DE10358535A1 - Hybrid enzymes with cationic binding domain - Google Patents

Abstract

The present invention relates to hybrid enzymes comprising a hydrolytically active moiety and a cationic binding moiety, as well as formulations containing these hybrid enzymes.

Description

The The present invention relates to hybrid enzymes comprising a hydrolytic active unit and a cationic binding unit and preparations, containing these hybrid enzymes.

When Biofilm refers to the growth of surfaces by microbes and before especially the gelatinous ones they collectively give and envelop Protective layer, also called EPS matrix, where EPS stands for "extracellular polymers Substances "stands. This protective layer is made up of, inter alia, together Proteins and polysaccharides, in the most diverse dirt particles such as dead cells, skin particles and lime stored are. This mucus is also the reason that under real living bacteria, as well as other biofilms producing Microorganisms, much more resistant to - even biocidal - cleaning and disinfectants are as laboratory cultures, usually from individual free-floating bacteria, so-called planktonic cells, consist. Lead biofilms in technical systems to massive impairments and can in the medicine persistent infections cause the opposite usual Antibiotics are largely resistant. In the household are the through Biofilms induced impairments both more hygienic and more aesthetic Nature.

The fight biofilm biocides is only conditionally possible, since the majority polysaccharide mucus matrix precedes the cells Influence of biocides protects (so-called pseudoresistance). At all to achieve an effect would have the biocides therefore much higher be used metered, as it is for a comparable amount Unassociated present bacteria would be needed. The use of biocides in the household, however, is under ecological Unfavorable so the aspirations are more likely to go with detergent to provide only low concentrations or completely without biocides. This let himself go but so far not with the desire for a complete removal bacterial biofilms - and thus optimal hygiene - in Bring harmony.

The European patent EP 946 207 B1 (corresponding to WO 98/26807) teaches the enzymatic treatment of biofilms with combinations of hydrolases which serve to detach and remove the biofilms from the surface and oxidoreductases which kill the contained bacteria. These systems clean and disinfect hard surfaces made of a wide variety of materials in various industrial sectors, such as cooling towers, dairies or chemical plants, as well as soft surfaces such as skin, hair or textiles.

EP 388 115 deals with the destruction and removal of slime on industrial, water-washed surfaces. For this purpose, glucanases, cellulases, amylases and / or proteases are used which specifically decompose the polysaccharides of the mucus matrix; Subsequently, the now accessible microorganisms should be exposed to the action of biocides.

In WO 01/09295 are enzymes having a β- (1,6) -Endoglucanaseaktivität and corresponding DNA sequences claimed, further an enzyme preparation for degradation β-1,6-glucanhaltiger Materials and their use. The β- (1,6) -endoglucanases are also used for removal of biofilms of different surfaces. Mainly deals However, this Spanish writing with the degradation of β-1,6-glucanreichen Cell wall of certain fungi.

Also WO-PS 96/36569 describes an enzymatic method for Removal of biofilms. This will be a mannanase, if necessary used in combination with at least one other enzyme, in order to aquatic in industrial Systems to prevent slime formation and existing mucus to remove. According to WO 96/17632 This task can also be achieved with combinations of short-chain glycol components with at least one enzyme from the group of polysaccharidases, proteases, Dissolved lipases and glycoproteases become.

The International Application WO 01/53010 also teaches an enzymatic Process to Removal of biofilms from various surfaces washed with water or aqueous solutions industrial systems as well as in the medical field, where the biofilms both above the water surface, ie at solid / air interfaces, as also under water, at solid / liquid interfaces. at the enzymes used are carbohydrases or proteases, Combinations of these two enzymes are also possible. Possibly can the cleaning solution in addition to the enzymes mentioned further active substances such as corrosion inhibitors, Surfactants, biocides, etc. included.

As recently (Whitchurch et al. (2000) Science 295, 1487ff), bacterial biofilms also consist of nucleic acids and are particular in early Stages also by nucleic acid-degrading Enzymes, e.g. hydrolyzed by DNAses.

The enzymatic hydrolysis of the biofilm by a hydrolase-containing Cleaner runs However, in many cases very inefficient, because the exposure time through the Cleaning process is very short, the enzymes washed out quickly and thus not sufficiently long in contact with the polymers stand. As a result, only a small or incomplete Hydrolyseleistung achieved.

task The present invention was therefore a method for effective To provide removal of the biofilm, especially one such more effective than previously described in the prior art Method.

It were here considerations hired hydrolytic enzymes to be an efficient means of Could be removal of the biofilm, if their residence time increases could be.

task The present invention was therefore particularly a method to disposal to adjust, with which the residence time of for removal of the biofilm suitable hydrolases on the surface is increased.

task Further, the present invention has been more particularly a detergent that can be delivered without the use of biocides, Biofilms on hard surfaces dissolve in the household and to remove the film-forming microorganisms.

The Main task is solved enzymes that are able to stay longer on the surface and therefore capable of more effective biofilm degradation.

Is solved this task in particular by such enzymes, the one Have modification in the form of a cationic binding domain.

by virtue of The negative charge of the biofilm can be modified in this way Enzymes longer on Biofilm or on surfaces, which have a biofilm, adhere and thus more effectively the biofilm dismantle.

These Hybrid enzymes are capable of doing so, the bacterial mucus matrix in biofilms on toilets and other surfaces to destroy, so that the bacteria are released from their dressing and into the planktonic Be transferred form, causing a simple flushing sufficient to remove the microbes, without the addition of the Need to use biocides would. It is therefore an environmentally friendly method for cleaning and disinfecting especially moist, wasserumspülter surfaces accessible become

Hybrid proteins, the binding domains are already described in the prior art.

So WO 95/31556 describes hybrid proteins from glucan binding domains the antimicrobials are bound for use as oral care products. WO 98/18437 describes mutan binding domains to which antimicrobial Substances are also bound for use as oral care products. In WO 98/16190 are hybrid proteins from starch binding domains with described antimicrobial substances, which are also used as oral care products Find use. In WO 99/33957 modified enzymes are described, the one polyanionic domain which are also used in oral care products.

WHERE 99/18999 also describes compositions used in oral care products Use, wherein the composition comprises an enzyme as well an anchor molecule, which is bound to the enzyme and is capable of binding to a substrate to tie up that close located in a bacterial colony. In the embodiments are as anchor molecules short-chain basic and acidic molecules.

WO 01/93875 describes compositions for treating biofilm structures, the composition comprising an enzyme capable of degrading biofilm structures and an anchor capable of binding to the cellular colony or to other bioadhesive molecules. When Examples of the anchor molecule include the binding domain of elastase, domains that bind to carbohydrates and polysaccharides, lectins, selectins, the binding domain of heparin, the binding domain of fibronectin, and the CD44 protein.

US 2002022005 describes a composition for degrading biofilm structures associated with cystic fibrosis, the composition comprising an enzyme capable of degrading biofilms and comprising an anchor capable of binding to or on the surface Close to the biofilm structure. As anchor molecules, for example, concanavalin, lectins, elastase, heparin and amylose-binding domains are mentioned.

WHERE 00/47174 and WO 00/47175 describe oral care products containing hybrid proteins containing an enzyme and an anchor molecule. With the enzymes these are those which can degrade polysaccharides the anchor molecule is it a molecule, that is capable of doing so "in the Near one bacterial colony " tie. As examples of the anchor molecule are glutathione-S-transferase, Glutathione, avidin, streptavidin, biotin and antibodies.

Hybrid proteins according to the invention as well as the use according to the invention however, the hybrid proteins have been in the art so far not described.

One The first subject of the present invention are therefore hybrid proteins, comprising a hydrolytically active unit and a cationic one Bond unit.

at the hydrolytically active unit of the hybrid enzyme according to the invention it is preferably a hydrolytic enzyme, especially from the group of polysaccharidases, in particular β-glucanases, Cellulases, xylanases, amylases, dextranases, glucosidases, galactosidases, Pectinases, chitinases or alginate lyases or lysozyme, and / or from the group of proteases and peptidases, in particular subtilisin, Thermolysin, pepsin, carboxypeptidase or acid protease, and / or from the group of nucleases, in particular DNAses or RNAses and / or it is a functional fragment thereof.

at The cationic binding moiety may be a natural one occurring or to an artificial one prepared binding domain act. Preferably, the binding domain is a protein or a polypeptide such that the hybrid enzyme is a Fusion protein acts. Alternatively, however, other units may be used used as cationic binding unit or cationic binding domain which have a positive charge, for example organic molecules carry the amino groups. In the case of proteins or polypeptides is the binding domain preferably not more than 150 amino acids in length, more preferably Does she have a length? between 10 and 140 amino acids. The average number of charges is preferably between 0.01 and 0.8, more preferably between 0.1 and 0.4 positive Charges per amino acid residue at neutral pH.

• a) natürlicherweise vorkommende oder künstlich synthetisierte Mutanten der genannten Polypeptide, insbesondere solche, die bis zu zehn, bevorzugt bis zu fünf, vor allem genau ein, zwei oder drei Punktmutationen in Bezug auf die genannten Polypeptide besitzen,
• b) Polypeptide, die eine Sequenzhomologie oder -identität von mindestens 50 %, bevorzugt mindestens 60 % oder 70 %, ganz besonders bevorzugt mindestens 80 oder 90 %, insbesondere mindestens 95 % oder 98 % in Bezug auf die genannten Polypeptide besitzen,
• c) Polypeptide bestehend aus mindestens 10 oder 15, besonders bevorzugt mindestens 20 oder 30, aufeinanderfolgenden Aminosäuren der genannten Polypeptide oder eines Polypeptids gemäss (a) oder (b),
• d) Polypeptide mit Insertionen und/oder Deletionen von bis zu 10, 5 oder 3 Aminosäuren in Bezug auf die genannten Polypeptide oder in Bezug auf ein Polypeptid gemäss (a), (b) oder (c).

