic dishwashing, as well as in cosmetic applications such as cleaning of dentures, teeth, hair and skin. The enzymes of the present invention also find application in other products for cleaning. Additional product in its simplest form can be a single or multiple stabilized enzymes of the present invention. This Supplement can be packaged in a unit dosage form for adding during the cleaning process. Standard dosage forms include, but are not limited to, briquettes, tablets, gel capsules or other standard dosage forms, such as pre-measured powders or liquids. In some embodiments of the invention to increase the amount of such compositions included the filling material and/or media. Suitable filler materials or media include, but are not limited to, various salts of sulfates, carbonates and silicates, as well as talc, clay or other filler Materials or media for liquid composition may be a water or a low molecular weight primary and secondary alcohols, including polyols and diols. Examples of such alcohols include, but are not limited to, methanol, ethanol, propanol and isopropanol. In some embodiments, implemented the I of the invention the compositions contain from about 5% to about 90% of these materials. In some alternative embodiments of the invention apply acid fillers to reduce the pH.

Cleaning compositions and cleaning additives of the present invention require effective amount of stabilized enzymes of the present invention. Usually cleansing compositions of the present invention contain at least of 0.0001 wt.%, from about 0.0001 to about 1, from about 0.001 to about 0.5, or even from about 0.01 to about 0.1 wt.% at least one enzyme of the present invention.

In some embodiments of the invention the cleaning composition of the present invention contain a material selected from the group consisting of a source of peroxide oxygen, hydrogen peroxide and mixtures thereof, the said source of peroxide oxygen selected from the group consisting of:

(i) from about 0.01 to about 50, from about 0.1 to about 20, or even from about 1 to 10 wt.% percale, organic peroxyacids, compounds of urea with hydrogen peroxide and mixtures thereof;

(ii) from about 0.01 to about 50, from about 0.1 to about 20, or even from about 1 to 10 wt.% carbohydrate and from about 0.0001 to about 1, from about 0.001 to priblizitel is but to 0.5, from about 0.01 to about 0.1 wt.% the carbohydrate oxidase and

(iii) mixtures thereof.

In some embodiments of the invention suitable parsoli include salt selected from the group consisting of alkali metal perborate, percarbonate alkali metal, perphosphate alkali metals, persulfates alkali metals and mixtures thereof.

In some preferred versions of the invention, the carbohydrate is selected from the group consisting of monoglide, diplomado, triplewides, oligoplites and mixtures thereof. Suitable carbohydrates include carbohydrates selected from the group consisting of D-arabinose, L - arabinose, D-cellobiose, 2-deoxy-D-galactose, 2-deoxy-D-ribose, D-fructose, L-fucose, D-galactose, D-glucose, D-glycero-D-Golovaty, D-lactose, D-lyxose, L-lyxose, D-maltose, D-mannose, melezitose, L-melibiose, palatinose, D-raffinose, L-ramnose, D-ribose, L-sorbose, stachyose, sucrose, D-trehalose, D-xylose, L-xylose, and mixtures thereof.

In some embodiments of the invention suitable carbohydrate oxidase includes a carbohydrate oxidase selected from the group consisting of alsoeasy (according to the IUPAC classification EC 1.1.3.9), galactosidase (according to the IUPAC classification EC 1.1.3.9), cellobioside (according to the IUPAC classification EC 1.1.3.25), pianosounds (according to the IUPAC classification EC 1.1.3.10), carbothioamides (according to the IUPAC classification EC 1.1.3.11, exotoxicity (according to the IUPAC classification EC 1.1.3.5) and/or glucose oxidase (IUPAC classification EC 1.1.3.4) and mixtures thereof.

In some alternative embodiments the invention, the cleaning compositions of the present invention also include from about 0.01 to about 99,9, from about 0.01 to about 50, from about 0.1 to about 20, or even from about 1 to about 15 wt.% molecules containing ester fragment. Suitable molecules that contain ester fragment include, but are not limited to, Poliplast, which contain the ester fragment. It is believed that any suitable ester fragment will be used in the present invention.

In some preferred embodiments of the invention proposed by the present invention the cleaning composition generally should be produced in such a way that during use in cleaning processes in the aquatic environment water for washing had a pH of approximately 5.0 to approximately 11,5 or even from about 7.5 to about 10.5. the Preparations in the form of liquid products usually get that way, so they have a pH from about 3.0 to about 9.0 in. Granular products for washing usually get that way, so they have a pH from about 9 to arr is siteline 11. Methods of pH control applied to the recommended levels include the use of buffers, alkalis, acids, etc. and is well known to specialists in this field.

In some embodiments of the invention, when the enzyme(s) of the present invention is used in granular or liquid compositions, it is desirable that the enzyme(s) was in the form of an encapsulated particle to protect this enzyme from other components of the granular composition during storage. In addition, encapsulation is also a means of controlling the availability of the enzyme(s) during the cleaning process and may enhance the characteristics of the enzyme(s). It is believed that this invention will find application in any suitable kapsulirujushchej material. In kapsulirujushchej material typically encapsulates at least part of the enzyme(s). Usually kapsulirujushchej material is water-soluble and/or dispersible in water. Moreover, it is believed that the cleansing composition of the present invention must be in any suitable form and be able to use any method chosen by the developer formulations (see, for example, U.S. patents№№ 5879584, 5691297, 5574005, 5569645, 5565422, 5516448, 5489392 and 5486303; all of which are included in this description by reference to non-limiting examples).

Some, especially repectfully embodiments of the invention proposed by the present invention the cleaning compositions find use in the purification in situ(for example, on the surface of the fabric or hard surface). Typically at least part of the seat in contact with the embodiment proposed by the present invention the cleaning composition in pure form or diluted in the washing solution form, and then these places are optionally washed and/or rinsed. From the point of view of the purposes of the present invention washable includes, but is not limited to, cleaning and mechanical stirring. It is believed that the fabric includes any suitable fabric that can be washed in the real conditions applied by the consumer. Described cleaning composition is usually used in concentrations of from about 500 h/m to about 15000 hours per million in solution. When the solvent in the wash is water, the water temperature is usually in the range from approximately 5°to approximately 90°C, and when the designated washing include fabric, the mass ratio of water to fabric typically is from about 1:1 to about 30:1.

Experimental part

The following examples are provided to demonstrate and further illustrate certain preferred embodiments of the present invention and its aspects, and should not be interpreted as limiting its scope.

