DE10304331B4 - Enzymatic removal of biofilms on household surfaces - Google Patents

Abstract

Use of a biocidal detergent for hard surfaces, which contains one or more surfactants and for destruction and removal of biofilms 0.1 to 10 wt .-% of an enzyme preparation of at least two enzymes, wherein the at least two enzymes to a polysaccharidase with a Protease or a polysaccharidase with a nuclease, for the destruction and removal of biofilms on household surfaces.

Description

The invention relates to household cleaners with enzymes that are capable of destroying biofilms and in this way to clean hard household surfaces of bacterial deposits without the need for additional biocides.

Biofilm refers to the growth of surfaces by microbes and, above all, the gelatinous protective layer that they collectively deliver and envelop. At the same time, this mucus is the reason why bacteria living under real conditions, as well as other biofilm-producing microorganisms, are much more resistant to - even biocide-containing - cleaning agents than laboratory cultures, which usually consist of individual free-floating bacteria, so-called planktonic cells. Biofilms lead to massive impairments in technical systems and can cause persistent infections in medicine, which are largely resistant to conventional antibiotics. In the household, the biofilm-induced impairments are both hygienic and aesthetic.

The control of biofilms by biocides is only possible to a limited extent because the mucus matrix, which mainly consists of polysaccharides, protects the cells against the action of the biocides (so-called pseudoresistance). In order to achieve any effect, the biocides would therefore have to be used in much higher doses than would be necessary for a comparable amount of unassociated bacteria present. The use of biocides in the home, however, is unfavorable from an ecological point of view, so that the efforts are more likely to provide detergents with only low concentrations or completely without biocides. So far, however, this has not been reconciled with the desire for as complete a removal as possible of bacterial biofilms - and thus optimal hygiene.

The European patent EP 0 946 207 B1 (Corr. WO 98/26 807 A1 ) teaches the enzymatic treatment of biofilms with combinations of hydrolases which serve to detach and remove 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 0 388 115 A1 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 the WO 01/09 295 A1 enzymes with a β- (1,6) -endoglucanase activity and corresponding DNA sequences are claimed as well as an enzyme preparation for the degradation of β-1,6-glucan-containing materials and their use. The β- (1,6) -endoglucanases also serve to remove biofilms from various surfaces. Mainly, however, this Spanish paper is concerned with the degradation of the β-1,6-glucan-rich cell wall of certain fungi.

Also the WO-PS 96/36 569 A1 describes an enzymatic method for removing biofilms. In this case, a mannanase, optionally in combination with at least one other enzyme, is used to prevent slime formation in industrial water-bearing systems and to remove existing mucus. According to WO 96/17632 A1 This object can also be achieved with combinations of short-chain glycol components with at least one enzyme from the group of polysaccharidases, proteases, lipases and glycoproteases.

The international application WO 01/53 010 A1 also teaches an enzymatic process for removing biofilms from various surfaces flushed with water or aqueous solutions in industrial systems as well as in the medical field, wherein the biofilms are both above the water surface, ie at solid / liquid / air interfaces, and under water, at solid / liquid interfaces. The enzymes used are carbohydrases or proteases, combinations of these two enzymes are also possible. Optionally, the cleaning solution in addition to the enzymes mentioned other active ingredients such as corrosion inhibitors, surfactants, biocides, etc. included.

The systems described in the literature all combat biofilms in industrial installations and / or on soft surfaces. In addition, their effectiveness has always been tested only on biofilms in the laboratory were bred and formed only by a few selected bacterial species, so that these systems are only partially transferable to real conditions. The object of this invention is to provide detergents which, without the use of biocides, are able to dissolve biofilms on hard household surfaces and to remove the film-forming microorganisms.

The composition of bacterial biofilms on household surfaces has so far been little researched; The few research findings do not allow any indication of any differences between biofilms on industrial and residential surfaces. Accordingly, the assessment of the effectiveness of a drug was problematic and the comparability of individual results by no means always given.

Therefore, first attempts were made to characterize WC biofilms. From the results obtained, model biofilms were then grown from several microorganisms that more accurately reflect the actual ratios on household surfaces than the biofilms previously studied most commonly from individual bacterial strains available in the laboratory. With these model biofilms, a reliable test system was now available, which is suitable for testing the effectiveness of detergents with or without biocides against biofilms on hard household surfaces.

These model biofilms were then exposed to the action of various enzymes to study degradation; It was surprisingly found that even the effect of polysaccharidases and / or proteases is sufficient to break up the biofilm and detach the cells. These can then simply be rinsed off, so that, in contrast to the methods for removing biofilms described in the prior art, no bactericidal oxidoreductases or stronger biocides are needed. It is therefore an environmentally friendly method for cleaning and disinfecting especially wet, wasserumspülter surfaces have become available in the household.

As has recently been described (C.W. Whitchurch et al., 2002, Science 295, p. 1487), bacterial biofilms also consist of nucleic acids and are particularly useful in early stages by nucleic acid degrading enzymes, e.g. B. hydrolyzed by DNAses.

Such enzymes (nucleases) are thus also suitable for use in a preparation according to the invention.

As described, it has surprisingly been found that one or more enzymes from the group polysaccharidase (β-glucanase, cellulase, xylanase, amylase, dextranase, glucosidase, galactosidase, pectinase, chitinase, lysozyme, alginate lyase) and protease (subtilisin, thermolysin, Pepsin, carboxypeptidase, acid protease) and / or nuclease (DNAse, RNAse) already destroy the bacterial mucus matrix in biofilms on WCs and other temporarily wetted household surfaces, so that the bacteria are released from their dressing and transferred to the planktonic form, thereby providing a simple Rinse sufficient to remove the microbes, without the additional biocides would be needed.