In a preferred embodiment, the binding domain comprises a binding domain of a naturally occurring surface protein, in particular a so-called cell envelope protein, a cell membrane-associated protein and / or a so-called surface layer protein (S-layer protein) and / or a fragment and / or derivative thereof. For example, this may be the binding domain of an sbs protein, in particular a sbs protein from Geobacillus, in particular from Geobacillus stearothermophilus, or a fragment and / or derivative thereof. The binding domain here may in particular comprise a polypeptide having an amino acid sequence according to SEQ ID NO: 19 or according to SEQ ID NO: 20 or derivatives of said polypeptides. Derivatives are to be understood as meaning in particular:

• a) naturally occurring or artificially synthesized mutants of said polypeptides, in particular those having up to ten, preferably up to five, in particular exactly one, two or three point mutations with respect to said polypeptides,
• b) polypeptides having a sequence homology or identity of at least 50%, preferably at least 60% or 70%, most preferably at least 80 or 90%, in particular at least 95% or 98% with respect to the said polypeptides,
• c) polypeptides consisting of at least 10 or 15, more preferably at least 20 or 30, consecutive amino acids of said polypeptides or of a polypeptide according to (a) or (b),
• d) polypeptides having insertions and / or deletions of up to 10, 5 or 3 amino acids with respect to said polypeptides or with respect to a polypeptide according to (a), (b) or (c).

Further surface layer proteins which can be used according to the invention are, for example, Sára and Sleytr (2000, Journal of Bacteriology 182 (4), 859-868).

• a) natürlicherweise vorkommende oder künstlich synthetisierte Mutanten eines Polypeptids mit einer Aminosäuresequenz gemäss SEQ ID NO: 21, insbesondere solche, die bis zu zehn, bevorzugt bis zu fünf, vor allem genau ein, zwei oder drei Punktmutationen in Bezug auf ein Polypeptid mit einer Aminosäuresequenz gemäss SEQ ID NO: 21 besitzen,
• b) Polypeptide, die eine Sequenzhomologie oder -identität von mindestens 50 %, bevorzugt mindestens 60 % oder 70 %, ganz besonders bevorzugt mindestens 80 oder 90 %, insbesondere mindestens 95 % oder 98 % in Bezug auf ein Polypeptid mit einer Aminosäuresequenz gemäss SEQ ID NO: 21 besitzen,
• c) Polypeptide bestehend aus mindestens 10 oder 15, besonders bevorzugt mindestens 20 oder 30, aufeinanderfolgenden Aminosäuren eines Polypeptids mit einer Aminosäuresequenz gemäß SEQ ID NO: 21 oder eines Polypeptids gemäss (a) oder (b),
• d) Polypeptide mit Insertionen und/oder Deletionen von bis zu 10, 5 oder 3 Aminosäuren in Bezug auf ein Polypeptid mit einer Aminosäuresequenz gemäß SEQ ID NO: 21 oder in Bezug auf ein Polypeptid gemäss (a), (b) oder (c).

In a further preferred embodiment, the cationic binding domain is an artificially produced polypeptide, especially one having a length between 25 and 40 amino acids, which preferably carries between 0.3 and 0.4 positive charges at neutral pH, wherein the positively charged groups are preferably lysine residues. The cationic binding domain here preferably comprises a polypeptide having an amino acid sequence according to SEQ ID NO. 21 or a derivative thereof, wherein a derivative is to be understood in particular

• a) naturally occurring or artificially synthesized mutants of a polypeptide having an amino acid sequence according to SEQ ID NO: 21, in particular those having up to ten, preferably up to five, especially one, two or three point mutations with respect to a polypeptide having an amino acid sequence according to SEQ ID NO: 21,
• b) polypeptides having a sequence homology or identity of at least 50%, preferably at least 60% or 70%, most preferably at least 80 or 90%, in particular at least 95% or 98% with respect to a polypeptide having an amino acid sequence according to SEQ ID NO: 21,
• c) polypeptides consisting of at least 10 or 15, more preferably at least 20 or 30, consecutive amino acids of a polypeptide having an amino acid sequence according to SEQ ID NO: 21 or a polypeptide according to (a) or (b),
• d) polypeptides having insertions and / or deletions of up to 10, 5 or 3 amino acids with respect to a polypeptide having an amino acid sequence according to SEQ ID NO: 21 or with respect to a polypeptide according to (a), (b) or (c) ,

object The present invention furthermore relates to nucleic acids which are suitable for one of the abovementioned polypeptides encode.

object The present invention furthermore relates to aqueous preparations (enzyme preparations), the hybrid proteins of the invention in dissolved Form included.

object The present invention further relates to the use of the hybrid enzyme of the invention and / or enzyme preparations containing hybrid enzymes, in particular in sterilization, disinfection, washing and cleaning agents, especially in cleaners to remove biofilms from hard surfaces. Of the Use of funds may be particularly in the household or in the field Pharmaceuticals, Food, Brewery, Medical Engineering, Color, Wood, textile, cosmetic, leather, tobacco, fur, rope, paper, pulp, Plastic, fuel, oil, Rubber or machinery industry or in molecular operations. The hybrid enzymes according to the invention can especially for the removal of biofilms from household and / or sanitary items as well in pulp and paper mills, in circulating cooling towers as well in other systems, the flowing one and / or circulating water, be used. In addition to the removal of biofilms are the hybrid enzymes of the invention for example, also suitable for use in detergents for cleaning of textiles, in particular for stain removal. By the positive Binddomain is facilitates adhesion to negatively charged textile surfaces.

object sterilizing, disinfecting, Detergents and cleaning agents, the hybrid enzymes of the invention and / or hybrid enzymes containing enzyme preparations. The remedy may be basically a means for act any kind of surface. So it may be at the surface a biotic or abiotic, artificially synthesized or natural, soft or hard surface act. At the surface it may be about a textile, ceramic, metal and / or plastic surface. The article can be, for example, laundry, sanitary facilities, flooring, Shoes, leather, articles made of rubber, hulls, prostheses, Teeth, Dentures or catheters act.

at The cleaning agent may in particular be a household cleaner or a cleaner for industrial installations, in particular those mentioned above. In a preferred embodiment it is the cleaner to such a for cleaning hard surfaces, such as for example, of soils, Tiles, tiles, plastics and other hard surfaces in the Household, in industrial plants, in public sanitary facilities, in swimming pools, Saunas, sports facilities or in doctor or massage practices.

Medically relevant biofilms which can be removed with hybrid enzymes according to the invention are, for example, biofilms due to chronic lung infections, which can occur, for example, in cystic fibrosis patients. Furthermore, it may be plaque or biofilms, the contact lenses, Imp lantates or medical devices, instruments or apparatus such as catheters or endoscopes.

object The present invention therefore also relates to the use of the hybrid enzymes according to the invention and / or hybrid enzyme-containing enzyme preparations in cosmetic and / or pharmaceutical products as well as cosmetic and / or pharmaceutical Products containing the hybrid enzymes of the invention and / or enzyme preparations and further the use the hybrid enzymes according to the invention and / or enzyme preparations for the production of cosmetic and / or pharmaceutical products. For example, the product may be to get a product for removing plaque from tooth surfaces or a product for the removal of biofilms caused by chronic Lung infections caused or associated with these, such as e.g. in case of cystic fibrosis, act. Subject of the present Invention are therefore especially oral, dental or denture care, the the hybrid enzymes according to the invention contain.

The Sterilization, disinfection, washing and cleaning agents can further Contain components as they are known in the art, in particular one or more components selected from the group consisting from surfactants, builders, acids, alkaline substances, hydrotropes, solvents, thickeners, Dyes, perfumes, corrosion inhibitors and skin protection agents. If the detergent sprayed If appropriate, a propellant may also be present. The cleaning agent is preferably a liquid aqueous agent, it may be but also, for example, a gel, a paste or a powder act.

One Another object of the invention is a product containing an enzyme preparation according to the invention or a detergent according to the invention and a spray dispenser.

Prefers is the spray dispenser a manually activated spray dispenser, especially selected from the group comprising aerosol spray dispensers (pressurized gas containers; et al referred to as spray can), even pressure-building spray dispensers, Pumpsprühspender and trigger spray dispensers, in particular pump spray dispenser and trigger spray dispensers with a container made of transparent polyethylene or polyethylene terephthalate. spray dispenser become more detailed in WO 96/04940 (Procter & Gamble) and the spokesmen in it cite US patents, to which reference is made in this regard and its contents are hereby incorporated into this application is described. Triggersprühspender and pump sprayer own opposite Compressed gas containers the advantage that no Propellant must be used.

In a further preferred embodiment however, the agent containing the enzyme preparation will not be considered as Aerosol atomized, here possibly small amounts of the enzyme preparation in the respiratory tract can be advised and possibly trigger allergic reactions there could. By suitable, particle-permeable Essays, Nozzles etc. (so-called "nozzle valves") on the spray dispenser For example, in enzyme-immobilized form, the enzyme may be the agent enclosed be dosed and as cleaning foam. In the production of these compact foams no respirable occur Particles, so that the Danger of inhalation of allergens is essentially banned.

Yet Another subject of the invention is a method for hydrolysis biofilms on hard surfaces, in which the vitality the microbial cells are not damaged. For this purpose, the inventive enzyme preparation or the cleaning agent according to the invention, optionally using a product according to the invention, on the biofilm-coated area applied and optionally after a contact time of 1 to 30 minutes, with stronger Infest also for a few hours or over Night, rinsed with clean water. The preparation or the agent may also be washed before rinsing a cloth, a sponge, a brush or some other Cleaning suitable utensil on the biofilm infested surface blurs or triturated, which is e.g. in the presence of abrasives Improved cleaning performance in the cleaning agent. Finally will the surface dried with a dry cloth. In case of very strong infestation of surface with biofilm, the cleaning process can then be repeated.