In the experimental part describes the I, which follows, use the following abbreviations: °C (degrees centigrade); rpm (revolutions per minute); H2O (water); HCl (hydrochloric acid); aa (amino acid); BP (a pair of nucleotides); TPN (thousand base pairs); KD (kilodaltons); g (grams); μg (micrograms); mg (milligrams); ng (nanogram); μl (Microlitre); ml (milliliters); mm (millimeters); nm (nanometers); μm (micrometer); M (molar); mm (millimolar); μm (micromolar); U (units); (volts); M (molecular weight); sec (seconds); min(s) (minute/minutes); h(s) (hour/hours); MgCl2(magnesium chloride); NaCl (sodium chloride); OD280(optical density at 280 nm); OD600(optical density at 600 nm); EtOH (ethanol); PBS (phosphate buffered saline [150 mm NaCl, 10 mm nutrifaster buffer, pH of 7.2]); SDS (sodium dodecyl sulphate); Tris (Tris(hydroxymethyl)aminomethane); TAED (N,N,N, N'-tetraacetylethylenediamine); wt./about. (weight to volume);./about. (volume to volume); GOX and GOx (glucose oxidase); AOX and AOx (alcohol oxidase); COX and Cox (cholinesterase); HOX and HOx (hexosaminidase); SOX and Sox (corbiculidae); AATCC (American Association of certified dyes for fabrics); WFK (wfk Testgewebe GmbH, Bruggen-Bracht, Germany); TestFabric (TestFabric Inc., Pittston, PA); Warwick Equest (Warwick Equest Ltd., Warwick International, Flintshire, UK); ATCC (American Type Culture Collection, Manassas, VA); Baker (J.T.Baker, Phillipsburg, NJ); NAEF (NAEF, Press and Dies, Inc., Bolton Landing, NY); Sigma (Sigma-Aldrich Chemical Co., St. Louis, MO) and Minolta (Konica Minolta. Glen Coe NY).

Example 1

Stabilization of glucose oxidase in the standard AATCC detergent in the presence of glucose and hydrosulfite sodium

This example describes experiments conducted to evaluate the stabilization of glucose oxidase in the presence of its substrate (i.e. glucose) and inhibitor (i.e hydrosulfite sodium). In these experiments we applied a standard AATCC detergent (liquid detergent American Association of Textile Chemists and Colorists Heavy Duty Liquid detergent, version 2003, without optical Brightener; key components include linear alkanesulfonyl, alcohol ethoxylate, propandiol, citric acid, fatty acid, caustic soda and water; purchased from the company TestFabrics).

In these experiments as a positive control sample used 100 mm Tris (pH 8,3) of 0.005% surfactant TWEEN®-100. The use of a pH of 8.3 was based on measuring the pH of AATCC detergent. Weighed three dvuhmillimetrovy tubes 0,990 g of AATCC detergent (series # 01282004) and three other tubes 0,990 g control buffer. Then all tubes were added 90 mg (500 mm) glucose substrate. Then to each tube was added 100, 50 and 10 mm hydrosulfite sodium (MM 106,1), respectively, including tubes with control buffer and control AATCC detergent. All tubes on hour was placed on the company of the ion plate, to ensure good mixing and solubilization of glucose in detergent. Then in six 2-ml tubes (three with control buffer containing glucose/bisulphite, and three tubes with AATCC detergent containing glucose/bisulfite) was added to 500 hours/million (0.5 mg, 14,88 µl) glucose oxidase (OXYGOTML-5000, a total of 5,379 U/ml; 33.6 mg/ml; Genencor). Then all the tubes were mounted on a rotating plate (60 rpm) at room temperature. The production of hydrogen peroxide was measured using rapid tests (peroxidase/ABTS - Baker Testrips; Baker) at different moments of time t=0+minutes, 12 minutes and 30 minutes. In a test tube containing 100 mm of bisulfite with glucose, the glucose oxidase in the liquid detergent, optionally controlled the premature formation of hydrogen peroxide during the additional period of time of 1 hour, 12 hours, 7 days, 12 days and up to 21 days after the start. The results obtained in these experiments are listed in table 1 (see example 2).

At time 0+ control buffer mixture containing 10 mm bisulfite inhibitor, developed >1 hour/million H2O2while the control buffer mixture containing 50 or 100 mm bisulfite, never worked out of the hydrogen peroxide during different time periods tested (see table 1).

At time 0+ detergent mixtures containing 10 mm or 50 mm byself the private inhibitor, developed >10 hours/million H2O2while the control buffer mixture containing 100 mm bisulfite, during the test period was not developed hydrogen peroxide (see table 1). The obtained results show that at a concentration of 100 mm bisulfite avoid the formation of hydrogen peroxide in the liquid detergent containing 500 mm glucose and 500 hours/million oxidase due to inhibition of the oxidase. Moreover, in the liquid detergent containing 100 mm hydrosulfite sodium, 500 mm glucose and 500 h/ml glucose oxidase, premature formation of hydrogen peroxide was not observed for more than 21 days.

Example 2

The formation of hydrogen peroxide by glucose oxidase in the wash in the washing solution in the presence of hydrosulfite sodium

This example describes experiments conducted to evaluate the formation of hydrogen peroxide by glucose oxidase in the presence of hydrosulfite sodium when washing in the washing solution. As in example 1, in these experiments we applied a standard AATCC detergent.

In these experiments as a positive control sample used 100 mm Tris (pH 8,3) of 0.005% surfactant TWEEN®-100. The choice of pH of 8.3 was based on measuring the pH of AATCC detergent. Weighed three dvuhmillimetrovy the wading and, containing 0,990 g of AATCC detergent (series # 01282004), and three other tubes 0,990 g control buffer. Then to each tube was added 90 mg (500 mm) glucose substrate. Then to each tube was added 100, 50 and 10 mm hydrosulfite sodium (MM 106,1) (reversible inhibitor), respectively (i.e., either in tubes with control buffer, and in tubes containing detergent AATCC). All tubes on hour were placed on a rotating plate to ensure good mixing and solubilization of glucose in detergent. Then cooked 6 tubes containing five ml of water for washing (5 mm HEPES, stiffness 6 grams per gallon, pH 8). Then in six 2-ml tubes (three with control buffer containing glucose/bisulphite, and three with AATCC detergent containing glucose/bisulfite) was added to 500 hours/million (0.5 mg, 14,88 µl) glucose oxidase (OXYGOTML-5000, a total of 5,379 U/ml; 33.6 mg/ml; Genencor). Then all the tubes were mounted on a rotating plate (60 rpm) at room temperature. The production of H2O2measured using rapid tests (peroxidase/ABTS)as described in example 1, for t=0+ minutes, 12 minutes and 30 minutes. Immediately after adding the enzyme to the cleaning means and the control mixture was extracted 10 ál of the mixture and was mixed with 5 ml of water for washing. Then all tubes were also tested for the presence of hydrogen peroxide, using expre the C-sample when the duration of the experiment 12 and 30 minutes. In the presence of 5 ml of washing solution final concentration of the enzyme glucose oxidase was 1 hour/million, and the glucose concentration was 1 mm.

The results indicated that the washing solution, containing control buffer was developed approximately 10 hours per million of hydrogen peroxide in the presence of the drug containing 10 mm bisulfite inhibitor, ~10 hours/mn in the presence of 50 mm bisulfite and ~3 hours/million H2O2in the presence of 100 mm bisulphite when the duration of the experiment 12 minutes. Thus, the results indicate reversible bisulfite inhibitors (for more details see the table below).

The washing solution containing detergent AATCC, when the duration of the experiment 12 minutes developed >3 hours/million for all three concentrations of bisulfite inhibitor, also proving reversible bisulfite inhibitor. In addition, the cleaning solution with detergent AATCC when the duration of the experiment 30 minutes produced ~10 hours/million for all three concentrations of bisulfite inhibitor, again proving reversible bisulfite inhibitor. The results show that sodium bisulfite is a reversible inhibitor, suitable for saving the glucose oxidase, ribereau in the presence of high concentrations of substrate. However, at a dilution of detergent in the water for washing the inhibition disappears. It is important to note that the sodium bisulfite is a reversible inhibitor of the glucose oxidase-dependent concentration.