The subject of this application is therefore the use of a biocide-free cleaning agent which contains one or more surfactants and 0.1 to 10 wt .-% of an enzyme preparation of at least two enzymes, wherein the at least two enzymes to a polysaccharidase with a protease or a Polysaccharidase with a nuclease acts to destroy and remove biofilms on household surfaces.

The enzyme preparation is a system of one or more enzymes from the group polysaccharidase (β-glucanase, cellulase, xylanase, amylase, dextranase, glucosidase, galactosidase, pectinase, chitinase, lysozyme, alginate lyase) and / protease (subtilisin , Thermolysin, pepsin, carboxypeptidase, acid protease) and / or nuclease (DNAse, RNAse). This preparation is added to a hard surface cleaner which, in addition to removing further soiling, is also capable of effectively removing biofilms.

In addition to the enzyme preparation, the cleaning agent may also contain surfactants, builders, acids, alkalis, hydrotropes, solvents, thickeners, dyes, perfumes, corrosion inhibitors, skin protection agents and other ingredients customary in detergents. If the detergent is to be sprayed, it may also be possible for a propellant to be present. The cleaning agent is preferably a liquid aqueous agent, but it may also be present as a gel, as a paste or as a powder.

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), The Cosmetic, Toiletry and Fragrance Association (CTFA), 1101 17th Street, NW, Suite 300, Washington, DC, 20036, USA , which publishes more than 9,000 INCI names and references to more than 37,000 trade names and technical names, including related distributors from over 31 countries. 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 indication CAS means that the following sequence of numbers is a name of the Chemical Abstracts Service.

enzyme preparation

The cleaning agent used according to the invention contains an enzyme preparation of at least two enzymes, wherein the at least two enzymes are a polysaccharidase with a protease or a polysaccharidase with a nucleic acid-degrading enzyme.

Polysaccharidases are enzymes which catalyze the hydrolytic cleavage of polysaccharides. For example, cellulose is broken down by the cellulase, whereas glucosidase from the oligo- or polysaccharide molecule splits off individual glucose units from the end. The extracellular polysaccharide matrix of the biofilm consists of homo- and heteropolysaccharides, which are mainly composed of glucose, fucose, mannose, galactose, fructose, pyruvate, mannuronic acid and glucuronic acid. Accordingly, the polysaccharides are, for example, levans, polymannans, dextrans, cellulose, amylopectin, glycogen or alginate.

For the purposes of this invention, all polysaccharidases can be used for the hydrolysis of biofilms. However, the enzyme preparation preferably contains at least one enzyme from the group β-glucanase, cellulase, xylanase, amylase, dextranase, glucosidase, galactosidase, pectinase, chitinase, lysozyme and alginate lyase. These enzymes may each comprise several family members. Thus, for example, both an α-1,4-glucosidase, which was also referred to as maltase, and a β-1,4-glucosidase are known. For the purposes of this invention, all these subgroups are included. Commercially available mixtures of polysaccharidases can also be used. Thus, for example, Viscozyme, a liquid enzyme preparation from Aspergillus sp., Which is composed of various polysaccharidases, u. a. from arabanase, cellulase, β-glucanase, hemicellulase and xylanase, for use in the context of this invention.

Proteases are enzymes that catalyze the hydrolytic cleavage of the peptide bond of proteins and peptides. They can therefore also be used for the degradation of biofilms, since these can contain polysaccharides as well as proteins secreted by the film-forming organisms. Proteases can be systematically classified according to the pH optimum of their action into acidic, neutral and alkaline proteases, but they are classified according to the catalytically active group in the active site. 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, but preferably at least one protease is selected from the group subtilisin, thermolysin, pepsin, carboxypeptidase, acid protease.

The nucleases are nucleic acid degrading enzymes. Depending on the site of hydrolysis, a distinction is made between endonucleases which cleave within the nucleic acid strand (eg pancreatic DNAse I) and exonucleases which release nucleotides from the end (eg exonuclease III from E. coli). DNA single strands (S1 nuclease from Aspergillus oryzae) or double strands (exonuclease III) can serve as a substrate and can be degraded quite unspecifically to shorter strands or individual nucleotides. Mixed specificities are possible and occur depending on z. Of enzyme concentration and presence of certain ions. Similarly, the cleavage of RNA molecules is possible. Highly specific endonucleases require particular nucleotide sequences (typically 4-8 base pairs) and are sometimes referred to as restriction enzymes or restriction endonucleases. Because biofilms are also made Nucleic acids can also exist, all nucleases are suitable for use according to the invention in biofilm degrading agents. Preferably, however, non-specific DNAses or RNAses are used.

The enzymes which can be used for the purposes of this invention can be obtained by conventional biochemical methods from the corresponding organisms. But they are also commercially available, for example from the companies Novozymes, Genencor International, Sigma, AB Enzymes or ASA enzymes. The enzyme preparations may also be of technical quality and consist of several different enzyme activities. The presence of side activities, some of which are not further characterized, can also have a favorable influence on the hydrolysis of the complex biofilm polymers.

If proteases are to be combined with other enzymes, for example polysaccharidases, then it should be noted that these could be attacked in liquid agents by components contained in the cleaner, such as surfactants or organic acids, or else by the rather nonspecifically acting proteases, thereby increasing the effectiveness of the Reduce the mean of non-protein biofilm constituents. To ensure the activity and stability of such detergents over a longer period of time, it is therefore useful to keep sensitive enzymes separate from denaturing components and in particular also proteases from other enzymes, so that these components only meet one another immediately before or during the cleaning process.