Substances which also serve as ingredients of cosmetic products are referred to below, where appropriate, according to the International Nomenclature Cosmetic Ingredient (INCI) nomenclature. Chemical compounds carry an INCI name in English, plant ingredients are listed exclusively in Linnaeus in Latin, so-called trivial names such as "water", "honey" or "sea salt" are also given in Latin. The INCI names can be found in the International Cosmetic Ingredient Dictionary and Handbook - Seventh Edition (1997), published by The Cosmetic, Toiletry, and Fragrance Association (CTFA), 1101 17th Street, NW, Suite 300, Washington, DC, 20036, USA, and more than 9,000 INCI names, as well as references to more than 37,000 trade names and technical names, including related distributors, from over 31 countries contains. The International Cosmetic Ingredient Dictionary and Handbook assigns to the ingredients one or more chemical classes, such as polymeric ethers, and one or more functions, such as surfactants-cleansing agents, which are further explained and discussed below possibly also referred to.

The Specification CAS means that it in the following sequence of numbers to a name of the Chemical Abstracts Service is.

enzyme preparation

The inventive enzyme preparation is a system containing one or more of the hybrid enzymes of the invention. If necessary, you can also other unmodified enzymes, in particular selected from Polysaccharidases, proteases and nucleases in the enzyme preparation be included. The following explanations refer, as far as is not explicitly distinguished, both on the free enzymes as also on the enzymes that are part of the hybrid enzymes.

When Polysaccharidases are enzymes which are hydrolytic Catalyze cleavage of polysaccharides. This is what the cellulase does For example, degraded cellulose, while the glucosidase from the Oligo- or polysaccharide molecule splits off individual glucose units from the end. The extracellular polysaccharide matrix of biofilm consists of homo- and heteropolysaccharides, which are all from glucose, fucose, mannose, galactose, fructose, pyruvate, mannuronic and glucuronic acid are constructed. Accordingly, it is the polysaccharides for example, levans, polymannans, dextrans, cellulose, amylopectin, Glycogen or alginate.

in the All of the purposes of this invention Polysaccharidases used for the hydrolysis of biofilms. Prefers contains the enzyme preparation, however, at least one enzyme from the group β-glucanase, Cellulase, xylanase, amylase, dextranase, glucosidase, galactosidase, Pectinase, chitinase, lysozyme, hemicellulase and alginate lyase. These enzymes can each include several family members. So you know, for example both an α-1,4-glucosidase, the sooner also referred to as maltase, as well as a β-1,4-glucosidase. For the purposes of this invention, all these subgroups are included. It can too already commercially available Mixtures of polysaccharidases can be used. So is suitable For example, Viscozyme, a liquid enzyme preparation Aspergillus sp., Composed of different polysaccharidases is, i.a. from arabanase, cellulase, β-glucanase, hemicellulase and Xylanase, for use in the context of this invention.

proteases are enzymes that promote the hydrolytic cleavage of the peptide bond catalyze proteins and peptides. They can therefore also be broken down used by biofilms, as these in addition to polysaccharides as well proteins secreted by the film-forming organisms can contain. Proteases can systematically, on the one hand, according to the pH optimum of its effect in acid, neutral and alkaline proteases are divided, on the other hand but classified according to the catalytically active group in the active center. A distinction is made between serine, cysteine, aspartate and metalloproteases.

All known proteases can be used in the context of this invention for the degradation of biofilms, however, preferably at least one protease is selected from the group subtilisin, thermolysin, pepsin, carboxypeptidase, acid Protease used.

at The nucleases are nucleic acid degrading enzymes. Depending on Hydrolysis site distinguishes endonucleases that are within the nucleic acid strand cleavage (e.g., pancreatic DNAse I) and exonucleases, the nucleotides release from the end (e.g., exonuclease III from E. coli). there can DNA single strands (S1 nuclease from Aspergillus oryzae) or double strands (exonuclease III) serve as a substrate and quite nonspecific to shorter strands or individual nucleotides are degraded. Also mixed specificities are possible and occur in dependence z. Of enzyme concentration and presence of certain ions. Similarly, the cleavage of RNA molecules is possible. Highly specific endonucleases require certain nucleotide sequences (typically 4-8 base pairs) and in part also as restriction enzymes or restriction endonucleases designated. Since biofilms can also consist of nucleic acids are also all Nucleases for use according to the invention suitable in biofilm degrading agents. However, preferably unspecific DNAses or RNAses used.

The In the context of this invention usable non-modified enzymes can by usual biochemical methods obtained from the corresponding organisms become. But they are also commercially available, for example from the companies Novozymes, Genencor International, Sigma, AB Enzymes or ASA enzymes. It can the enzyme preparations also of technical quality and consist of several different enzyme activities. The presence from partially not closer characterized ancillary activities may be due to the hydrolysis of complex biofilm polymers also have a favorable influence.

Should Proteases with other enzymes, for example polysaccharidases, be combined, it should be noted, regardless of whether they are hybrid enzymes according to the invention or non-modified enzymes that these are in liquid agents by components contained in the cleaner such as surfactants or organic acids or by the rather nonspecific acting proteases could be attacked which reduces the effectiveness of the agents over non-protein biofilm constituents would reduce. To the activity and stability such cleaning agents over a longer one To ensure a period of time Therefore, it makes sense to use sensitive enzymes of denaturing components and in particular also proteases separated from other enzymes hold so that this Components only immediately before or during the cleaning process meet each other.

These Separation of the different enzymes can be done in different ways. A possibility is to introduce the enzymes into microcapsules that are only immediate before or during the cleaning process, for example by mechanical action during the cleaning process, by sudden change the osmotic conditions e.g. when diluting with water or by cutting the capsules when dosing from the storage bottle, opened and release their content. However, it should be noted that the capsule wall in turn is not attacked by the enzymes may be. Capsules based on proteins or Polysaccharides are therefore excluded. Alternatively, the Enzymes are each immobilized on a solid support, the is suspended in the detergent and in the application of the agent additionally can serve as abrasive. The immobilization takes place on the Specialist way known; as carrier materials can in addition to inorganic substances, for example silicates or porous glass, also polymers are used.

A another conceivable possibility could also be incorporating into a multiphasic means, so that the Protease predominantly located in one phase and the remaining enzymes predominantly in another phase. Such a remedy would shaken just before use and applying the resulting emulsion to the surface to be cleaned; the remaining in the reservoir bottle would then a experienced rapid phase separation, so that the enzymes only for short Time and possibly come in contact at the phase interface. The case would be similar stored, if an enzyme would be incorporated into a phase, the in the form of droplets in the second, the other enzyme-containing phase stably dispersed would; in this connection that would be Medium not shaken, but dosed directly from the storage bottle, possibly also sprayed. However, such a multi-phase means would be a technically complex and difficult-to-implement solution. In addition, would be consumer acceptance is expected to be low.

meaningful it is therefore, the two enzymes or enzyme groups in spatially separated solutions store. The storage in two completely separate bottles is the less preferred solution, since this is for the consumer a greater effort means. Therefore, the provision in one is particularly preferred Two-chamber bottle, from which then dosed simultaneously both cleaning solutions can be. Such two-chamber bottles are already known from the prior art known. Besides the possibility provides separate storage of incompatible enzymes a two- or multi-chamber bottle further advantages. So it is possible in a chamber, inter alia, an enzyme in an optimal pH solution with respect to storage stability to store and in another chamber a solution store, whose pH leads to that at Mixing this with the enzyme storage solution creates a cleaning solution, their pH causes an optimum of enzyme activity. For example many polysaccharidases are particularly stable at neutral pH, unfold their highest activity but at pH 4 to 5.

cleaning supplies

The For the purposes of this invention, the enzyme preparation can also be used in a cleaning agent for hard surfaces be used. The cleaning agent may be the enzyme preparation in amounts of 0.1 to 10.0 wt .-%, preferably 0.5 to 3% by weight. The cleaning agent here is preferably biocide-free, can but may also contain small quantities of biocide.

Next the enzyme preparation can be a liquid, gel-like or pasty aqueous cleaning agent for hard surfaces According to the invention also customary detergent ingredients contain. These ingredients include, for example, surfactants, but also builders, acids, Alkalis, hydrotropes, solvents, thickeners, Abrasives and other auxiliaries and additives such as dyes, Perfumes, corrosion inhibitors or skin care products.

The optionally used surfactants are preferably selected from the group anionic surfactants, nonionic surfactants and / or amphoteric Surfactants. In this case, preference is given to anionic and / or nonionic Surfactants used. If used, come anionic Surfactants preferably in amounts of 0.1 to 15 wt .-%, nonionic Surfactants preferably in amounts of 0.1 to 10 wt .-% and amphoteric Surfactants are preferably used in amounts of from 0.1 to 4% by weight, in each case based on the total composition. In a less preferred embodiment can In addition, cationic surfactants in amounts of up to 2 wt .-% used, based on the total composition. As well can also cationic surfactants are used. In a preferred embodiment is the detergent due to the cationic surfactants outgoing biocidal action, however, free of these.

To the inventively usable include anionic surfactants aliphatic sulfates such as fatty alcohol sulfates, fatty alcohol ether sulfates, Dialkyl ether sulfates, monoglyceride sulfates and aliphatic sulfonates such as alkanesulfonates, α-olefinsulfonates, Ether sulfonates, n-alkyl ether sulfonates, sulfonated fatty acids, ester sulfonates and lignosulfonates. Also in the context of the present invention usable are alkylbenzenesulfonates, fatty acid salts (soaps), fatty acid cyanamides, Sulfosuccinates (sulfosuccinic acid mono- and dialkyl esters), sulfosuccinamates, sulfosuccinamides, carboxylic acid amide ether sulfates, Alkyl polyglycol ether carboxylates, fatty acid isethionates, acylaminoalkanesulfonates (Taurides, N-acyl taurides), fatty acid sarcosinates, ether carboxylic acids and Alkyl (ether) phosphates and α-sulfo fatty acid salts, Acylglutamates, monoglyceride disulfates and alkyl ethers of glycerol disulfate, and finally their mixtures.

in the The present invention relates to fatty acids or fatty alcohols or their derivatives - insofar not stated otherwise - representative for branched or unbranched carboxylic acids or alcohols or their derivatives having preferably 6 to 22 carbon atoms, in particular 8 to 20 carbon atoms, particularly preferably 10 to 18 carbon atoms, most preferred 12 to 16 carbon atoms, for example 12 to 14 carbon atoms. The fatty acids / alcohols or their derivatives with even number of carbon atoms are in particular because of their vegetable base than on renewable resources based on ecological establish preferred, but without limiting the teaching of the invention to them. Especially are also available, for example, according to the ROELEN's oxo synthesis Oxo-alcohols or their derivatives having preferably 7 to 19 carbon atoms, in particular 9 to 19 carbon atoms, particularly preferably 9 to 17 carbon atoms, most preferred 11 to 15 carbon atoms, for example 9 to 11, 12 to 15 or 13 to 15 carbon atoms, can be used accordingly.