Table 1
Determination of the formation of hydrogen peroxide (mg/l h/million)
Time (min)Control buffer with glucose oxidase hours/million N2About2Detergent AATCC with glucose oxidase hours/million N2About2
T=0+ in the case of 500 mm glucose, 500 hours/million oxidase and 10 mm bisulfite1>10
T=0+ in the case of 500 mm glucose, 500 hours/million oxidase and 50 mm bisulfite010
T=0+ in the case of 500 mm glucose, 500 hours/million oxidase and 100 mm bisulfite00
T=12, the cleaning solution, 1 mm glucose and 1 h/million oxidase33
T=12, the cleaning solution, 1 mm glucose, 1 hour/million oxidase and 0.02 mm bisulfite ~103
T=12, the cleaning solution, 1 mm glucose, 1 hour/million oxidase and 0.1 mm bisulfite33
T=12, the cleaning solution, 1 mm glucose, 1 hour/million oxidase and 0.2 mm bisulfite~33
T=30, the cleaning solution, 1 mm glucose, 1 hour/million oxidase and 0.02 mm bisulfite1010
T=30, the cleaning solution, 1 mm glucose, 1 hour/million oxidase and 0.1 mm bisulfite~10~10
T=12, the cleaning solution, 1 mm glucose, 1 hour/million oxidase and 0.2 mm bisulfite~3~10
T=12, the cleaning solution, 1 mm glucose and 1 h/million oxidase1010

Example 3

Stabilization of glucose oxidase in detergent containing glucose and sodium metabisulfite

This example describes experiments conducted to evaluate the formation of hydrogen peroxide by glucose oxidase in the presence of sodium metabisulfite (reversible inhibitor oxidase) when washing in the washing solution. As in examples 1 and 2, in these experiments used the standard AATCC detergent.

In these experiments we used 100 mm Tris (pH 8,3) of 0.005% surfactant TWEEN®-100 as a positive control sample. As described above, the choice of a pH of 8.3 is based on measuring the pH of AATCC detergent. Weighed three dvuhmillimetrovy tubes 0,990 g of AATCC detergent (series # 01282004) and three other tubes 0,990 g control buffer.

Then to each tube was added 90 mg (500 mm) glucose substrate. Then was added 100, 50 and 10 mm sodium metabisulfite, respectively (i.e., either in tubes containing control buffer or in tubes containing detergent AATCC). All tubes on hour were placed on a rotating plate to ensure good mixing and solubilization of glucose in detergent. Then in six 2-ml tubes (three with control buffer containing glucose/metabisulfite, and three with AATCC detergent containing glucose/metabisulfite) was added to 500 hours/million (0.5 mg, 14,88 µl) glucose oxidase (OXYGOTML-5000, a total of 5,379 U/ml; 33.6 mg/ml; Genencor). Then all the tubes were mounted on a rotating plate (60 rpm) at room temperature. The production of H2O2measured using rapid tests (peroxidase/ABTS)as described in examples 1 and 2, for the time of the experiment, t=0+minutes, 1 minute and 30 minutes. In a test tube containing 100 mm of bisulfite with glucose, the glucose oxidase in the liquid detergent, optionally controlled the premature formation of hydrogen peroxide during the additional period of time of 1 hour, 12 hours, 7 days, 12 days and up to 21 days after the start. The results are shown in the following table 2.

At time 0+ control buffer mixture containing 10 mm metabisulfite inhibitor, was developed 1 h/million H2O2while the control buffer mixture containing 50 or 100 mm bisulfite, never worked out of the hydrogen peroxide during different time periods tested (see table 2).

At time 0+ detergent mixtures containing 10 mm or 50 mm metabisulfite inhibitor, developed >10 hours/million H2O2while the control buffer mixture containing 100 mm bisulfite, never worked out of the hydrogen peroxide within the time periods tested (see table 2). The obtained results show that at a concentration of 100 mm metabisulfite avoid the formation of hydrogen peroxide in the liquid detergent containing 500 mm glucose and 500 h/ml glucose oxidase, due to inhibition of the oxidase. Moreover, in the liquid detergent containing 100 mm metasulfite sodium, 500 mm glucose and 500 h/ml glucose oxidase, p is estavromenos the formation of hydrogen peroxide was not observed for more than 21 days.

Example 4

The formation of hydrogen peroxide by glucose oxidase in the washing solution for washing in the presence of sodium metabisulfite

This example describes experiments conducted to evaluate the formation of hydrogen peroxide by glucose oxidase in the presence of sodium metabisulfite (reversible inhibitor oxidase) when washing in the washing solution. As in the above examples, these experiments used the standard AATCC detergent.

In these experiments we used 100 mm Tris (pH 8,3) of 0.005% surfactant TWEEN®-100 as a positive control sample. The choice of pH of 8.3 is based on measuring the pH of AATCC detergent. Weighed three dvuhmillimetrovy tubes containing 0,990 g of AATCC detergent (series # 01282004), and three other tubes 0,990 g control buffer. Then to each tube was added 90 mg (500 mm) glucose substrate. Then in each tube (i.e., either in tubes containing control buffer, and tubes containing detergent AATCC) was added at 100, 50 and 10 mm sodium metabisulfite, respectively. All tubes on hour were placed on a rotating plate to ensure good mixing and solubilization of glucose in detergent. Then cooked 6 tubes containing five ml of water for washing (5 mm HEPES, stiffness 6 grams on the Mallon are optimistic about, pH 8). Then in six 2-ml tubes (three with control buffer containing glucose/bisulphite, and three with AATCC detergent containing glucose/bisulfite) was added to 500 hours/million (0.5 mg, 14,88 µl) glucose oxidase (OXYGOTML-5000, a total of 5,379 U/ml; 33.6 mg/ml; Genencor). Then all the tubes were mounted on a rotating plate (60 rpm) at room temperature. The production of H2O2measured using rapid tests (peroxidase/ABTS)as described in the above examples, for the time of the experiment, t =0+ minutes, 12 minutes and 30 minutes. Immediately after adding the enzyme to the cleaning means and the control mixtures were removed and 10 μl of the mixture was mixed with 5 ml of water for washing. Then all tubes were also tested for the presence of hydrogen peroxide, using rapid tests, when the duration of the experiment 12 and 30 minutes. In the presence of 5 ml of washing solution final concentration of the enzyme glucose oxidase was 1 hour/million, and the glucose concentration was 1 mm.

The results indicated that the washing solution, containing control buffer was developed approximately 10 hours/million H2O2in the presence of 10 mm metabisulfite inhibitor, ~10 hours/mn in the presence of 50 mm metabisulfite and ~3 hours/million H2O2in the presence of 100 mm metabisulfite when the duration of the experiment 12 minutes. Thus, obtained the results indicate reversible metabisulfite inhibitor (for more details see the chart below).

The washing solution containing detergent AATCC, developed >3 hours/mn in the presence of all three concentrations metabisulfite inhibitor when the duration of the experiment, 12 minutes, also proving reversible metabisulfite inhibitor. In addition, the cleaning solution with AATCC detergent was developed ~10 hours/mn in the presence of all three concentrations metabisulfite inhibitor when the duration of the experiment 30 minutes, again proving reversible metabisulfite inhibitor. After 3 weeks of storage in the presence of inhibitor at a dilution in washing liquid preparations of detergent worked out 3 hours per million of hydrogen peroxide for 12 minutes. To 30 minutes produces ~10 hours/million of hydrogen peroxide.