This separation of the different enzymes can be done in different ways. One possibility is to introduce the enzymes in microcapsules, the immediately before or during the cleaning process, for example by mechanical action during the cleaning process, by sudden change in the osmotic conditions z. B. when diluting with water or by cutting the capsules when dosing from the storage bottle, open and release their contents. However, it should be noted that the capsule wall in turn must not be attacked by the enzymes. Capsule materials based on proteins or polysaccharides should therefore be excluded. Alternatively, the enzymes can each be immobilized on a solid carrier, which is suspended in the detergent and can additionally serve as an abrasive in the application of the agent. The immobilization is carried out in a manner known to those skilled in the art; as support materials, it is also possible to use polymers in addition to inorganic substances, for example silicates or porous glass.

Another conceivable possibility could also be the incorporation into a multiphase agent, so that the protease would be predominantly located in one phase and the remaining enzymes predominantly exist in a further phase. Such an agent would be shaken shortly before use and the resulting emulsion applied to the area to be cleaned; the agent remaining in the storage bottle would then undergo a rapid phase separation so that the enzymes only come in contact for a short time and possibly at the phase interface. Similarly, the case would be stored if an enzyme were incorporated into a phase which would be stably dispersed in the form of droplets in the second phase containing the other enzyme; In this case, the agent would 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, consumer acceptance would probably be low.

It makes more sense, therefore, to store the two enzymes in spatially separated solutions. The storage in two completely separate bottles is the less preferred solution, as this means a greater effort for the consumer. Therefore, the provision in a two-chamber bottle is particularly preferred, from which then both cleaning solutions can be dosed simultaneously. Such two-chamber bottles are already known from the prior art. Besides the possibility of separately storing mutually incompatible enzymes, a two- or multi-chamber bottle offers further advantages. It is thus possible, inter alia, to store an enzyme in a solution with optimum pH in terms of storage stability in one chamber and to store a solution in another chamber, the pH of which causes a cleaning solution to form when mixing it with the enzyme storage solution, their pH causes an optimum of enzyme activity. For example, many polysaccharidases are particularly stable at neutral pH but display their highest activity at pH 4 to 5.

cleaning supplies

The enzyme preparation is used in the context of this invention in a biocide-free cleaning agent for hard surfaces. The cleaning agent contains the enzyme preparation in amounts of 0.1 to 10.0 wt .-%, preferably 0.5 to 3 wt .-%.

In addition to the enzyme preparation, a biocide-free liquid, gelatinous or pasty aqueous cleaning agent for hard surfaces can also contain conventional cleaning agent constituents according to the invention. 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 surfactants used are preferably selected from the group of anionic surfactants, nonionic surfactants and / or amphoteric surfactants. In this case, preference is given to using anionic and / or nonionic surfactants. If used, anionic surfactants are preferably used in amounts of from 0.1 to 15% by weight, nonionic surfactants preferably in amounts of from 0.1 to 10% by weight, and amphoteric surfactants preferably in amounts of from 0.1 to 4% by weight .-% for use, in each case based on the total composition. In a less preferred embodiment, it is also possible to use cationic surfactants in amounts of up to 2% by weight, based on the total composition. Preferably, however, the cleaning agent is free of cationic surfactants due to their biocidal effect.

The anionic surfactants which can be used according to the invention include aliphatic sulfates, such as fatty alcohol sulfates, fatty alcohol ether sulfates, dialkyl ether sulfates, monoglyceride sulfates and aliphatic sulfonates, such as alkanesulfonates, .alpha.-olefinsulfonates, ether sulfonates, n-alkyl ether sulfonates, sulfonated fatty acids, ester sulfonates and lignosulfonates. Also useful in the present invention 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 (fatty acid taurides, N-acyltaurides), fatty acid sarcosinates, ether carboxylic acids, and alkyl (Ether) phosphates and α-sulfo fatty acid salts, acyl glutamates, Monoglyceriddisulfate and alkyl ethers of Glycerindisulfats, and finally their mixtures.

In the context of the present invention are fatty acids or fatty alcohols or their derivatives - unless otherwise stated - representative of 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 bis 18 carbon atoms, most preferably 12 to 16 carbon atoms, for example 12 to 14 carbon atoms. The fatty acids / alcohols or their derivatives with an even number of carbon atoms are particularly preferred because of their vegetable base as based on renewable raw materials for ecological reasons, but without the teaching of the invention is limited to them. In particular, the oxo alcohols or derivatives thereof which are obtainable, for example, by ROELEN's oxo synthesis and preferably have 7 to 19 carbon atoms, in particular 9 to 19 carbon atoms, more preferably 9 to 17 carbon atoms, most preferably 11 to 15 carbon atoms, for example 9 to 11 carbon atoms , 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 sulfonates in the form of their corresponding acid, eg. As 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 used according to the invention may also comprise soaps, ie alkali or ammonium salts of saturated or unsaturated C ₆ -C ₂₂ -fatty acids.

Preferably, the anionic surfactants are selected from the group comprising fatty alcohol sulfates in amounts of up to 5 wt .-%, alkylbenzenesulfonates in amounts of up to 7.5 wt .-% and soaps in amounts of up to 2 wt .-%, each based on the total composition, as well as mixtures thereof.

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 ₆ -C _{1 8} -Alkylalkoholpolypropylenglykol / polyethylene may 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 also be used, ie alkyl alcohol polyalkylene glycol ethers according to the above formula in which the free OH group has been etherified.

Detergents used according to the invention may contain alkyl alcohol polyglycol ethers in amounts of from 0.1 to 4% by weight, 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. While x in a given compound always has to be an integer, and above all the values x = 1 to 6, the value x for a given alkyl glycoside is 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.