They are in the form of their alkali metal and alkaline earth metal salts, in particular sodium, potassium and magnesium salts, as well as ammonium and mono-, di-, tri- or tetraalkylammonium salts and in the case of the sulfonates also in the form of their corresponding acid, for example dodecylbenzenesulfonic acid, used. Due to their production, the alkyl ether sulfates always contain residual contents of non-alkoxylated fatty alcohol sulfates, especially at low degrees of ethoxylation. Furthermore, the compositions according to the invention may also comprise soaps, ie alkali or ammonium salts of saturated or unsaturated C ₆ -C ₂₂ -fatty acids.

Prefers The anionic surfactants are selected from the group comprising Fatty alcohol sulfates in amounts of up to 5% by weight, alkylbenzenesulfonates in amounts of up to 7.5% by weight and soaps in amounts of up to 2 wt .-%, each based on the total composition, and mixtures the same.

Suitable nonionic surfactants are, for example, C ₈ -C ₁₈ -alkyl alcohol polyglycol ethers, alkyl polyglycosides and nitrogen-containing surfactants or mixtures thereof, in particular the first two. C ₈ -C ₁₈ alkyl alcohol polypropylene glycol / polyethylene glycol ethers may be represented by the formula R ¹ O- (CH ₂ CH (CH ₃ ) O) _p (CH ₂ CH ₂ O) _e -H in which R ^{1 is} a linear or branched, aliphatic alkyl and / or alkenyl radical having 8 to 18 carbon atoms, p is 0 or numbers from 1 to 3 and e is numbers from 1 to 20. They are obtained by addition of propylene oxide and / or ethylene oxide to alkyl alcohols, preferably to fatty alcohols. Furthermore, end-capped C ₈ -C ₁₈ alkyl alcohol polyglycol ethers can be used ie, alkyl alcohol polyalkylene glycol ethers according to the above formula in which the free OH group is etherified.

Inventive cleaner can Alkyl alcohol polyglycol ether in amounts of 0.1 to 4 wt .-%, based on the total composition.

Preferred nonionic surfactants are also alkyl polyglycosides (APG) of the formula R ² O [G] _x , in which R ^{2 is} a linear or branched, saturated or unsaturated alkyl radical having 8 to 22 carbon atoms, [G] is a glycosidically linked sugar radical and x is a number from 1 to 10. The index number x indicates the degree of oligomerization (DP degree), ie the distribution of monoglycerides and oligoglycosides. Whereas x must always be an integer in a given compound, and above all the values x = 1 to 6, the value x for a given alkyl glycoside can be an analytically determined arithmetic variable, which usually represents a fractional number. Preferably, alkyl glycosides having a mean degree of oligomerization x of 1.1 to 3.0 are used. From an application point of view, those alkyl glycosides whose degree of oligomerization is less than 1.7 and in particular between 1.2 and 1.6 are preferred. The glycosidic sugar used is preferably xylose, but especially glucose. The alkyl or alkenyl radical R ² can be derived from primary alcohols having 8 to 18, preferably 8 to 14 carbon atoms. Typical examples are caproic alcohol, caprylic alcohol, capric alcohol and undecyl alcohol, and technical mixtures thereof, such as those obtained during the hydrogenation of technical fatty acid methyl esters or in the hydrogenation of aldehydes from ROELEN's oxosynthesis. Preferably, however, the alkyl or alkenyl radical R ² is derived from lauryl alcohol, myristyl alcohol, cetyl alcohol, palmoleyl alcohol, stearyl alcohol, isostearyl alcohol or oleyl alcohol. Also to be mentioned are elaidyl alcohol, petroselinyl alcohol, arachidyl alcohol, gadoleyl alcohol, behenyl alcohol, erucyl alcohol and technical mixtures thereof.

Alkylpolyglykoside can in cleaners according to the invention in amounts of 0.1 to 6 wt .-%, based on the total composition.

When other nonionic surfactants can containing nitrogen-containing surfactants, e.g. fatty acid polyhydroxy amides, For example, glucamides, and ethoxylates of alkylamines, vicinalen Diols and / or carboxylic acid amides, the alkyl groups having 10 to 22 carbon atoms, preferably 12 to 18 C atoms, own. The degree of ethoxylation of these compounds is usually between 1 and 20, preferably between 3 and 10. Preferred are ethanolamide derivatives of alkanoic acids with 8 to 22 C atoms, preferably 12 to 16 C atoms. To the particular include suitable compounds the lauric acid, myristic and palmitic acid monoethanolamides.

The amphoteric surfactants (amphoteric surfactants, zwitterionic surfactants) which can be used according to the invention include, inter alia, betaines, amine oxides, alkylamidoalkylamines, alkyl-substituted amino acids and acylated amino acids. Preferred amphoteric surfactants are, for example, betaines of the formula (R ³ ) (R ⁴ ) (R ⁵ ) N ⁺ CH ₂ COO ^- , in which R ^{3 is} an optionally interrupted by hetero atoms or heteroatom groups alkyl radical having 8 to 25, preferably 10 to 21 carbon atoms and R ⁴ and R ^{5 are} identical or different alkyl radicals having 1 to 3 carbon atoms, in particular C ₁₀ -C ₁₈ alkyl dimethylcarboxymethylbetain and C ₁₁ -C ₁₇ -Alkylamidopropyl dimethylcarboxymethyl betaine.

If cationic surfactants are included in the composition, these are preferably the quaternary ammonium compounds of the formula (R ⁶ ) (R ⁷ ) (R ⁸ ) (R ⁹ ) N ⁺ X ^- , in which R ⁶ to R ^{9 are} four identical or different, in particular two long and two short-chain, alkyl radicals and X ^{- are} an anion, in particular a halide ion, for example didecyl-dimethyl-ammonium chloride, alkyl-benzyl-didecylammonium chloride and their mixtures. Preferably, however, the detergent composition is free of cationic surfactants.

When choosing suitable surfactants, care must be taken that they are compatible with the enzymes used. Preferred is enzyme activity for a typical 15 minute median exposure time not reduced by more than 50%, in particular by not more than 30%. Thus, experiments have shown that the known as denaturing sodium dodecyl sulfate (SDS), the activity of the corolase already in a concentration of 1% within 15 minutes to below 30%, while the activity of xylanase even with only 0.1% SDS to less than 40%, so that this surfactant is not preferred for use in cleaning agents according to the invention. If this or a similarly acting surfactant still be used, it is preferable to ensure a spatial separation during storage, for example by encapsulation of the enzymes or by the use of a multi-chamber bottle, so that enzymes and surfactants together until immediately before or during use get in touch. By contrast, alkyl polyglycosides are well suited for use in enzyme-containing compositions according to this invention. Thus, the activity of corolase even with 3% APG 600 remains well over 50%, the xylanase even experiences an increase in activity (more than 150% activity with 0.1% surfactant) with the addition of APG 600 and always shows 3% APG still over 80% activity on.

Farther can the agents according to the invention Builder included. Suitable builders are, for example, alkali metal gluconates, citrates, nitrilotriacetates, carbonates and bicarbonates, in particular Sodium gluconate, citrate and nitrilotriacetate and sodium and potassium carbonate and bicarbonate, and alkali metal and alkaline earth metal hydroxides, especially sodium and potassium hydroxide, ammonia and amines, in particular mono- and triethanolamine, or mixtures thereof. This includes as well the salts of glutaric acid, Succinic acid, adipic acid, tartaric acid and benzene hexacarboxylic acid as well as phosphonates and phosphates. The agents can be builders in quantities to the composition, from 0.1 to 5 wt .-%.

Furthermore, the agents according to the invention may contain acids and / or alkalis. These serve on the one hand as pH regulators, on the other hand, however, the acids can also contribute to the removal of limescale from the surfaces to be cleaned. The acids which can be used according to the invention may be inorganic mineral acids, for example hydrochloric acid, and / or C _1-6 -mono-, di-, tri- or polycarboxylic acids or -hydroxycarboxylic acids such as, for example, formic acid, acetic acid, lactic acid, citric acid, gluconic acid, glutaric acid , Succinic acid, adipic acid, tartaric acid or else malic acid, as well as other organic acids such as salicylic acid or amidosulfonic acid. However, the citric acid is particularly preferably used; also mixtures of several acids can be used. Acids can be present in the inventive detergent in amounts of up to 6 wt .-%, based on the total composition.

To the optionally usable bases include alkanolamines, for example Mono- or diethanolamine, and ammonium or alkali metal hydroxides, especially sodium hydroxide. The cleaning agent according to the invention may contain bases in amounts of up to 2.5% by weight, based on the overall composition.