The results show that sodium metabisulfite is a reversible inhibitor suitable for conservation oxidase, inhibiting in the presence of high concentrations of substrate. However, at a dilution of detergent in the water for washing the inhibition disappears. It is important to note that sodium metabisulfite is a reversible inhibitor of the glucose oxidase-dependent concentration.

In addition to sodium metabisulfite and sodium bisulfite as described in the examples for the inhibition of oxidase felt the other is their inhibitors. Used methods similar to those described in the examples. The results showed that 1 M fluoride or sodium thiosulfate inhibit the oxidase in a small degree. However, it was determined that 2 M hydroxylamine can stabilize the premature formation of hydrogen peroxide in detergent containing 1 M glucose and 500 hours/million oxidase.

Table 2
Definition N2About2
Time (min)Control buffer (glucose oxidase) N2About2hours/millionDetergent AATCC (glucose oxidase) N2About2hours/million
T=0+, 12, 30 in the case of 500 mm glucose and 500 hours/million enzyme, 100 mm sodium metabisulfite00
T=12, the cleaning solution, 1 mm glucose, 1 h/ml glucose oxidase, without inhibitor33
T=12, the cleaning solution, 1 mm glucose and 1 h/million enzyme, 0.2 mm of sodium metabisulfite13
T=30, the cleaning solution, 1 is M glucose and 1 h/million enzyme 0.2 mm of sodium metabisulfite3~10
T=30, the cleaning solution, 1 mm glucose oxidase without inhibitor1010
T=60, the cleaning solution, 1 mm glucose, 1 h/ml glucose oxidase, 0.2 mm of sodium metabisulfite10>10

Example 5

Stabilized alcohol oxidase in detergent-containing substrate and inhibitor

This example describes experiments conducted to evaluate the stabilization of alcohol oxidase in the presence of its substrate (ethanol) and inhibitor (for example, metabisulfite, bisulfite or sodium thiosulfate). As in the above examples, these experiments used the standard AATCC detergent.

In these experiments tested the stability of alcohol oxidase obtained fromHansunela sp. (100 U/ml 22 U/mg, 13 mg total solids in the ampoule, 7,7 U/mg solid; Sigma), in the liquid detergent AATCC. In these experiments, each experiment used 10 Units of alcohol oxidase in the mother liquid detergent. Testing was stirred 1 M ethanol and detergent (46 mg of ethanol 990 mg of detergent). The presence of 1 M ethanol is e affect the General appearance of a liquid detergent. As reversible inhibitors of alcohol oxidase felt hydrosulfite sodium (NaHSO3), sodium metabisulfite (Na2S2O5) and sodium thiosulfate (Na2S2O3). The experiments were conducted as described above in examples 1 and 3.

It was found that the enzyme alcohol oxidase is stable in the liquid detergent AATCC over a period of time (120 minutes) test in the presence of ethanol as substrate and inhibitors. It was found that thiosulfate is a weak inhibitor of alcohol oxidase, while hydrosulfite and sodium metabisulfite are reversible inhibitors of alcohol oxidase test concentration of 100 mm. At concentrations of 100 mm hydrosulfite and sodium metabisulfite are able to stop premature formation of H2O2in the fallopian AATCC detergent solution during the study period (120 minutes), as shown in tables 3 and 4.

Example 6

Education H2O2alcohol oxidase in the washing solution for washing containing ethanol and reversible inhibitor

This example describes experiments conducted to evaluate the formation of hydrogen peroxide alcohol oxidase in the presence of metabisulfite, bisulfite and sodium thiosulfate (reversible inhibitors of alcohol oxidase) in washing the feet to the washing solution. As in the above examples, these experiments used the standard AATCC detergent.

The stability and activity of the enzyme alcohol oxidase (described in example 5) in the liquid detergent AATCC (Sigma, 100 U/ml 22 U/mg, 13 mg total solids in the ampoule, 7,7 U/mg solid) were tested at a dilution in the washing solution. In these experiments, for each experiment we used 10 Units of alcohol oxidase in the mother liquid detergent. In addition, 1 M ethanol (substrate) was mixed with detergent (46 mg of ethanol 990 mg of detergent). When breeding 500× wash solution contained 2 mm ethanol for education maximum concentration of 2 mm H2O2and a finite amount of alcohol oxidase in the washing solution, equal to 0.02 Units. As reversible inhibitors had hydrosulfite sodium (NaHSO3), sodium metabisulfite (Na2S2O5) and sodium thiosulfate (Na2S2O3). In these experiments, 5 ml of washing solution was added 10 μl of the final washing of the mixture or of the control mixture. In these experiments we used methods similar to those described in examples 2 and 4.

When breeding in the washing solution of hydrogen peroxide formed in the presence of detergent containing alcohol oxidase, ethanol and inhibitor (e.g., Hydra is sodium sulfite or sodium metabisulfite), as indicated in tables 3 and 4.

Table 3
Determining the concentration of hydrogen peroxide in the liquid detergent
Time (min)Control buffer (alcohol oxidase) N2About2hours/millionDetergent AATCC (alcohol oxidase) N2About2hours/million
T=0+ in the case of 1 M ethanol and 10 Units of the enzyme without inhibitor1010
T=0+ in the case of 1 M ethanol, 10 Units of the enzyme, 100 mm sodium metabisulfite (detergent condensing)00
T=0+ in the case of 1 M ethanol, 10 Units of the enzyme, 100 mm sodium thiosulfate~103
T=0+ in the case of 1 M ethanol, 10 Units of the enzyme, 100 mm hydrosulfite sodium00
T=12, in the case of 1 M ethanol and 10 Units of the enzyme without inhibitor3030
T=12 in the case of 1 M ethanol, 10 Units of the enzyme, 100 mm met the bisulphite sodium 00
T=12 in the case of 1 M ethanol, 10 Units of the enzyme, 100 mm hydrosulfite sodium00
T=30, in the case of 1 M ethanol and 10 Units of the enzyme without inhibitor3030
T=30 in the case of 1 M ethanol, 10 Units of the enzyme, 100 mm sodium metabisulfite00
T=30 in the case of 1 M ethanol, 10 Units of the enzyme, 100 mm hydrosulfite sodium00
T=120 in the case of 1 M ethanol and 10 Units of the enzyme without inhibitor3030
T=120 in the case of 1 M ethanol, 10 Units of the enzyme, 100 mm hydrosulfite, metabisulfite or sodium thiosulfate00 (for thiosulfate >10)