Alkyl polyglycosides can be present in amounts of from 0.1 to 6% by weight, based on the total composition, in cleaners according to the invention.

As further nonionic surfactants nitrogen-containing surfactants may be included, for. As fatty acid polyhydroxy, for example glucamides, and ethoxylates of alkylamines, vicinal diols and / or carboxylic acid amides having alkyl groups having 10 to 22 carbon atoms, preferably 12 to 18 carbon atoms. The degree of ethoxylation of these compounds is generally between 1 and 20, preferably between 3 and 10. Preferred are ethanolamide derivatives of alkanoic acids having 8 to 22 carbon atoms, preferably 12 to 16 carbon atoms. Particularly useful compounds include the lauric, myristic and palmitic 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 ^- , mean in R ³ a is optionally interrupted by hetero atoms or hetero atom 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 _{1 8} 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-didecyl-ammonium chloride and their mixtures. Preferably, however, the detergent composition is free of cationic surfactants.

When choosing suitable surfactants, it must be ensured that they are compatible with the enzymes used. Preferably, the enzyme activity is lowered by not more than 50% during a typical 15 minute median exposure time, more preferably 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 less than 30%, while the activity of xylanase already reduced to less than 40% with only 0.1% SDS, so that this surfactant is not preferred for use in cleaning agents according to the invention. If this or a surfactant having a similar action is nevertheless to be used, it is preferable to ensure a spatial separation during storage, for example by encapsulating the enzymes or by using a multi-chamber bottle, so that enzymes and surfactants mix with one another only 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.

Furthermore, the agents used according to the invention may contain builders. Examples of suitable builders are alkali metal gluconates, citrates, nitrilotriacetates, carbonates and bicarbonates, in particular sodium gluconate, citrate and nitrilotriacetate, and sodium and potassium carbonate and bicarbonate, and also alkali metal and alkaline earth metal hydroxides, in particular sodium and potassium hydroxide, ammonia and amines , in particular mono- and triethanolamine, or mixtures thereof. These include the salts of glutaric acid, succinic acid, adipic acid, tartaric acid and benzene hexacarboxylic acid and phosphonates and phosphates. The agents may contain builders in amounts, based on the composition, of from 0.1 to 5% by weight.

Furthermore, the agents used 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, adipic, tartaric or malic acid, as well as other organic acids such as salicylic acid or sulfamic acid. However, the citric acid is particularly preferably used; also mixtures of several acids can be used. Acids may be present in the detergent used in the invention in amounts of up to 6 wt .-%, based on the total composition. The optionally usable bases include alkanolamines, for example mono- or diethanolamine, and ammonium or alkali metal hydroxides, especially sodium hydroxide. The cleaning agent used according to the invention may contain bases in amounts of up to 2.5% by weight, based on the total composition.

The agent used according to the invention can furthermore contain one or more thickeners for viscosity regulation. Suitable thickeners are natural and synthetic polymers and inorganic 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. B. propoxylated guar, and their mixtures, and also pectins, polyoses, locust bean gum, 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 cleaning agent used according to the invention may contain electrolyte 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 used 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 of the invention, an electrolyte salt 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 composition used in the invention, organic and / or inorganic thickeners in amounts of up to 2 wt .-%, are used, based on the total composition.

The agent used according to the invention may advantageously additionally comprise 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 used in the context of the teaching of the invention as needed in particular as a hydrotrope, viscosity regulator and / or cold stabilizer. It acts solubilizing in particular for surfactants and electrolyte as well as perfume and dye and thus contributes to their incorporation, prevents the formation of liquid-crystalline phases and has a share in the formation of clear products. The viscosity of the agent used according to the invention decreases with increasing amount of solvent. However, too much solvent can cause excessive viscosity drop. Finally, as the amount of solvent increases, the clouding and clearing point of the agent used according to the invention decreases. 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 - 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 groups per molecule, for , 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. As a solubilizer, in addition to the solvents described above, for example, alkanolamines and alkylbenzenesulfonates having 1 to 3 carbon atoms in the alkyl radical, for. As xylene or cumene sulfonate can be used. Other usable hydrotropes are z. For example, octyl sulfate or butylglucoside. The agent used in this invention may contain up to 4% by weight of these hydrotropes in amounts of, based on the total composition.

In one embodiment, the agents used according to the invention may contain abrasives. The abrasive component used may be solid water-soluble and water-insoluble, preferably inorganic compounds and mixtures thereof. These include, for example, alkali metal carbonates, alkali metal bicarbonates and alkali metal sulfates, alkali metal borates, alkali metal phosphates, silicon dioxide, crystalline or amorphous alkali metal silicates and phyllosilicates, finely crystalline sodium aluminum silicates and calcium carbonate. The advantage of water-soluble Abrasivkomponenten consists in a virtually residue-free flushability of the agent. In addition to these inorganic substances, it is also possible to use abrasives obtained from animate nature, for example ground nut shells or woods, as well as abrasion-resistant plastics, for example polyethylene beads, or even ceramic or glass beads. The cleaning agent used according to the invention may contain abrasives in amounts of up to 2% by weight, based on the total composition.

In addition to the abovementioned components, the compositions used 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 (for example 2-bromo-2-nitropropane-1,3-diol or an isothiazolinone-bromonitropropanediol preparation), perfume, dyes, pearlescing agents (for example glycol distearate) as well as opacifiers or 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.

As dyes, it is possible to use all dyes customarily used in household cleaners. Even with the fragrances all common perfumes can be used. Preference is given to fruity scents, such as citrus, pine (spruce) and mint and floral scents. Preservatives have a biocidal effect, so it is desirable to use only low concentrations, but preferably no preservatives, in detergents according to the invention.