The composition according to the invention can furthermore contain one or more thickeners for regulating the viscosity. Natural and synthetic polymers and inorganic thickeners are suitable as thickeners. The polymers which can be used include polysaccharides or heteropolysaccharides and other organic natural thickeners, including the polysaccharide gums such as gum arabic, agar, alginates, carrageenans and their salts, guar, guar gum, tragacanth, gellan, Ramzan, dextran or xanthan and their derivatives, eg propoxylated Guar, and their mixtures, as well as pectins, polyoses, carob seed flour, starch, dextrins, gelatin, casein. Furthermore, organic modified natural substances such as carboxymethyl cellulose and cellulose ethers, hydroxyethyl and propyl cellulose and the like or cellulose acetate and core flour ethers can be used. Homogeneous and copolymeric polycarboxylates, especially polyacrylic and polymethacrylic compounds, as well as vinyl polymers, polycarboxylic acids, polyethers, polyimines or else polyamides can serve as organic fully synthetic thickeners. Suitable inorganic thickeners include polysilicic acids, clay minerals such as montmorillonites, zeolites, silicic acid and various nanoparticulate inorganic compounds such as nanoparticulate metal oxides, oxide hydrates, hydroxides, carbonates and phosphates and silicates having an average particle size of 1 to 200 nm, based on the Particle diameter in the longitudinal direction, ie in the direction of the largest expansion of the particles. These nanoparticulate substances may optionally be treated in one further embodiment of the invention with one or more surface modifiers. The surface modification is carried out in known manner to the skilled person with mono- and polybasic carboxylic acids or C _2-8 hydroxycarboxylic acids, functional silanes of the type (OR) _4-n SiR _n (R = org. Radicals having functional groups such as hydroxyl, carboxyl, ester , Amine, epoxy, etc.), quaternary ammonium compounds or amino acids as well as other substances which can be used for this purpose.

In addition to the previously mentioned thickening agents, the inventive cleaning agent Elek containing trolyte salts. These can also contribute to an increase in viscosity. Electrolyte salts in the context of the present invention are salts which decompose into their ionic constituents in the aqueous agent according to the invention. Preference is given to the salts, in particular alkali metal and / or alkaline earth metal salts, of an inorganic acid, preferably of an inorganic acid from the group comprising the hydrohalic acids, nitric acid and sulfuric acid, in particular the chlorides and sulfates. Within the scope of the teaching according to the invention, an electrolyte salt can also be used in the form of its corresponding acid / base pair, for example hydrochloric acid and sodium hydroxide instead of sodium chloride.

in the agents according to the invention can organic and / or inorganic thickener in quantities of total up to 2% by weight, based on the total composition.

The inventive agent can advantageously additionally one or more water-soluble organic solvents usually in an amount of up to 6% by weight, based on the total composition. The solvent is within the scope of the teaching of the invention as required in particular as hydrotrope, viscosity regulator and / or cold stabilizer used. It has a solution-promoting effect especially for Surfactants and electrolyte as well as perfume and dye and so contributes their incorporation at, prevents the formation of liquid-crystalline Phases and contributes to the formation of clear products. The viscosity of the agent according to the invention decreases with increasing amount of solvent. Too much solvent However, it can cause excessive viscosity drop. Finally sinks with increasing amount of solvent the Kältetrübungs- and Clear point of the agent according to the invention.

Suitable solvents are, for example, saturated or unsaturated, preferably saturated, branched or unbranched C _1-20 -hydrocarbons, preferably C _2-15 -hydrocarbons, having at least one hydroxy group and optionally one or more ether functions COC, ie the carbon atom chain interrupting oxygen atoms. However, preferred solvents are the - optionally unilaterally etherified with a C _1-6 alkanol - C _2-6 alkylene glycols and poly-C _2-3 alkylene glycol having an average of 1 to 9 identical or different, preferably identical, alkylene glycol groups per molecule, for example Ethylene glycol, propylene glycol, butylene glycol, diethylene glycol, dimethoxy diglycol, dipropylene glycol, propylene glycol butyl ether, propylene glycol propyl ether, dipropylene glycol monomethyl ether, and PEG. Further preferred solvents are the C _1-6 -alcohols, such as methanol, ethanol, n-propanol, isopropanol, n-butanol, t-butanol, etc., more preferably ethanol and / or isopropanol is used.

When solubilizers can except the solvents described above for example, alkanolamines and alkylbenzenesulfonates with 1 to 3 carbon atoms in the alkyl radical, e.g. Xylene or cumene sulfonate be used.

Further usable hydrotropes are e.g. Octyl sulfate or butyl glucoside. The agent according to the invention can this hydrotrope in amounts of, based on the total composition, contain up to 4 wt .-%.

In an embodiment can the agents according to the invention Containing abrasives. As Abrasivkomponente this can be solid water-soluble and water-insoluble, preferably inorganic compounds and mixtures thereof are used. These include for example, alkali carbonates, alkali bicarbonates and alkali sulfates, Alkaline borates, alkali phosphates, silica, crystalline or amorphous alkali metal silicates and phyllosilicates, fine crystalline sodium aluminum silicates and calcium carbonate. The advantage of water-soluble abrasive components consists in a virtually residue-free rinsability of the agent. Besides these inorganic substances can also Abrasives obtained from animate nature, for example minced nutshells or woods, and abrasion-resistant plastics, for example polyethylene beads, or ceramic or glass beads Find use. The cleaning agent according to the invention can abrasives in amounts of up to 2 wt .-%, based on the total composition.

In addition to the abovementioned components, the compositions according to the invention may contain one or more other auxiliaries and additives, such as are customary, above all, in hard surface cleaners. These include in particular UV stabilizers, corrosion inhibitors, cleaning enhancers, antistatic agents, preservatives (eg 2-bromo-2-nitropropane-1,3-diol or an isothiazolinone Bromnitropropandiol preparation), perfume, dyes, pearlescing agents (for example, glycol distearate) and opacifiers or also skin protection agents, such as those in EP 522 506 are described. The amount of such additives is usually not more than 12 wt .-% in the detergent. The lower limit of the use depends on the nature of the additive and can be up to 0.001 wt .-% and below, for example, in the case of dyes. The amount of auxiliaries is preferably between 0.01 and 7% by weight, in particular 0.1 and 4% by weight.

When Dyes can all in household cleaning products usually used dyes are used. Also with the fragrances can all usual perfumes be used. Preference is given to fruity scents, for example Citrus, pine (Pine) and mint, as well as floral scents. conservation Mitel have a biocidal effect, so that it is desirable in detergents according to the invention only low concentrations, but preferably no preservatives use.

It is advantageous if the agent contains no complexing agent; of the Addition of a bleaching agent for the cleaning agent according to the invention is not required.

Of the pH value of the agents according to the invention is preferably between 1 and 7.5, more preferably between 2 and 5, in particular between 2.5 and 4.5. It is in multiphase Means below the pH of the agent, the pH of the resulting after shaking temporary To understand emulsion, for means offered in multi-chambered bottles the pH of the agent is the pH of the common intended dosage obtained from the components stored in the various chambers Solution. Thus, it is meant in each case the pH of the ready-to-use cleaning solution.

Provided it is a liquid This agent preferably has a viscosity of up to to 1000 mPas. Gel-like or pasty Cleaning agents can In contrast, viscosities of up to 150,000 mPas. The viscosity measurements are carried out at 20 ° C in the Brookfield viscometer LVDV II with a rotor frequency of 20 rpm (spindle no. 31, conc. 100%).

The Detergent may contain one or more propellants (INCI Propellants), usually in an amount of 1 to 80% by weight, preferably 1.5 to 30% by weight, in particular 2 to 10 wt .-%, particularly preferably 2.5 to 8 wt .-%, most preferably 3 to 6 wt .-%, contained.

propellant are customary according to the invention Propellant gases, especially liquefied or compressed gases. The choice depends on the product to be sprayed and the field of application. When using compressed gases such as nitrogen, carbon dioxide or nitrous oxide, in general in the liquid Detergent insoluble are, the operating pressure decreases with each valve actuation. In the detergent soluble or even as a solvent acting liquefied Gases (liquefied gases) as blowing agents offer the advantage of constant operating pressure and even distribution, because in the air the propellant vaporizes and takes up a hundredfold Volume.

Suitable are the following according to INCI Propellants: Butanes, Carbon Dioxide, Dimethyl Carbonate, Dimethyl Ethers, ethanes, hydrochlorofluorocarbon 22, hydrochlorofluorocarbon 142b, hydrofluorocarbon 152a, hydrofluorocarbon 134a, hydrofluorocarbon 227ea, Isobutane, Isopentane, Nitrogen, Nitrous Oxide, Pentane, Propane.

On Chlorofluorocarbons (chlorofluorocarbons, CFCs) as However, propellant is hard because of its harmful effect on the - UV radiation protective - ozone shield the atmosphere, the so-called ozone layer, preferably largely and in particular Completely waived.

preferred Propellants are liquefied gases. Liquid are gases, which are converted at mostly already low pressures and 20 ° C from gaseous to liquid state can. In particular, under liquefied gases but the - in oil refineries as by-products of distillation and cracking of crude oil as well in natural gas processing arising in the gasoline separation - hydrocarbons propane, Propene, butane, butene, isobutane (2-methylpropane), isobutene (2-methylpropene, Isobutylene) and mixtures thereof.

Especially preferably contains the cleaning agent as one or more propellants propane, butane and / or Isobutane, especially propane and butane, most preferably propane, butane and isobutane.

Oral, dental and / or Denture care products

The oral, Tooth and / or dental prosthesis care agents can be used, for example, as mouthwash, Gel, liquid Toothbrush lotion, stiff toothpaste, chewing gum, denture cleaner or Denture adhesive cream are present.

For this It is necessary that the hybrid enzymes according to the invention in a suitable carrier incorporate.

When carrier can e.g. also powdery Preparations or aqueous-alcoholic solutions serve as mouthwashes 0 to 15% by weight of ethanol, 1 to 1.5% by weight of aroma oils and 0.01 to 0.5% by weight sweeteners or as mouthwash concentrates 15 to 60 wt .-% ethanol, 0.05 to 5% by weight of aroma oils, 0.1 to 3% by weight sweeteners and possibly other excipients may contain and before use with water dilute become. The concentration of the components must be chosen so high, that after dilution the mentioned lower concentration limits in the application not be fallen below.

When carrier can but gels as well as more or less flowable pastes serve that out flexible plastic containers or tubes and with the help of a toothbrush on the teeth be applied. Such products contain higher amounts of humectants and binders or consistency regulators and polishing components. Furthermore Aromatic oils, sweeteners and water are also contained in these preparations.

When Humectants can while e.g. Glycerol, sorbitol, xylitol, propylene glycols, polyethenylene glycols or Mixtures of these polyols, especially those Polyethenylenglycole with molecular weights of 200 to 800 (from 400 to 2000) are used be. Preferred as a humectant sorbitol in an amount from 25 - 40 Wt .-% included.