Table 4
Determining the concentration of hydrogen peroxide in the washing solution
Time (min)Control buffer (alcohol oxidase) N2 About2hours/millionDetergent AATCC (alcohol oxidase) N2About2hours/million
T=0+, the cleaning solution, 2 mm ethanol and 0.02 Units of enzyme without inhibitor0
T=0+, the cleaning solution, 2 mm ethanol, 0.02 Units of enzyme, 0.2 mm of sodium metabisulfite0
T=0+, the cleaning solution, 2 mm ethanol, 0.02 Units of enzyme, 0.2 mm sodium thiosulfate00
T=0+, the cleaning solution, 2 mm ethanol, 0.02 Units of enzyme, 0.2 mm of hydrosulfite sodium00
T=12, the cleaning solution, 2 mm ethanol and 0.02 Units of enzyme without inhibitor10
T=12, the cleaning solution, 2 mm ethanol, 0.02 Units of enzyme, 0.2 mm of sodium metabisulfite00
T=12, the cleaning solution, 2 mm ethanol, 0.02 Units of enzyme, 0.2 mm of hydrosulfite sodium00
T=30, the cleaning solution, 2 mm ethanol and 0.02 E is enzyme, without inhibitor00
T=30, the cleaning solution, 2 mm ethanol, 0.02 Units of enzyme, 0.2 mm of metabisulfite or hydrosulfite sodium00
T=30, the cleaning solution, 2 mm ethanol, 0.02 Units of enzyme, 0.2 mm sodium thiosulfate00
T=120, the cleaning solution, 2 mm ethanol and 0.02 Units of enzyme without inhibitor~10~10
T=120, the cleaning solution, 2 mm ethanol, 0.02 Units of enzyme, 0.2 mm of metabisulfite, hydrosulfite or sodium thiosulfate1 (hydrosulfite) 0 (metabisulphite)>3(hydrosulfite) 1(metabisulphite) >3 (thiosulfate)

In additional experiments also assessed the ability of 10 mm CuSO4to stabilize the premature formation of H2O2in detergent containing 1 M ethanol and 10 Units of alcohol oxidase (Candida sp.; Sigma).

Example 7

Stabilization cholinesterase in detergent containing choline and inhibitor

This example describes experiments conducted to evaluate the stabilization of cholinesterase in the presence of its substrate (i.e. Ho is ina) and inhibitor (i.e. sodium bisulfite and 2-amino-2-methyl-1-propanol). In these experiments used the standard AATCC detergent.

The experiments were conducted as in example 1. Cholinesterase produces two mol H2O2per mole of choline. The results obtained in these experiments prove that cholinesterase was stable in detergent AATCC over a 24-hour period of testing in the presence of choline and inhibitors. The inhibitor 2-amino-2-methyl-1-propanol (AMP) is a reversible inhibitor cholinesterase and stops premature restoration of H2O2in detergent, when used at a concentration of 200 mm. It was found that hydrosulfite sodium is a reversible inhibitor cholinesterase at the tested concentration of 100 mm. Hydrosulfite sodium at a concentration of 100 mm) is also able to stop premature formation of H2O2in the fallopian AATCC detergent solution. When breeding in the detergent solution detergent containing cholinesterase, choline chloride and inhibitor (e.g., hydrosulfite or sodium 2-amino-2 metal-1-propanol), over time produces H2O2as shown in table 5.

Table 5
Definition N2About2in the liquid detergent and the washing solution
Time (min)Control buffer (cholinesterase) N2About2hours/millionDetergent AATCC (cholinesterase) N2About2hours/million
T=0+, 1 M choline and 1 U of enzyme without inhibitor1010
T=0+, 1 M choline, 1 Unit of enzyme, 100 mm inhibitor AMR00
T=12, 1 M choline, 1 Unit of enzyme, 100 mm inhibitor AMR11
T=12, 1 M choline, 1 IU or 10 IU of the enzyme, 200 mm of inhibitor AMR(pH ~9,3)00
T=0+, 12, 30 and 60, the cleaning solution, 2 mm holdingarea, 0.02 Units of enzyme, 0.4 mm of inhibitor AMR1,3,3,101,3,3,10
T=0+, 12, 30 and 60, 1 M choline chloride, 10 Units of cholinesterase, 100 mm inhibitor of sodium bisulfite (pH 7)00
T=0+, 12, 30 and 60, the cleaning solution, 2 mm choline chloride, 0.02 Units of cholinesterase, 0.2 mm of inhibitor of sodium bisulfite (pH 7)1,10,>10.30 a.m. 1,10,>10.30 a.m.

Example 8

Stability/performance oxidase and exotoxicity in detergent containing sodium bisulfite and glucose

This example describes experiments conducted to assess the stability of glucose oxidase and exotoxicity in detergent containing sodium bisulfite and glucose. Also describes additional experiments to assess the characteristics of these enzymes in relation to contaminated tissue samples.

In four 250 ml glass bottles were mixed with 100 grams of liquid detergent AATCC with 9 grams of glucose and was stirred for 30 minutes to dissolve glucose in detergent. Then two bottles was added 2,12 grams of sodium bisulfite to be dissolved in detergent. Then in two bottles (one with bisulfite and without bisulfite) was added to 15000 units of glucose oxidase and similarly in the other two bottles (one with bisulfite and without bisulfite) was added to 15000 units exotoxicity. All four liquid preparation of detergent was stored at room temperature and evaluated their effectiveness in removing stains using disc-shaped tissue samples in a 12-hole tablets within 7 days.

Tissue samples contaminated with blueberries and tea (CS 15-004, CS3; TestFabric), was cut into 15 mm krugis by using a die press for fabrics (model 93046; NAEF), equipped with a 5/8" shancevym knife. Separate disks were placed in each well of a 24-hole tablet (Costar). To each well was added one (1) ml of washing solution (pH of 10.0)containing per liter of 1.5 ml of detergent AATCC HDL, 10 mm sodium carbonate, 75 mm glucose, with a hardness of 6 grams per gallon (obtained by diluting the mother liquor with rigidity 15000 grams per gallon, containing 1,735M of calcium chloride and 0.67 M of magnesium chloride) and 0.05% TAED (tetraacetylethylenediamine, Fluka). In 4 wells of one section with a pipette and preemptive actions added five (5) microliters glucose oxidase obtained 5-7 days ago, with sodium bisulfite or without sodium bisulfite. Control wells (8) did not contain enzyme.

The microplate was covered with its plastic lid and kept at 37°C and a smooth rotation with a speed of 100 rpm After 5 hours the supernatant liquid was removed by aspiration and each well washed twice with 1.5 ml of phosphate buffered saline, Dulbecco (Dulbecco PBS, pH 7.3) and twice with 1.5 ml of distilled water. Each disk was removed from his hole and was dried overnight in air.

The discs were visually inspected and analyzed using reflectometer Minolta Reflectometer CR-200, calibrated by a standard white tile. Expected average values of L. the light reflectance of the surface of the Kani was measured as Lambertucci the reflection coefficient, called "L-factor" (the ratio of reflected to incident light on the surface of light, usually expressed in percent), on the surface of the material such a large thickness that the reflection does not change with increasing thickness (i.e., the reflection coefficient of the surface), regardless of other parameters, such as the reflectance of the rear surface. L-factor was measured by measuring the reflection using the above Refractometer, as a percent stain removal (%SR=100%×(final reflectance initial reflectance)/(reflectance of white standard-initial reflectance)).

After 5 days oxidase, prepared without bisulfite, became yellow, contained a large number (>30 mg/l) of hydrogen peroxide in the obtained sample showed minimal activity on the production of hydrogen peroxide (1 mg/l) during the test drive and had the bleaching characteristics, which were not statistically different from the control sample without enzyme. In contrast, after 7 days oxidase, prepared bisulfite was white, showed active (>100 mg/l) production of hydrogen peroxide during the test drive and showed a performance significantly better than the control is on the sample, and glucose oxidase without bisulfite. The same results were observed after 14 days. The results are shown in table 6 and 7 and figures 1 and 2. The results show that glucose oxidase, stable bisulfite, whitens disks, contaminated blueberries, and drives contaminated tea, significantly better than the control sample (without enzyme) or non-stabilized glucose oxidase.