It is advantageous if the agent contains no complexing agents; the addition of a bleaching agent to the detergent used in the invention is not required.

The pH of the agents used 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. In the case of multiphase agents, the pH of the composition means the pH of the temporary emulsion formed after shaking; in the case of compositions offered in multi-chambered bottles, the pH of the composition is that of the common intended dosage of the components stored in the various chambers components obtained solution. Thus, it is meant in each case the pH of the ready-to-use cleaning solution.

If it is a liquid agent, this preferably has a viscosity of up to 1000 mPas. By contrast, gelatinous or pasty cleaning agents can have 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 cleaning agent may contain one or more propellants (INCI propellants), usually in an amount of from 1 to 80% by weight, preferably from 1.5 to 30% by weight, in particular from 2 to 10% by weight, particularly preferably 2.5 to 8% by weight, most preferably 3 to 6% by weight. Propellants are inventively usually propellants, 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, which are generally insoluble in the liquid detergent, the operating pressure decreases with each valve actuation. Detergent-soluble or even solvent-acting liquefied gases (liquefied gases) as propellants offer the advantage of constant operating pressure and uniform distribution because the propellant vaporizes in the air, taking up more than a hundred times that volume. According to INCI suitable propellants are accordingly: butanes, carbon dioxides, dimethyl carbonates, dimethyl ether, ethanes, Hydrochlorofluorocarbon 22, hydrochlorofluorocarbon 142b, hydrofluorocarbon 152a, hydrofluorocarbon 134a, hydrofluorocarbon 227ea, isobutanes, isopentanes, nitrogen, nitrous oxides, pentanes, propanes. Chlorofluorocarbons (chlorofluorocarbons, CFCs) as propellant are, however, preferably largely and in particular completely dispensed with because of their harmful effect on the ozone shield of the atmosphere, which protects against hard UV radiation, the so-called ozone layer. Preferred blowing agents are liquefied gases. Liquefied gases are gases that can be converted from the gaseous to the liquid state at usually already low pressures and 20 ° C. In particular, under liquefied gases, however, are the hydrocarbons propane, propene, butane, butene, isobutane (2-methylpropane), isobutene (2-methylpropene), which are obtained in oil refineries as by-products from distillation and cracking of petroleum and in natural gas treatment during gasoline separation. Isobutylene) and mixtures thereof. The cleaning agent particularly preferably contains propane, butane and / or isobutane, in particular propane and butane, as one or more propellants, more preferably propane, butane and isobutane.

The cleaning agent used according to the invention can also be used in a spray dispenser. The spray dispenser is preferably a manually activated spray dispenser, in particular selected from the group comprising aerosol spray dispensers (pressurized gas containers, also known as spray can), pressure-building spray dispensers, pump spray dispensers and trigger spray dispensers, in particular pump spray dispensers and trigger spray dispensers with a transparent polyethylene or polyethylene terephthalate container. Spray dispensers are more detailed in the WO 96/04940 (Procter & Gamble) and the US patents cited therein to Sprühspendern, to which all reference is made in this regard and the contents of which are hereby incorporated into this application described. Trigger spray dispensers and pump sprayers have the advantage over compressed gas tanks that no propellant must be used.

In a further preferred embodiment, however, the agent containing the enzyme preparation is not atomized as an aerosol, since in this case possibly small amounts of the enzyme preparation can get into the respiratory tract and under certain circumstances trigger allergic reactions there. By means of suitable particles-passing attachments, nozzles, etc. (so-called "nozzle valves") on the spray dispenser, the enzyme can be added to the agent in a form immobilized on particles and dosed as a cleaning foam. When producing these compact foams, no respirable particles are formed, so that the danger of inhaling allergens is substantially eliminated.

For the hydrolysis of biofilms on hard surfaces in the household or the cleaning agent used in the invention is applied to the biofilm-coated surface and rinsed with clear water after an exposure time of 1 to 30 minutes, in case of heavy infestation even a few hours or overnight. The agent may also be blurred or triturated on the biofilm-infested surface prior to rinsing with a cloth, sponge, brush or other suitable cleaning implement on what may be e.g. B. in the presence of abrasives in the cleaning detergent cleaning performance improved. Finally, the surface is dried with a dry cloth. If the biofilm surface is very heavily attacked, the cleaning process can then be repeated.

To test the effectiveness of the agents against biofilms, appropriate test systems were set up after a detailed examination of WC biofilms. So far, unlike industrial systems, the structure of biofilms on household surfaces has been poorly understood. While Finch et al. (J. Appl. Bacteriology, 45, 357-364, 1978) found predominantly gram-positive micrococci and bacilli in and at the WC were reported by Scott et al. (J. Hygiene, 89, 279-293, 1982) found predominantly gram-negative enterobacteria, in addition isolated pseudomonads and staphylococci, in their investigation of the hygiene status of 250 households. However, both studies did not relate either thematically or by the nature of the sampling specifically to the underlying microbiological homeopathy biofilms. With another sampling system, Pitts et al. (Biofouling, 13, 19-30, 1998) detected 50 different microbial isolates in 3 households, some of which were gram-negative (these were mainly pseudomonads), others gram-positive, and some could not be characterized. Since the composition of bacterial biofilms on household surfaces has so far been little researched and the few research results did not allow any indication of any differences between biofilms on surfaces in industry and in private households, at first attempts were made to characterize WC biofilms. From the results obtained, model biofilms were then grown from several microorganisms that more accurately reflect the actual ratios on household surfaces than the biofilms previously studied most commonly from individual bacterial strains available in the laboratory. These model biofilms were then exposed to the action of various enzymes.