When Anti-calculus agents and as demineralization inhibitors can condensed phosphates in the form of their alkali metal salts, preferably in Form their sodium or Be contained potassium salts. The aqueous solutions of these phosphates react alkaline due to hydrolytic effects. By addition of acid is the pH of the oral, dental and / or dental prosthesis care agents according to the invention adjusted to the preferred values of 7.5-9.

It can also mixtures of different condensed phosphates or hydrated Salts of condensed phosphates are used. The specified Quantities from 2 - 12 However,% by weight refers to the anhydrous salts. Prefers As condensed phosphate is a sodium or potassium tripolyphosphate in an amount of 5 - 10 Wt .-% of the composition.

A preferred active ingredient is a caries-inhibiting fluorine compound, preferably from the group of fluorides or monofluorophosphates in an amount of 0.1 to 0.5 wt .-% fluorine. Suitable fluorine compounds are, for example, sodium monofluorophosphate (Na ₂ PO ₃ F), potassium monofluorophosphate, sodium or potassium fluoride, tin fluoride or the fluoride of an organic amino compound.

Suitable binders and consistency regulators are, for example, natural and synthetic water-soluble polymers such as carrageenan, tragacanth, guar, starch and their non-ionic derivatives such as hydroxypropyl guar, hydroxyethyl starch, cellulose ethers such as hydroxyethyl cellulose or methylhydroxypropyl cellulose. Also, agar-agar, xanthan gum, pectins, water-soluble carboxyvinyl polymers (for example Carbopol ^® types), polyvinyl alcohol, polyvinyl pyrrolidone, high molecular weight polyethylene glycols (molecular weight 10 ³ to 10 ⁶ D). Further substances which are suitable for viscosity control are phyllosilicates such as montmorillonite clays, colloidal thickening silicas, eg airgel silica or fumed silicas.

As polishing components may all heretofore known polishing agent, but preferably precipitated and gel silicas, aluminum hydroxide, aluminum silicate, alumina, alumina trihydrate, insoluble sodium metaphosphate, calcium pyrophosphate, calcium hydrogen phosphate, dicalcium phosphate, chalk, hydroxyapatite, hydrotalcites, talc, magnesium aluminum silicate (Veegum ^®), calcium sulfate, magnesium carbonate, Magnesium oxide, sodium aluminum silicates, such as zeolite A or organic polymers, such as polymethacrylate, are used. The polishing agents are preferably used in smaller amounts of eg 1-10% by weight.

The dental and / or oral care products according to the invention can be improved in their organoleptic properties by adding aroma oils and sweeteners. Aromatic oils are all natural and synthetic flavors which are customary for oral, dental and / or dental care products. Natural flavors can be used both in the form of the essential oils isolated from the drugs and the individual components isolated from them. Preferably, at least one aromatic oil from the group of peppermint oil, spearmint oil, aniseed oil, caraway oil, eucalyptus oil, fennel oil, cinnamon oil, geranium oil, sage oil, thyme oil, marjoram oil, basil oil, citrus oil, Gaultheria oil or one or more synthetic components of these oils isolated therefrom should be included. The most important components of the oils mentioned are, for example, menthol, carvone, anethole, cineole, eugenol, cinnamaldehyde, geraniol, citronellol, linalool, salvos, thymol, terpinene, terpinol, methylchavicol and methyl salicylate. Further suitable flavors are, for example, menthyl acetate, vanillin, ionone, linalyl acetate, rhodinol and piperitone. Suitable sweeteners are either natural ones Sugars such as sucrose, maltose, lactose and fructose or synthetic sweeteners such as saccharin sodium salt, sodium cyclamate or aspartame.

When Surfactants are especially alkyl and / or alkenyl (oligo) glycosides used. Their preparation and use as surface-active Substances are, for example, from US-A-3,839,318, US-A-3,707,535, US-A-3 547 828 DE-A-19 43,689, DE-A-20 36 472 and DE-A-30 01 064 and EP-A-77 167. Regarding the Glycosidrestes holds that both Monoglycosides (x = 1) in which a pentose or hexose residue is glycosidic to a primary Alcohol with 4 to 16 carbon atoms is bound, as well as oligomeric glycosides with a degree of oligomerization x to 10 are suitable. The degree of oligomerization is a statistical one Mean, the one for such technical products usual Homolog distribution is based.

As alkyl and / or alkenyl (oligo) glycoside, an alkyl and / or alkenyl (oligo) glucoside of the formula RO (C ₆ H ₁₀ O) _x -H in which R is an alkyl radical and is preferably suitable / or alkenyl group having 8 to 14 carbon atoms and x has an average of 1 to 4. Particular preference is given to alkyl oligoglucosides based on hydrogenated C _12/14 coconut oil having a DP of from 1 to 3. The alkyl and / or alkenyl glycoside surfactant can be used very sparingly, with amounts of from 0.005 to 1% by weight already being used. are sufficient.

In addition to the said alkyglucoside surfactants, other nonionic, ampholytic and cationic surfactants may also be present, such as: fatty alcohol polyglycol ether sulfates, monoglyceride sulfates, monoglyceride ether sulfates, mono- and / or dialkyl sulfosuccinates, fatty acid isethionates, fatty acid sarcosinates, fatty acid taurides, fatty acid glutamates, ether carboxylic acids, fatty acid glucamides, alkylamido betaine and / or protein fatty acid condensates, the latter preferably based on wheat proteins. In particular, to solubilize the usually water-insoluble aroma oils, a non-ionic solubilizer from the group of surface-active compounds may be required. Particularly suitable for this purpose are, for example, ethoxylated fatty acid glycerides, ethoxylated fatty acid sorbitan partial esters or fatty acid partial esters of glycerol or sorbitan ethoxylates. Solubilizers from the group of ethoxylated Fettsäureglyceride include especially addition products of 20 to 60 moles of ethylene oxide with mono- and diglycerides of linear fatty acids having 12 to 18 carbon atoms or triglycerides of hydroxy fatty acids such as oxystearic acid or ricinoleic acid. Further suitable solubilizers are ethoxylated fatty acid sorbitan partial esters; these are preferably addition products of 20 to 60 moles of ethylene oxide with sorbitan monoesters and sorbitan diesters of fatty acids with 12 to 18 carbon atoms. Also suitable solubilizers are fatty acid partial esters of glycerol or sorbitan ethoxylates; these are preferably mono- and diesters of C ₁₂ -C ₁₈ -fatty acids and adducts of 20 to 60 moles of ethylene oxide with 1 mole of glycerol or 1 mole of sorbitol.

The oral, Tooth and / or denture care agents preferably contain as a solubilizer for possibly contained aroma oils Addition products of 20 to 60 moles of ethylene oxide to hardened or uncured castor oil (i.e., oxystearic acid or ricinoleic acid triglyceride), on glycerol mono- and / or distearate or sorbitan mono- and / or distearate.

• – Pigmente, z.B. Titandioxid, und/oder Farbstoffe
• – pH-Stellmittel und Puffersubstanzen wie z.B. Natriumbicarbonat, Natriumcitrat, Natriumbenzoat, Zitronensäure, Phosphorsäure oder saure Salze, z.B. NaH₂PO₄
• – wundheilende und entzündungshemmende Stoffe wie z. B. Allantoin, Harnstoff, Panthenol, Azulen bzw. Kamillenextrakt
• – weitere gegen Zahnstein wirksame Stoffe wie z.B. Organophosphonate, z.B. Hydroxyethandiphosphonate oder Azacycloheptandiphosphonat
• – Konservierungsstoffe wie z.B. Sorbinsäure-Salze, p-Hydroxybenzoesäure-Ester.
• – Plaque-Inhibitoren wie z.B. Hexachlorophen, Chlorhexidin, Hexetidin, Triclosan, Bromchlorophen, Phenylsalicylsäureester.

Other common additives for oral, dental and / or denture care medium are, for example

• - Pigments, such as titanium dioxide, and / or dyes
• PH adjusting agents and buffer substances, for example sodium bicarbonate, sodium citrate, sodium benzoate, citric acid, phosphoric acid or acid salts, for example NaH ₂ PO ₄
• - wound healing and anti-inflammatory substances such. Allantoin, urea, panthenol, azulene or chamomile extract
• - Other anti-calculus substances such as organophosphonates, eg Hydroxyethandiphosphonate or Azacycloheptandiphosphonat
• - Preservatives such as sorbic acid salts, p-hydroxybenzoic acid esters.
• - Plaque inhibitors such as hexachlorophene, chlorhexidine, hexetidine, triclosan, bromochlorophene, Phenylsalicylsäureester.

In a particular embodiment the composition is a mouthwash, a mouthwash, a denture cleanser or a denture adhesive.

For inventively preferred Prosthesis cleaners, in particular prosthesis cleansing tablets and powder, are in addition to the ingredients already mentioned for the oral, Dental and / or denture care additionally per-compounds such as for example peroxoborate, peroxomonosulfate or percarbonate. she have the advantage of being next to the bleaching effect at the same time also deodorizing and / or disinfecting act. The use of such Per compounds in denture cleaners is between 0.01 and 10 wt .-%, in particular between 0.5 and 5 wt .-%.

When other ingredients are also enzymes, e.g. Proteases and carbohydrase, suitable for breaking down proteins and carbohydrates. The pH can be between pH 4 and pH 12, in particular between pH 5 and pH 11 lie.

For the prosthesis cleansing tablets, additional auxiliaries are additionally necessary, such as, for example, agents which give off a bubbling effect, for example CO ₂ releasing substances such as sodium bicarbonate, fillers, for example sodium sulfate or dextrose, lubricants, for example magnesium stearate, flow regulators, for example colloidal silica and granulating agents, such as the already mentioned high molecular weight polyethylene glycols or polyvinylpyrrolidone.

Denture adhesives can be offered as powders, creams, foils or liquids and support the adhesion of the prostheses.