69,42
Table 6
Operational characteristics of the liquid detergent containing oxidase, in relation to removing blueberry stains on day 5 for non-stabilized glucose oxidase and at day 7 for oxidase, stable bisulfite
5 days7 days
without glucose oxidaseglucose oxidaseglucose oxidase, stable bisulfite
% stain removal (dL)17,218,823,2
L-factor68,9370,74
Standard deviation0,330,320,42

Table 7
Operational characteristics of the liquid detergent containing oxidase, in relation to the removal of tea stains on day 14 for non-stabilized glucose oxidase and on day 16 for oxidase, stable bisulfite
14 days16 days
without glucose oxidaseglucose oxidaseglucose oxidase, stable bisulfite
% stain removal (dL)6,91232,2
L-factor75,1476,2an 80.2
Standard deviation0,370,360,35

<> Example 9

Test ergotamine on stability/performance oxidase in detergent containing sodium bisulfite and glucose, within two months

This example describes experiments conducted to assess the stability of glucose oxidase (GOX) in the liquid detergent containing sodium bisulfite and glucose. Also describes additional experiments to evaluate the performance characteristics of these enzymes in tissue samples with numerous dirt.

In two 250 ml glass bottles were mixed with 100 grams of liquid detergent AATCC with 9 grams of glucose and was stirred for 30 minutes to dissolve glucose in detergent. Then in one of the bottles was added 2,12 grams of sodium bisulfite (Sigma Aldrich # 243973) and was dissolved in detergent. Then in both bottles (bottle bisulfite and into the bottle without bisulfite) was added 15000 Units of glucose oxidase (HPL5000, total of 5,379 U/ml; Genencor). Both the liquid preparation of detergent was stored at room temperature for two months and were evaluated for their efficacy to remove stains, using tissue samples from numerous pollution (Warwick Equest) ergotamine.

When testing in ergotamine in the vessel 1 and 4 were added to 950 ml of water MilliQ Water and 950 ml of MilliQ Water was added to the vessel 2, and 3. In the vessel 1 and added fifty ml of a 1.5 M solution of glucose. In the vessel 1 and 4 were added three ml of fresh AATCC detergent, whereas in the vessel 2, and 3 were cleanser ATCC two months ago with GoX (i.e. without bisulfite and bisulfite), as described above. The final concentration of 2 mm bisulfite taught in the vessel 1 and 4 by adding a stock solution of sodium bisulfite (1 M). The hardness of the water was maintained at a level of 6 grams per gallon (i.e. washing conditions in North America). The final concentration of TAED all vessels remained at the level of 0.05% along with the addition of sodium carbonate to bring the pH to a value in the range from 8.5 to 9.15. Pre-prepared twenty-four sample of tissue with numerous contaminants in each vessel was added six tissue samples and stirred at 125 rpm and Then into the vessel 4 was added to 100 MK stock solution of glucose oxidase, whereas in the vessel 1 glucose oxidase was added. Thus, the vessels 1 and 4, respectively, were used as negative and positive control samples. The experiment was carried out for 90 minutes at 30°C. After the test in ergotamine tissue samples were washed (3×) with cold tap water, squeezed and then dried over night at room temperature. All tissue samples were pressed under steam and then evaluated using the reflectometer Minolta.

Research in ergotamine prove with ailnoth GOX, obtained together with bisulfite, and also prove the advantage of its bleaching properties compared to GOX without bisulfite and a control sample against stains from coffee, wine, Merlot, blueberries, black currants and mix berries (see table 8 and figure 3).

All patents and publications cited in the specification are indicative of the level of technology for professionals in this field, which relates to the invention. All patents and publications incorporated in this description by reference to the same extent as if each individual publication was specifically and individually indicated to be incorporated by reference.

Having described the preferred embodiments of the present invention, the person skilled in the art it will be obvious that the described embodiments can be obtained various modifications and such modifications shall be included within the scope of the present invention.

Specialists in this field it is easy to understand that the present invention is well adapted to carry out the objects and to attain the ultimate goals and the advantages mentioned, as well as its inherent advantages. Described in this specification, compositions and methods are preferred embodiments of the invention, t is Auda illustrative and are not considered limiting the scope of invention. The person skilled in the art it will be obvious that it is possible to make various changes and modifications of the presented invention without going beyond the scope and essence of the invention.

Illustrative presents the invention in a suitable manner can be realized in practice in the absence of any element or elements, limitation or limitations which is not specifically described in this description. The terms and expressions that are used are used as terms of description and are not limiting, and there is no intention in the use of such terms and expressions to exclude any equivalents of the above described signs or portions thereof, but it is believed that within the scope of the claimed invention, various modifications are possible. Thus, it should be understood that although the invention is specifically described using preferred embodiments and optional features, modification and variation described in this description of the concepts, you can enlist the help of specialists in this area, and that such modifications and variations should considered within the scope of the present invention, as defined by the attached claims.

The invention is described in broad and General sense. Each of the narrower species characteristics and subgroups in the General description, which is also part of the invention. It covers a General description of the invention with the proviso or negative limitation excluding any object of study of the class of objects, regardless of whether there is in this description specific reference to excluded material or not.

1. Stabilized oxidase composition containing the specified oxidase, and a stabilizer, and at least one substrate of the specified oxidase, where the specified stabilizer contains at least one oxidase inhibitor, and the specified stabilizer selected from thiosulfate and 2-amino-2-methyl-1-propanol.

2. The composition according to claim 1, where the specified oxidase selected from glucose oxidase, corbiculidae, cholinesterase, exotoxicity and alcohol oxidase.

3. The composition according to claim 1, where the specified substrate is chosen from glucose, lactate, sorbitol, choline, glycerol, ethylene glycol, propylene glycol and ethanol.

4. The composition according to claim 1, where the specified stabilizer contains at least one sulfite.

5. The composition according to claim 4, where the specified at least one sulfite selected from hydrosulfite sodium, metabisulfite, sodium and/or sodium bisulfite.

6. The composition according to claim 1, where the specified composition is a cleansing, whitening or disinfectant composition.

7. The composition according to claim 6, where the specified detergent is a detergent for washing is whether detergent for washing dishes.

8. The composition according to claim 7, where the specified detergent selected from a powder, liquid and gel detergents.

9. The composition according to claim 1, where the specified composition is a detergent (detergent) Supplement or product pre-processing.

10. The composition according to claim 1, additionally containing activator bleaching or bleach precursor.

11. The composition of claim 10, where the specified activator selected from precursors of percolate, metal complexes, peroxidase and systems acyltransferase/substrate.

12. The composition according to claim 1, additionally containing at least one enzyme selected from proteases, amylases, pectinase, pectacles, lipases, mannanase, cellulases, esterases, koutinas, oxidoreductases, hemicellulase and karbohidrat.