Characterization of WC biofilms

Sampling was done by mechanically removing the pad under the toilet rim at various toilet locations. From the biomass, the DNA was extracted directly by a standard method (DNA Tissue Kit from Qiagen, Hilden). The present 16 S-RNA gene fragments were amplified by PCR methods using two specific primers (mt12S / 1: GTG GAT CCA GGA TTA GAT ACC C; SSU2: ACT GGT ACC TTG TTA CGA CTT) and inserted into the vector pGEM-Teasy (Company Promega) cloned. The sequencing of the fragments of individual clones was carried out according to standard methods. The identification of the germs was carried out after alignment with the sequence database BLAST N2.2.1. The isolates found are shown in Table 1. Table 1: Germs from household toilets organisms Accession number Identity (bp) Identity (%) Location taxonomy 1 Nitrosomonas spec. AF272418 387/392 98 1 β-Proteobacteria 2 Aeromicrobium erythreum AF005021 471/484 97 1 Actinobacteria 3 Agrobacterium sanguineum α-Proteobacterium Porphyrobacter neustonensis AB021493 APR271042 AB033327 460/462 460/462 460/462 99 99 99 1 α-proteobacteria 4 Rhodanobacter lindanoclasticus AF039167 464/466 99 1 γ-Proteobacteria 5 Rhodanobacter lindanoclasticus AF039167 461/463 99 1 γ-Proteobacteria 6 Cellulomonas cellasea X79459 452/464 97 1 Actinobacteria 7 Unidentified bacterium wb 1 Mesorhizobium sp. USDA 4003 Rhizobium sp. CJ15 AF317774 AF282929 U50168 458/462 458/462 458/462 99 99 99 1 α-proteobacteria 8th Knoellina sinensis AJ294413 458/463 98 1 Actinobacteria 9 Uncultured bacterium GKS2-106 AJ290025 433/460 94 1 CFB 10 Uncultured sludge bacterium S6 AF234751 437/461 94 1 gram Positive 11 Uncultivated eubacterium WD2106 Uncultivated eubacterium WD2109 Uncultured eubacterium WD289 AJ292671 AJ292646 AJ292645 463/464 463/464 463/464 99 99 99 1 β-Poteobakterien 12 Dermacoccus nishinomiyaensis X87757 447/460 97 1 Actinobacteria 13 Unidentified bacterium wb1_I18 α-proteobacterium, Mena 25 / 4-1 AF317774 Y11584 460/460 460/460 100 100 1 α-proteobacteria 14 Brachybacterium tyrofermentans X91657 451/460 98 1 Actinobacteria 15 Brevundimonas sp. Brevundimonas vesicularis AJ227800 AJ227780 472/473 472/473 99 99 2 α-proteobacteria 16 Afipia genosp. 9 strain G8993 Afipia genosp. 9 strain G8990 U87780 U87779 464/464 464/464 100 100 2 α-proteobacteria 17 Bradyrhizobium japonicum strain USDA 59 Rhodopseudomonas sp. gc Afipia genosp. 4 strain G7812 AF293378 AF123086 U87770 469/479 469/479 469/479 97 97 97 2 α-proteobacteria 18 Stenotrophomonas sp. BO AF156709 466/472 98 2 γ-Proteobacteria 19 Bradyrhizobium japonicum strain USDA 59 Rhodopseudomonas sp. gc Afipia genosp. 4 strain G7812 AF293378 AF123086 U87770 468/479 468/479 468/479 97 97 97 2 α-proteobacteria 20 Xanthomonas campestris AF290420 385/387 99 2 γ-Proteobacteria Xanthomorias albinileans X95918 385/387 99 bacteria 21 Caulobacter spec. AF245033 410/421 97 2 α-proteobacteria 22 α-Proteobacterium MBIC1549 AB017110 470/490 95 2 α-proteobacteria 23 Sphingobacterium mizutae M58796 437/446 97 2 CFB 24 Sphingobacterium mizutae D14024 447/460 97 2 CFB 25 Leucobacter komagatae D45063 451/463 97 2 Actinobacteria 26 Stenotrophomonas maltophila isolate VUN 10,075 AF100733 461/461 100 2 γ-Proteobacteria 27 Pseudomonas pavonaceae D84019 463/463 100 2 γ-Proteobacteria (The next similar database entry is given in each case)

From this table it can be seen that the diversity was very large. Representatives from almost all taxonomic groups were detected at both locations. Only two isolates were found twice. The taxonomic assignment of the isolates was as follows: α-proteobacteria: 9 β-proteobacteria: 2 γ-proteobacteria: 6 CFB Group: 3 Actinobacteria (Gram-positive): 7

Structure of the test system

Based on the finding that household and toilet biofilms do not appear to consist predominantly of enterobacteria, but rather have a broad diversity in microorganisms, a biofilm test system was set up to include representatives of all taxonomically relevant groups. As far as possible, organisms were used which act as good biofilm-forming agents and can also be obtained in a defined form from strain collections. Thus, model biofilms were attracted to a mixed population consisting of: Dermacoccus nishinomiyaensis DSM No. 20448 (Actinobacterium) Bradyrhizobium japonicum DSM No. 1982 (Α-proteobacterium) Aquabacterium commune DSM No. 11901 (Β-proteobacterium) Xanthomonas campestris DSM No. 1526 (Γ-proteobacterium) (DSM: German Collection of Microorganisms and Cell Cultures)

Cultivation of the trunks:

• D. nishinomiyaensis, B. japonicum and X. campestris were incubated in 50 ml of liquid culture (10 g tryptone per liter, 5 g yeast extract, 5 g NaCl) at 30 ° C. and 150 rpm for 16 h. Inoculation with a single colony from an agar plate (per liter 10 g tryptone, 5 g yeast extract, 5 g NaCl, 10 g agarose).
• A. commune grows in 50 ml of CASO medium (Merck) (per liter 17 g peptone from casein, 3 g peptone from soy, 5 g NaCl, 2.5 g D (+) - glucose, 2.5 g di-potassium hydrogen phosphate ), which was 1:10 with DGHM water (German Society for Hygiene and Microbiology, DIN 10511 (5.3 ml of 6.71% CaCl ₂ .2H ₂ O solution, 1.52 ml of 10% MgSO ₄ .7H ₂ O Solution, 1000 ml H ₂ O _bidest )) was diluted (incubation 2 days at 28 ° C, 100 rpm). Inoculation was with a single colony of plate count agar.

The optical density (600 nm) of the cultures after 16 h was: Dermacoccus nishinomiyaensis 7.2 Bradyrhizobium japonicum 6.6 Aquabacterium commune 0.33 Xanthomonas campestris 2.5

The cultures were then combined, mixed and 50% sterile glycerol added so that the final concentration was 10% glycerol. The culture mix was portioned and frozen at -20 ° C.

The biofilm was grown in 48 well microtiter plates. Each 750 .mu.l of TBY medium was mixed with 75 .mu.l per well prepared as described above mixture of biofilm-producing cultures per well, the plates were sealed with Airpore Sheets (Qiagen) and plastic lid. The mixture was then incubated at 30 ° C. and 60 rpm for 66 h, the supernatant was carefully filtered off with suction and the plates were dried at room temperature for at least 24 h. These plates served as a substrate for the study of enzymatic hydrolysis.

Hydrolysis of biofilms

After drying, the biofilm was incubated with the enzyme-containing solution. 1 ml per well at a concentration of 2.5 mg / ml or 50 μl / ml were used for liquid enzymes. The incubation was carried out at 30 ° C and 450 rpm for 16 h either at pH 4 (62 ml 0.1 M citric acid, 38 ml 0.2 M Na ₂ HPO ₄ .2H ₂ O, 0.02% NaN ₃ ) or at pH 7.4 (1X PBS (Phosphate Buffered Saline, Gibco)) (per liter 0.21 g KH ₂ PO ₄ , 9.00 g NaCl, 0.726 g Na ₂ HPO ₄ .7H ₂ O, 0.02% NaN ₃ ). Thereafter, the supernatant was aspirated and washed once with PBS pH 7.4. After staining with a 0.01% solution of safranin O (15 minutes at room temperature), the supernatant was again aspirated. The dye was then extracted for 30 minutes at room temperature with shaking (600 rpm) in dimethyl sulfoxide; An absorbance measurement was carried out in the microtiter plate at 492 nm.

The results are shown in Table 2. Indicated are the extinction E after incubation of a biofilm with an enzyme (E _Ink ), the extinction on a reference plate without enzyme treatment (E _Ref ) and the percentage removal of the biofilm in the different enzymes. At pH 4, biofilm removals in the range of 9 to 70% were observed, at pH 7.4 11 to 62% of the biofilms were removed.

• Corolase^® PN-L (saure Protease, AB Enzymes, vormals Röhm Enzyme; E. C. 3.4.21.63) Einsatz je ml: 15 Uhb
• Dextranase 3F (ASA Enzyme; E. C. 3.2.1.11), Einsatz je ml: 1500 Units
• Cellulase aus A. niger (Sigma 01184, E. C. 3.2.1.4) Einsatz je ml: 1.25 Units
• Acid Xylanase P (AB Enzymes, E. C. 3.2.1.8) Einsatz je ml: 304 kUnits
• Econase BG 300 (β-Glucanase, AB Enzymes, E. C. 3.2.1.6.) Einsatz je ml: 15 kUnits

Tabelle 2: Entfernung von Biofilmen durch Enzyme pH 4 pH 7,4 Enzym E_Ink E_Ref Biofilmentfernung [%] E_Ink E_Ref Biofilmentfernung [%] Corolase 0,885 1,343 34,1 0,453 1,210 62,6 Dextranase 1,195 1,311 8,8 0,877 1,144 23,3 Cellulase 1,237 1,366 9,4 1,027 1,154 11,0 Glucanase 0,656 1,414 53,6 0,493 0,863 42,9 Xylanase 0,406 1,375 70,5 0,923 1,164 20,7 The enzymes used were the following: (The quantities given are per ml of incubation solution.) The units are stated in the respective manufacturer's instructions.)

• Corolase ^® PN-L (acid protease, AB Enzymes, formerly Röhm enzymes, EC 3.4.21.63) Use per ml: 15 Uhb
• Dextranase 3F (ASA enzymes, EC 3.2.1.11), use per ml: 1500 units
• Cellulase from A. niger (Sigma 01184, EC 3.2.1.4) Use per ml: 1.25 units
• Acid xylanase P (AB Enzymes, EC 3.2.1.8) Use per ml: 304 kUnits
• Econase BG 300 (β-glucanase, AB Enzymes, EC 3.2.1.6.) Use per ml: 15 kUnits

Table 2: Removal of biofilms by enzymes pH 4 pH 7.4 enzyme E _Ink E _Ref Biofilm removal [%] E _Ink E _Ref Biofilm removal [%] Corolase 0.885 1,343 34.1 0.453 1,210 62.6 dextranase 1,195 1,311 8.8 0.877 1,144 23.3 cellulase 1,237 1,366 9.4 1,027 1,154 11.0 glucanase 0.656 1.414 53.6 0.493 0.863 42.9 xylanase 0.406 1.375 70.5 0.923 1,164 20.7

Xylanase and glucanase at pH 4 as well as the corolase and again the glucanase at pH 7.4 proved to be particularly advantageous. A further increase can be achieved by a combination of different enzyme activities. In this case, on the one hand, individual enzymes can be selectively mixed with each other, which can be achieved in some cases significant performance improvements, but on the other hand, commercially available enzyme mixtures are suitable for use in the invention. By way of example, Viscozyme ^® , a liquid enzyme preparation from Aspergillus sp., Which is composed of various polysaccharidases, including arabanase, cellulase, β-glucanase, hemicellulase and xylanase, may be mentioned here.