When Active ingredients are natural and synthetic swelling substances. As natural swelling substances are in addition Alginates also include plant gums such as e.g. Gum arabic, tragacanth and karaya gum as well as more natural To take rubber. In particular, alginates and synthetic have Swelling agents, e.g. Sodium carboxymethylcellulose, high molecular weight Ethylene oxide copolymers, salts of poly (vinyl ether-co-maleic acid) and Polyacrylamides.

When Excipients for pasty and liquid Products are particularly suitable hydrophobic basics, in particular Hydrocarbons, such as white Vaseline (DAB) or paraffin oil.

pictures

1 schematically shows the DNA fragment of glucanase (Glu), 5 shows the associated DNA and protein sequences (SEQ ID NO: 3 and 4) (see Example 1).

2 schematically shows the DNA fragment of the hybrid enzyme from glucanase and small binding domain (KBD), 6 shows the associated DNA and protein sequences (SEQ ID NO: 7 and 8) (see Example 2).

3 schematically shows the DNA fragment of the hybrid enzyme from glucanase and large binding domain (GBD), 7 shows the associated DNA and protein sequences (SEQ ID NO: 12 and 13) (see Example 3).

4 schematically shows the DNA fragment of the hybrid enzyme from glucanase and middle binding domain (MBD), 8th shows the associated DNA and protein sequences (SEQ ID NO: 17 and 18) (see Example 4).

embodiments

Example 1:

Isolation of the β-glucanase gene (EC 3.2.1.6, X00754) from B. subtilis

DNA was isolated by standard methods with the DNeasy Kit (Qiagen) from B. subtilis. The gene was amplified in a PCR reaction (1 × PCR buffer, Gibco, 0.2 mM per dNTP, Gibco, 1.5 mM MgCl ₂ , Gibco, 2.5 U Taq polymerase, Gibco) with the primers GluBcI ( 5'TGATCATGCCTTATCTGAAACGAG3 ') (SEQ ID NO: 1) and GIuSac (5'GAGCTCTTATTTTTTTGTATACGCC3') (SEQ ID NO: 2) (both each 0.6 μM) in a thermocycler (Eppendorf) under the following conditions:
94 ° C 5 minutes
30 cycles:
94 ° C for 30 seconds
55 ° C 30 seconds
72 ° C 1 minute 30 seconds
final elongation:
72 ° C 7 minutes

The Amplificate was cloned into the TA cloning vector (pGEM-Teasy, Promega) cloned and in XL1 Blue MRF'E. coli cells (Stratagene) transformed (according to the manufacturer's instructions).

For the expression the isolated gene had to be converted into a Bacillus expression system. The plasmid was purified using the QIAprep Spin Miniprep Kit (Qiagen) from an overnight culture isolated from the transformed E. coli cells. Subsequently was the gene with the restriction enzymes BcII and SacI from the pGEM-teasy Vector cut and placed in the appropriate interfaces of the Henkel Bacillus expression vector pAWA31 integrated. The transformed Bacillus subtilis cells were glucanase deficient prior to transformation (strain MW 10, Bacillus Genetic Stock Center, Columbus, USA).

Example 2:

Isolation of the β-glucanase (EC 3.2.1.6, X00754) from B. subtilis incorporating the small positive binding domain (KBD):

The small binding domain corresponds to the DNA region 238-276 base pairs from the sbsA gene (X71092) from Geobacillus stearothermophilus. The gene encodes for in surface protein. The sequence was by extension the primer of glucanase (without stop codon) added. The Stop codon was placed behind the region of the sbsA gene.

DNA was isolated by standard methods with the DNeasy Kit (Qiagen) from B. subtilis. The gene was amplified in a PCR reaction (1 × PCR buffer, Gibco, 0.2 mM per dNTP, Gibco, 1.5 mM MgCl ₂ , Gibco, 2.5 U Taq polymerase, Gibco) with the primers GluBcI ( 5'TGATCATGCCTTATCTGAAACGAG3 ') (SEQ ID NO: 5) and GIuKyrSac (5'GAGCTCCTAACGGTATCGTTTTTTCGCTTTGTTGTATTCAGCATATACTTTTTTTGTAT AGCGC3') (SEQ ID NO: 6) (both each 0.6 μM) in a thermocycler (Eppendorf) under the following conditions:
94 ° C 5 minutes
30 cycles:
94 ° C for 30 seconds
55 ° C 30 seconds
72 ° C 50 seconds
final elongation:
72 ° C 7 minutes

The Amplificate was cloned into the TA cloning vector (pGEM-Teasy, Promega) cloned and in XL1 Blue MRF'E. coli cells (Stratagene) transformed (according to the manufacturer's instructions).

For the expression the isolated gene had to be converted into a Bacillus expression system. The plasmid was purified using the QIAprep Spin Miniprep Kit (Qiagen) from an overnight culture isolated from the transformed E. coli cells. Subsequently was the gene with the restriction enzymes BcII and SacI from the pGEM-teasy Vector cut and placed in the appropriate interfaces of the Henkel Bacillus expression vector pAWA31 integrated. The transformed Bacillus subtilis cells were glucanase deficient prior to transformation (strain MW10, Bacillus Genetic Stock Center, Columbus, USA).

Example 3:

Isolation of the β-glucanase (EC 3.2.1.6, X00754) from B. subtilis incorporating the large positive binding domain (GBD):

The size positive binding domain corresponds to the DNA region 276-686 base pairs from the sbsC gene (AF055578) from Geobacilus stearothermophilus (DSM22). The gene encodes a surface protein. The Sequence was isolated by PCR and molecular biology at the C-terminus of glucanase ligated. The ligation was carried out by a Kas interface, which works as a linker between the two domains.

DNA was prepared by standard methods with the DNeasy Kit (Qiagen) from B.subtilis and G. stearothermophilus.

The glucanse gene without stop codon was isolated in a PCR reaction from B.subtilis DNA (1 x PCR buffer, Gibco; 0.2 mM per dNTP, Gibco; 1.5 mM MgCl ₂ , Gibco; 2.5 U Taq polymerase, Gibco) with the primers GIuBcI (5'TGATCATGCCTTATCTGAAACGAG3 ') and GluKas (5'GCCTTTTTTGTATAG CGCACCC3') (SEQ ID NO: 9) (both each 0.6 μM) amplified in a thermocycler (Eppendorf) under the following conditions:
94 ° C 5 minutes
30 cycles:
94 ° C for 30 seconds
60 ° C for 30 seconds
72 ° C 50 seconds
final elongation:
72 ° C 7 minutes

The gene segment from the sbsC gene from G. stearothermophilus DNA was analyzed in a PCR reaction (1 × PCR buffer, Gibco, 0.2 mM per dNTP, Gibco, 1.5 mM MgCl ₂ , Gibco, 2.5 U Taq). Polymerase, Gibco) with the primers KassbsC (5'GGCGCCGACAAAAAGAAAGCAGTCAAA3 ') (SEQ ID NO: 10) and SacsbsC (5'GAGCTCTTAGCGCATTTTGTCTAATTTTG3') (SEQ ID NO: 11) (both 0.1 μM each) in a thermocycler (Eppendorf) amplified under the following conditions:
94 ° C 10 minutes
30 cycles:
94 ° C for 30 seconds
60 ° C for 30 seconds
72 ° C 45 seconds
final elongation:
72 ° C for 10 minutes

By the primer SacsbsC became a stop codon at the end of the sbsC section inserted.

The Fragments were made according to manufacturer's instructions in the pGEM-Teasy vector (Promega) and transformed into JM110 cells (Stratagene).

plasmids were isolated using the QiaPrep Spin Kit (Qiagen).

Fragment Isolation

glucanase: The glucanase was isolated from the vector in two steps. Plasmid DNA was mixed with buffer A (final concentration 1 ×) and 50 units of Narl (both Roche) cleaved for 6 h at 37 ° C. The DNA was concentrated with the PCR Purification Kit (Qiagen) and subsequently with 1 × buffer M and 50 units BcII (Roche) cleaved at 37 ° C for 3 h.

SbsC: Gene fragment: The sbsC gene fragment was out in two steps isolated vector. Plasmid DNA was treated with buffer A (final concentration 1 ×) and 50 Units Narl (both Roche) for 3 h at 37 ° C split and then with additional 25 units SacI (Roche) at 37 ° C for 3 h split.

Both Fragments were made on a preparative, 2% agarose gel separated from the vector, excised under UV light and purified from the gel with QIAquick Gel Extraction Kit (Qiagen).

Approximately 20 ng of glucanase were incubated with approximately 60 ng GBD in the presence of 1 × ligation buffer and 0.5 U T4 ligase (Roche) Night at 4 ° C ligated.

The ligation product was incubated with the primers GIuBcI and SacsbsC (both 0.6 μM each) in the presence of (1 × PCR buffer, Qiagen, 0.3 mM per dNTP, Gibco, 2.5 U ProofStart DNA polymerase, Qiagen) a thermal cycler (Eppendorf) under the following conditions:
94 ° C 5 minutes
30 cycles:
94 ° C for 30 seconds
60 ° C for 30 seconds
72 ° C 1 minute 10 seconds
final elongation:
72 ° C 7 minutes

The amplificate was integrated into the Henkel Bacillus expression vector pAWA31 via the built-in BcII and SacI cleavage sites. The transformed Bacillus subtilis cells were before transformation Glucanase Deficient (strain MW10, Bacillus Genetic Stock Center, Columbus, USA).

Example 4:

Isolation of the β-glucanase (EC 3.2.1.6, X00754) from B. subtilis incorporating the middle, positive binding domain (MBD):

at the middle positive binding domain it is an artificial one synthesized DNA section, which is characterized in that it contains regions with positive charges. The synthesis of the MBD took place via two successive PCR reactions. The sequence is going through determines the choice of the primer used. The primers themselves served as template DNA.

DNA was prepared by standard methods with the DNeasy Kit (Qiagen) from B.subtilis isolated.