13. The composition according to claim 1, additionally containing at least one auxiliary ingredient chosen from surfactants, builders, bleaching products, antimicrobial agents, polymers, solvents, salts, buffer funds, chelat forming means, means inhibiting the transfer of dyes, additives for deposition, dispersing agents, enzymes, enzyme stabilizers, catalytic materials, bleach activators, accelerators whitening, previously obtained perkiset, polymer disperser the General funds graziadei tools/anti re-deposition, optical brighteners, soap suds suppressors, dyes, perfumes, tools, giving elasticity to the structure, fabric softeners, carriers, hydrotropes, technological additives, pigments and mixtures thereof.

14. The method of formation of the whitening product in the washing solution, including the stage of adding the specified composition according to claim 1 to the specified cleaning solution.

15. The method according to 14, where specified whitening product is a peroxide or whitening system that can be activated using peroxide.



 

Same patents:

FIELD: chemistry.

SUBSTANCE: invention relates to a biocidal composition containing hydrogen peroxide in concentration of 0.05-50% (wt/wt) and a compound of formula 1: (OH)(2-m)(X)(O)P-[(O)p-(R')q-(CH(Y)-CH2-O)n-R]m, where X is H or OH; each Y is independently H or CH3; m equals 1 and/or 2; each p and q is independently equal to 0 or 1, provided that if p equals 0, q equals 1; each n is independently equal to 2-10; each R' is independently an alkylene radical containing 1-18 carbon atoms; each R is independently H or an alkyl radical containing 1-18 carbon atoms; and R'+R≤20; in concentration of 0.01-60% (wt/wt), as a biocidal composition. The invention also relates to use of the disclosed composition as a biocidal composition, as well as for purposes where there is need for disinfection and/or sanitation activity.

EFFECT: composition has excellent biocidal activity.

25 cl, 9 tbl, 10 ex

FIELD: chemistry.

SUBSTANCE: aqueous composition has pH equal to or less than 3, and contains the following ingredients (per total weight of the composition) : a) 0.05-20 wt % polymer thickener containing 20-100000 monomer links and an average of at least 0.8-COOR groups per monomer link, where for each separate -COOR group, R is independently selected from H, OH, and a carbon-containing group, b) 0.05-30 wt % hydrogen peroxide in terms of H2O2, c) 0.5-60 wt % one or more aliphatic carboxylic acids containing 1-8 carbon atoms, their alkyl esters, anhydrides and/or peroxy acids. Content of active oxygen, which is ensured by presence of ingredients a) and c) is equal to at least 0.02 wt % (per total weight of the composition).

EFFECT: obtaining a stable aqueous composition with low pH for cleaning different surfaces and for bleaching textile materials or paper.

15 cl, 10 ex, 3 tbl

FIELD: chemistry.

SUBSTANCE: sodium percarbonate particles covered by a cladding have a core of sodium percarbonate obtained via fluidised bed granulation. The cladding contains sodium sulphate and sodium carbonate in weight ratio ranging from 95:5 to 75:25 in relative amount of at least 80 wt %.

EFFECT: improved detergent action of sodium percarbonate when used as detergent component with simultaneous increase in stability when stored with detergents.

12 cl, 5 tbl

FIELD: chemistry.

SUBSTANCE: sodium percarbonate particles are covered by a cladding which contains sodium sulphate in form of a high-temperature phase of sodium sulphate and/or in form of a high-temperature phase of a double salt of formula Na4(SO4)1+n(CO3)1-n, where n is a number ranging from 0 to 0.5.

EFFECT: high stability during storage.

8 cl, 4 tbl, 3 ex

FIELD: chemistry.

SUBSTANCE: sodium percarbonate particles are covered by a cladding which contains anhydrous sodium sulphate in amount of 70-99.8 wt % and sodium borate in amount of 0.2-20 wt % and where said cladding accounts for 1-10% of the total weight of one particle of sodium percarbonate.

EFFECT: high stability during storage when using sodium percarbonate particles as a component of detergent and cleaning agents.

9 cl, 2 tbl, 2 ex

FIELD: chemistry.

SUBSTANCE: invention relates to a bleaching system for household textile items containing at least one bleaching agent, where the bleaching system is selected from peroxybenzoic acid, peroxy-6-naphthoic acid, peroxylauric acid, peroxystearic acid, phthalimido peroxycaproic acid, 6-phthalimido peroxyhexanoic acid, nonylimido peroxyamber acid, nonylimido peroxyadipic acid, 1,12-diperoxydodecanoic acid, 1,9-diperoxyazelaic acid, diperoxyisophthalic acid and 2-decyldiperoxybutane-1,4-diacid and coated by a shell in form of a layer of a polymer with urethane and urea groups, where a prepolymer with terminal NCO groups is obtained from macrools, ionic or potentially ionic polyols and polyisocyanates used in excess, said prepolymer being subjected to reaction with compounds which contain at least two amine groups which are reactive towards isocyanate with ratio of NCO groups to NH groups less than or equal to 1:1, after which said polymer is obtained via neutralisation.

EFFECT: obtaining a novel bleaching system.

11 cl, 2 ex

FIELD: chemistry.

SUBSTANCE: cleaning compositions contain organic catalysts having improved enzyme compatibility and having the following formulae: where each R1 independently denotes a branched alkyl group selected from a group comprising 2-butyloctyl, 2-pentylnonyl, 2-hexyldecyl, isodecyl, isotridecyl and isopentadecyl, or a linear alkyl group containing 11-18 carbon atoms. The composition also contains one or more auxiliary ingredients.

EFFECT: cleaning compositions with efficient bleaching at low water temperature.

15 cl, 16 ex

FIELD: chemistry.

SUBSTANCE: synthetic detergent with low environmental hazard contains sodium tripolyphosphate, anionic surfactant - sodium soap based on high fatty acids, carboxymethyl cellulose, sodium silicate, optical bleaching agent, fragrance component, sodium sulphate and water. The detergent also contains a nonionic surfactant - oxyethylated fatty alcohols, collagen dissolution products obtained by using leather production wastes (leather cuttings), sodium perborate and sodium carbonate. All components are taken in a defined ratio.

EFFECT: invention enables production of a synthetic detergent with low environmental hazard, good detergent action and low foaming capacity.

3 tbl, 3 ex

FIELD: chemistry.

SUBSTANCE: invention relates to pellets which contain diacyl peroxide particles inside a matrix, a method of preparing such pellets and detergents containing such pellets. The pellets contain a matrix made from at least one metal ion-cross-liked polysaccharide and diacyl peroxide particles put inside the said matrix. The pellets are obtained through drop-wise addition of an aqueous suspension in which diacyl peroxide particles are suspended and in which at least one metal ion-cross-linked polysaccharide is dissolved to a solution which contains polysaccharide cross-linking metal ions.

EFFECT: obtaining pellets in which diacyl peroxide is in a form which ensures its reliable production and storage.

11 cl, 1 ex

FIELD: chemistry.

SUBSTANCE: invention relates to aqueous liquid compositions for bleaching, cleaning and disinfecting surfaces. The invention describes an aqueous liquid bleaching composition which contains hypochlorite, a quaternary ammonium salt of formula: R1R2R3R4N+X-, where R1 - C10-C20 alkyl; R2, R3 and R4 - C1-C3 alkyl; X is an inorganic anion, and a viscousifying system which contains an amine oxide as a surfactant and a fatty acid. Described also is a method of imparting prolonged antibacterial activity on a solid surface using the said composition and a container for preparing the said composition.

EFFECT: obtaining a composition which retains its activity and stability during storage for 4 weeks.