Investigation of stability in toilet cleaner formulations

Various detergents containing enzymes for biofilm removal were prepared. (Indicated in% (w / v), exception marked) recipe E1 E2 E3 E4 E5 C _8-10 alkyl polyglucoside, DP 1.5 3.00% 3.00% 1.20% 3.50% 3.00% C _12-14 fatty alcohol sulfate Na - - 1.00% 2.50% - sodium citrate - - 1.60% 1.10% - Citric acid · 1H ₂ O 3.00% - 3.00% 4.60% - ethanol - - 0.50% - lactic acid - 3.00% - 1.60% salicylic acid - - 0.2% - - formic acid - - - - 0.35% sodium - - - - 2.04% Xanthan gum - - 0.25% 0.25% - Perfume 0.3 0.3% 0.40% 0.30% 0.30% dye 0.012% 0.012% <0.01% <0.001% - sodium hydroxide 0.83% 0.81% - - 1.20% Isobutane propane n-butane - - 4.30% 1.40% 0.30% - - liquid enzyme preparation (Corolase PN-L, 250 Uhb / g)) (in% v / v) 2.5% 2.5% 2.5% 2.5% 2.5% water ad 100% ad 100% ad 100% ad 100% ad 100% PH value 4.2 4.2 3.4 3.0 3.8

E1 and E2 were liquid toilet cleaners, E3 a toilet cleaner in aerosol form, E4 a wheel cleaner concentrate and E5 a wheel cleaner for dosing with a foam gun. All showed a good cleaning performance and in particular a good biofilm detachment.

The stability of the enzyme in the detergent formulation was determined as follows:
For control, the enzyme was added to a 100 mM Na phosphate buffer (pH 7.0) at a concentration of 2.5%. The cleaner formulations or control solution were incubated for 10 and 30 minutes at 40 ° C, respectively. 1 μl each of the samples were subsequently tested in the activity test according to Del Mar et al. (Del Mar et al., 1979, Anal. Biochem., 99, 316-320) using the synthetic substrate AAPF (Ala-Ala-Pro-Phe-nitroanilide) and determining the activity measured in μmol / ml.min. Table 3: Stability of Corolase in cleaners (in μmol / ml.min) 10 min, 40 ° C 30 min, 40 ° C phosphate buffer 607 ± 1.4 612 ± 24.2 E1 587 ± 29.9 460 ± 12.6 E2 545 ± 91.5 516 ± 76.9

Claims (16)

Use of a biocidal detergent for hard surfaces, which contains one or more surfactants and for destruction and removal of biofilms 0.1 to 10 wt .-% of an enzyme preparation of at least two enzymes, wherein the at least two enzymes to a polysaccharidase with a Protease or a polysaccharidase with a nuclease, for the destruction and removal of biofilms on household surfaces. Use according to claim 1, characterized in that the enzyme preparation is contained in amounts of from 0.5 to 3% by weight in the cleaning agent. Use according to one of claims 1 and 2, characterized in that the polysaccharidase is selected from the group β-glucanase, cellulase, xylanase, amylase, dextranase, glucosidase, galactosidase, pectinase, chitinase, lysozyme, alginate lyase. Use according to one of claims 1 to 3, characterized in that the protease is selected from the group subtilisin, thermolysin, pepsin, carboxypeptidase, acid protease. Use according to one of claims 1 to 4, characterized in that the nuclease is selected from the group DNAse, RNAse. Use according to one of claims 1 to 5, characterized in that the cleaning agent contains one or more surfactants selected from the group of anionic surfactants, nonionic surfactants and ampholytic surfactants. Use according to one of claims 1 to 6, characterized in that the cleaning agent contains one or more builders. Use according to one of claims 1 to 7, characterized in that the cleaning agent contains one or more acids and / or one or more alkalis. Use according to one of claims 1 to 8, characterized in that the cleaning agent contains one or more thickeners. Use according to one of claims 1 to 9, characterized in that the cleaning agent contains one or more solvents and / or solubilizers. Use according to one of claims 1 to 10, characterized in that the cleaning agent contains one or more abrasives. Use according to one of claims 1 to 11, characterized in that the cleaning agent contains one or more other auxiliaries and additives. Use according to claim 12, characterized in that the auxiliaries and additives are selected from the group comprising UV stabilizers, corrosion inhibitors, cleaning intensifiers, antistatic agents, perfumes, dyes, pearlescing agents, opacifiers, skin protection agents and mixtures. Use according to one of claims 1 to 13, characterized in that the cleaning agent has a pH of between 1 and 6.5. Use according to one of claims 1 to 14, characterized in that the viscosity of the liquid cleaning agent is up to 1000 mPa s (20 ° C, Brookfield LVDV II, 20 rpm, spindle no. 31). Use according to one of Claims 1 to 15, characterized in that the viscosity of the gelatinous or pasty detergent is up to 150 000 mPa s (20 ° C, Brookfield LVDV II, 20 rpm, spindle no. 31).