The glucanse gene without stop codon was isolated in a PCR reaction from B.subtilis DNA (1 × PCR buffer, Gibco; 0.2 mM per dNTP, Gibco; 1.5 mM MgCl ₂ Gibco; 2.5 U Taq Polymerase, Gibco) with the primers GIuBcI (5'TGATCATGCCTTATCTGAAACGAG3 ') and GluKas (5'GGCGCCTTTTTTGTATAG CGCACCC3') (both 0.6 μM each) in a thermocycler (Eppendorf) under the following conditions:
94 ° C 5 minutes
30 cycles:
94 ° C for 30 seconds
60 ° C for 30 seconds
72 ° C 50 seconds
final elongation:
72 ° C 7 minutes

The MBD was synthesized in two consecutive PCR reactions. In the first reaction, an 81 bp fragment was amplified in a thermocycler (Eppendorf) under the following conditions:
1X PCR buffer, Gibco; 0.2mM per dNTP, Gibco; 1.5 mM MgCl ₂ , Gibco; 2.5 U Taq Polymerase, Gibco Primer:
PBDvorne
(5'GGCGCCCAGCTGAAGAAGAAGCTGCAGGCACTCAAGAAGAAGAATGCGCAGCT A3 ') (SEQ ID NO: 14)
PBDmitte
(5'CTTCTTAAGTGCCTGCAGCTTCCACTTTAGCTGCGCATTCTTCTTCTTGA3 ') (SEQ ID NO: 15) (both 0.1 μM each)
PCR scheme:
94 ° C 5 minutes
10 cycles:
94 ° C for 30 seconds
60 ° C for 30 seconds
72 ° C for 10 seconds
final elongation:
72 ° C 7 minutes

The 81 bp fragment was transferred to a new PCR reaction (1X PCR buffer, Gibco, 0.2 mM dNTP, Gibco, 1.5 mM MgCl ₂ , Gibco, 2.5 U Taq polymerase, Gibco) Primer PBDfront
(5'GGCGCCCAGCTGAAGAAGAAGCTGCAGGCACTCAAGAAGAAGAATGCGCAGCT A3 ') and PBDins
(5'GAGCTCTCAACAACCGCCCTGCGCCAGCTTCTTCTTAAGTGCCTGCAGCT3 ') (SEQ ID NO: 16) (both 0.1 μM each). The amplification scheme was as follows:
94 ° C 5 minutes
10 cycles:
94 ° C for 30 seconds
60 ° C for 30 seconds
72 ° C for 10 seconds
final elongation:
72 ° C 7 minutes

By the primer PBD backbone was a stop codon at the end of the PBD section inserted.

The Fragments were made according to manufacturer's instructions in the pGEM-Teasy vector (Promega) and transformed into XL1-Blue MRF 'cells (Stratagene).

plasmids were isolated using the QiaPrep Spin Kit (Qiagen).

Fragment Isolation

glucanase: The glucanase was isolated from the vector in two steps. Plasmid DNA was mixed with buffer A (final concentration 1 ×) and 50 units of Narl (both Roche) cleaved for 6 h at 37 ° C. The DNA was concentrated with the PCR Purification Kit (Qiagen) and subsequently with 1 × buffer M and 50 units BcII (Roche) cleaved at 37 ° C for 3 h.

MBD: The MBD fragment was isolated from the vector in two steps. Plasmid DNA was mixed with buffer A (final concentration 1 ×) and 50 units of Narl (both Roche) for 3 h at 37 ° C split and connecting with additional 25 Unit SacI (Roche) at 37 ° C for 3 h split.

Both Fragments were made on a preparative, 2% agarose gel separated from the vector, excised under UV light and purified from the gel with QIAquick Gel Extraction Kit (Qiagen).

Approximately 20 ng of the glucanase were mixed with approximately 60 ng of MBD in the presence of 1 × ligation buffer and 0.5 U T4 ligase (Roche) Night at 4 ° C ligated.

The ligation product was incubated with the primers GIuBcI and PBDhinten (both 0.6 μM each) in the presence of (1 × PCR buffer, Qiagen, 0.3 mM per dNTP, Gibco, 2.5 U ProofStart DNA polymerase, Qiagen) a thermal cycler (Eppendorf) under the following conditions:
94 ° C 5 minutes
30 cycles:
94 ° C for 30 seconds
60 ° C for 30 seconds
72 ° C 1 minute 10 seconds
final elongation:
72 ° C 7 minutes

The Amplificate was over the built-in BcII and SacI sites in the Henkel Bacillus expression vector pAWA31 integrated. The transformed Bacillus subtilis cells were glucanase deficient prior to transformation (strain MW10, Bacillus Genetic Stock Center, Columbus, USA).

Example 5:

A B. subtilis glucanase was cloned. This enzyme was genetically engineered with three different, positively charged binding domains:
GIuMBD: glucanase + linker + artificial binding domain
GIuKBD: Glucanase + small positively charged area of surface protein from G. stearothermophilus
GIuGBD: glucanase + linker + large positively charged area of surface protein from G. stearothermophilus

For checking the function of the cloned enzymes, supernatants from overnight cultures were used in each case and adjusted to comparable enzyme activities ( 1 ).

The Binding and hydrolysis ability This cloned enzyme was compared to unmodified glucanase tested on pectin agar (negatively charged polysaccharide). the Agar was insoluble Glucan added. The cloned enzymes became activity-identical used. Each cloned enzyme was incubated at 4 ° C for 5 minutes Agar pipetted, removed and washed twice with buffer. Of the Agar was then overlaid with buffer and at 37 ° C incubated. It was shown by a blue color that only where hybrid enzymes (GIuMBD, GIuKBD, GIuGBD) were pipetted onto the agar, the insoluble glucan was hydrolyzed.

Example 6:

Example formulations for household cleaners

SEQUENCE LISTING

Claims (29)

Hybrid enzyme comprising a hydrolytically active Unit and a cationic binding unit. Hybrid enzyme according to claim 1, characterized that it is a fusion protein. Hybrid enzyme according to claim 1 or 2, characterized that the hydrolytically active unit is a peptide or protein that is capable of oligo- or polysaccharides dismantle. Hybrid enzyme according to claim 1 or 2, characterized that the hydrolytically active unit is a peptide or protein that is capable of oligo- or polypeptides dismantle. Hybrid enzyme according to claim 1 or 2, characterized that the hydrolytically active unit is a peptide or protein that is capable of oligo- or polynucleotides dismantle. Hybrid enzyme according to one of claims 1 to 5, characterized the cationic binding unit is a polypeptide of up to 150 amino acids includes. Hybrid enzyme according to one of claims 1 to 6, characterized that the cationic binding unit averages between 0.01 and 0.8 positive charges per amino acid residue at neutral pH having. Hybrid enzyme according to one of claims 1 to 7, characterized that the cationic binding unit is an artificial one produced sequence is. Hybrid enzyme according to one of the claims 8, characterized in that that the cationic binding unit is a polypeptide with an amino acid sequence according to SEQ ID NO: 21 or a derivative thereof. Hybrid enzyme according to one of claims 1 to 7, characterized that the cationic binding domain is a polypeptide fragment a surface layer protein or a derivative thereof. Hybrid enzyme according to claim 10, characterized that the cationic binding unit is a polypeptide with an amino acid sequence according to SEQ ID NO: 19 or according to SEQ ID NO: 20 or a derivative of said polypeptides. An enzyme preparation comprising a hybrid enzyme one of the claims 1 to 11. Enzyme preparation according to claim 12, characterized in that that in addition unmodified enzymes selected from polysaccharidases, proteases and nucleases. Preparation selected from the group consisting from sterilization, disinfection, washing and cleaning agents, characterized in that it is a hybrid enzyme according to any one of claims 1 to 11 or an enzyme preparation according to claim 12 or 13. Preparation according to claim 14, characterized that the hybrid enzyme and / or the enzyme preparation in amounts up to to 10 wt .-% in the preparation are included. Preparation according to claim 14 or 15, characterized that it contains at least one further component selected from the group consisting from surfactants, builders, co-builders, acids, alkalis, thickeners, solvents solubilizers and abrasives. Preparation according to one of claims 14 to 16, characterized that it contains at least one other excipient or additive selected from the group consisting of UV stabilizers, corrosion inhibitors, Cleaning amplifiers Antistatic agents, perfumes, Dyes, pearlescing agents, opacifiers and skin protection agents. Product containing an enzyme preparation or a cleaning agent according to any one of the preceding claims and a spray dispenser for dosing the enzyme preparation or the cleaning agent as an aerosol and / or as a foam. Process for the hydrolysis of biofilms on surfaces Incubation with a hybrid enzyme, an enzyme preparation or a Cleaning agent according to one of the preceding claims, optionally under Use of a product according to the preceding claim, characterized that characterized the vitality the microbial cells are not sustainably damaged. Process for the hydrolysis of biofilms on surfaces, characterized characterized in that the hybrid enzyme, the enzyme preparation or the cleaning agent according to any one of the preceding claims, optionally using a product according to the invention according to claim 18, on which biofilm-coated area is applied, optionally with a cloth, a sponge, a brush or any other utensil suitable for cleaning on the Biofilm infested surface is blurred or rubbed and, if necessary after a Exposure time of 1 to 30 minutes, a few hours or overnight, with rinsed with clear water will, whereupon the surface wiped with a dry cloth. Use of hybrid enzymes, enzyme preparations or cleaning agents according to any one of the preceding claims destruction and removal of biofilms on surfaces. Use according to claim 21, characterized that it is at the surface around a hard surface is. Use of hybrid enzymes, enzyme preparations or cleaning agents according to any one of the preceding claims Cleaning of textiles. Product of an enzyme preparation or a cleaning agent according to one of the preceding Claims and a multi-chamber bottle. Cosmetic and / or pharmaceutical preparation containing hybrid enzymes according to any one of claims 1 to 11. Cosmetic and / or pharmaceutical preparation according to the preceding claim, characterized in that it is in the preparation of oral, dental or denture care products is. Use of hybrid enzymes according to any one of claims 1 to 11 for the preparation of a cosmetic preparation. Use of hybrid enzymes according to any one of claims 1 to 11 for the manufacture of a medicament. nucleic acids coding for a polypeptide according to SEQ ID NO: 21 or a derivative thereof.