16 cl, 2 tbl, 4 ex

FIELD: chemistry.

SUBSTANCE: invention relates to a method of cleaning at least part of a surface and/or fabric, involving: optional steps for washing and/or rinsing the surface and/or fabric; bringing the surface and/or fabric into contact with a washing solution containing a perhydrolase enzyme and a substrate for said enzyme, where the initial pH of the washing solution is alkaline and the amount of the perhydrolase enzyme and substrate is sufficient to lower the pH of the washing solution to 6.5 or lower; and optional washing and/or rinsing the surface and/or fabric, where said contact takes place during the washing cycle, and where lowering of the pH of the washing solution improves efficiency of the component of the washing solution.

EFFECT: improved method for bleaching textile.

7 cl, 5 ex, 5 dwg

FIELD: chemistry.

SUBSTANCE: composition contains more than 5% anionic surfactant, less than 25% nonionic surfactant, a light-sensitive ingredient and an inorganic mother-of-pearl agent. The light-sensitive ingredient is selected from a group comprising enzymes, dyes, vitamins, aromatising agents and mixtures thereof.

EFFECT: inorganic mother-of-pearl agent improves stability of light-sensitive ingredients in the detergent composition.

20 cl, 14 ex

FIELD: chemistry.

SUBSTANCE: invention discloses a composition which contains a fabric toning agent and a bacterial alkaline enzyme which exhibits endo-beta-1,4-glucanase activity (E.C.3.2.1.4). The fabric toning agent meets Testing method 1 requirements given in the description. Described also is a method of cleaning and/or processing a surface or fabric using the said composition.

EFFECT: composition ensures better removal of yellow stains and improved deposit of the agent with improvement of the outer appearance of the fabric.

18 cl, 16 ex

FIELD: chemistry.

SUBSTANCE: cleaning compositions contain organic catalysts having improved enzyme compatibility and having the following formulae: where each R1 independently denotes a branched alkyl group selected from a group comprising 2-butyloctyl, 2-pentylnonyl, 2-hexyldecyl, isodecyl, isotridecyl and isopentadecyl, or a linear alkyl group containing 11-18 carbon atoms. The composition also contains one or more auxiliary ingredients.

EFFECT: cleaning compositions with efficient bleaching at low water temperature.

15 cl, 16 ex

FIELD: chemistry.

SUBSTANCE: present invention relates to a composition which contains lipase, and a bleaching catalyst which can accept an oxygen atom from a peroxy acid and transfer the oxygen atom to an oxidisable substrate.

EFFECT: less bad smell which arises when using lipase enzymes in detergents.

13 cl, 4 tbl, 3 ex

FIELD: chemistry.

SUBSTANCE: detergent composition contains a polymer combined with a surfactant or an inorganic detergent component and auxiliary ingredients. The fat purification efficiency index of the detergent composition is at least equal to 10. The polymer is a random grafted copolymer which has a hydrophilic skeleton and hydrophobic side chains, obtained by grafting (a) polyethylene oxide; (b) vinyl ester of acetic and/or propionic acid; and/or C1-4alkyl ester of acrylic or methacrylic acid; and (c) modifying monomers. The detergent compositions preferably additionally contain lipase enzyme.

EFFECT: improved removal of fat contaminants and stains with reduced amounts of conventional surfactants or inorganic detergent components.

22 cl, 6 ex

FIELD: chemistry.

SUBSTANCE: compositions contain certain versions of lipase and a photo-bleaching agent based on xanthene dye, a photoinitiator and their mixtures.

EFFECT: more efficient cleaning, leading to minimal undesirable bad smells.

11 cl, 3 tbl, 16 ex

FIELD: chemistry.

SUBSTANCE: present invention relates to compositions which contain certain versions of lipase and a toned agent for fabric, and includes use of such compositions for cleaning and/or treating surface areas or fabric.

EFFECT: improved deposition of temporary colourant, reduced activity of the enzyme which leads to bad smell, and improved perception of cleanness.

17 cl, 3 tbl, 16 ex

FIELD: chemistry.

SUBSTANCE: detergent contains the components as follows, wt %: anionic surface active agent (SAG) - alkylbenzol sodium sulphonate 10-16; nonionic SAG - oxyethylated fatty alcohols or oxyethylated alkylphenol 2-5; sodium tripolyphosphate 15-25; organophosphonate compound - sodium salt 1-hydroxyethylidene of phosphonic acid or sodium diethylentriaminopentaxys-(methylene phosphonate) 0.2-0.6; polycarboxylate 0.5-1.5; carboxymethyl cellulose 0.3-0.6; modified polyalkylene glycol 0.2-0.6; optical bleaching agent 0.05-0.3; soda ash 3-6; liquid glass 3.5-6.0; enzyme 0.4-0.7; defoaming agent 0.05-1.5; aromatiser 0.15-0.3; sodium sulphate and water to 100.

EFFECT: higher efficiency of bleaching detergent, all type decontamination with additional softening of fabric and without irritation of hand skin during manual washing, lower temperature modes of washing, reduced damage effect on metal parts of washing machines.

10 cl, 2 tbl, 5 ex

FIELD: chemistry.

SUBSTANCE: invention is meant for soaking, washing and bleaching all types of textile articles, except articles from natural silk and wool, in any kind of machine, and also hand wash. Substance contains in % mass: an anionic surfactant 7-15, nonionic surfactant oxyethylated fatty alcohol 2-5, sodium tripolyphosphate 15-25, sodium ethylene-diaminotetraacetate (versene) 0.3-1, polycarboxylate, containing monomers of acrylic acid 0.2-0.6, carboxymethyl cellulose (CMC) (in terms of 100% of the share of the main substance) 0.4-0.6, sodium silicate (in terms of SiO2) - 2-5, sodium perborate or sodium percarbonate (in terms of activated oxygen) 1.5-4, tetraacetylethylenediamine (TAED) 0.5-3, enzymes 0.4-1, caustic ash 5-15, mixture of sodium carbonate and amorphous sodium silicate 2-5, optical brightener 0.05-0.4, perfume 0.1-0.3, sodium sulphate and water till 100.

EFFECT: increase in the effectiveness of the detergent with a whitening effect, removal of all kinds of dirt while adding extra softness without an irritating effect on the skin during hand washing, reducing the temperature regimes of washing, reduction in the harmful effect to the metallic part of the washing machine.

9 cl, 2 tbl

FIELD: gene and protein engineering, in particular substances for detergents and cleaning compositions.

SUBSTANCE: invention relates to mutant forms of subtilisin Bacillus, obtained by certain combination of replacement in origin native enzyme amino acid sequence. Such mutant forms are differ from precursor subtilisin by increased cleaning effect. According to present invention all subtilisin variants are characterized either by residue substitution in position accepted to 232 position of Bacillus amyloliquifaciens amino acid sequence with valine, or by residue substitution in position accepted to 212 position of said subtilisin natural form with proline. To produce disclosed new form of enzyme respective mutant DNA sequences have been expressed in host cells, preferably in strain Bacillus cells with decreased level of proteases activity. Subtilisin variants of present invention are useful as components of any cleaning composition allowing for protease including.

EFFECT: new materials for detergents and cleaning compositions of improved effect.

6 cl, 5 dwg, 6 tbl, 3 ex

Up!