WO2006032477A1 - Pullulanases from psychrophilic organisms - Google Patents

Abstract

The invention relates to novel hydrolases, especially pullulanases, from psychrophilic organisms and to the use thereof.

Description

 Pullulanases from psychrophilic organisms

The present invention relates to novel hydrolases, in particular glycosidases, especially pullulanases, from psychrophilic organisms and their use.

Pullulanases are a class of enzymes able to cleave α-glycosidically linked polysaccharides, in particular α- (1, 4) or α- (1, 6) -glycosides. The prototype of such a polymer is the so-called pullulan, which is e.g. from the yeast-like fungus Pullularia pullulans (Aureobasisium pullulans). Pullulan consists of α- (1,6) -glycosidically linked maltotriose units. Pullulan degradation is mediated by a number of different enzyme specificities, which are referred to as pullulanases, neo-penululases, isopullulanases, or glucoamylases, depending on the site of the attack and the resulting end products. An overview is given by Doman-Pytka and Bardowski, (Grit Rev Microbiol, 2004, Vol. 30, 107-121).

The use of biotechnologically produced pullulanases in technical processes for starch degradation, in food technology or beer brewing is known. Since these processes usually take place at elevated temperature, usually thermostable enzymes from mesophilic, thermophilic or hyperthermophilic microorganisms are used for this purpose. In contrast, the use of only moderately thermostable or psychrophilic enzymes or of enzymes from such organisms has scarcely been investigated, in particular not for the purpose of cleaning surfaces and removing biofilms.

Biofilms are microbially induced deposits on different surfaces. The formation of biofilms is a sequential process that begins with the attachment of cells to a surface. The cells multiply and produce "extracellular polymeric substances" (EPS), which protect the cells from external influences, such as antibiotics or disinfectants EPS consists of proteins and polysaccharides In this polymer matrix, other suspended matter and ingredients (dirt, skin residues , Limescale, dead cells, etc.) from the medium surrounding the cell, ultimately leading to the formation of dirt and deposits on surfaces, whereby the dirt often becomes very solid with the surface connected is. An essential component of the EPS are polysaccharides, which can be composed very differently depending on the type of biofilm. Among them are often found α-glycosidically linked polysaccharides, eg α- (1, 4) or α- (1, 6) -glycosides containing the structural elements of the pullulan, also in combination with other building blocks, and by enzymes, the Pullulanabbau are capable of being attacked.

The use of enzymes to remove or facilitate the mechanical removal of biofilms, as well as the debris removal of surfaces, has already been described in the literature.

Thus, 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 bacteria contained therein. 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 is concerned 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.

WO 01/09295 claims enzymes having a β- (1,6) -endoglucanase activity and corresponding DNA sequences, furthermore an enzyme preparation for degrading β-1, 6-glucan-containing materials and their use. The β- (1,6) endoglucanases also serve to remove biofilms from various surfaces. However, this Spanish paper mainly deals with the degradation of the β-1, 6-glucan-rich cell wall of certain fungi. WO 96/36569 also 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 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.

International application WO 01/53010 also teaches an enzymatic process for removing biofilms from various surfaces washed with water or aqueous solutions in industrial systems and in the medical field, the biofilms being located above the surface of the water, ie solid / liquid / air. Interfaces, as well as 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.

WO 99/20239 describes the use of compositions for preventing plaque, starch hydrolyzing enzymes, e.g. an α-amylase or a pullulanase, and / or starch-modifying enzymes such as transglucosidases.

A problem of the hydrolases described in the prior art, however, is their temperature optimum. Hydrolases for technical applications are usually obtained from organisms whose temperature optimum is 37 ⁰ C or higher. Accordingly, one finds the highest activity in the range of> 30 ° C in the isolated from these organisms enzymes, the temperature optimum of the known pullulanases is even between 50 and 95 ⁰ C (Doman-Pytka and Bardowski, Critical Reviews in Microbiology, 30, 107- 121, 2004). In the removal of biofilms, for example in the household sector, the exposure temperature but at lower temperatures (15-20 ⁰ C), so that the efficiency of the enzymes used in their use is lowered. One encounters the same problem when using enzymes Cleaning temperature-sensitive laundry wants to use, which can be washed only at low temperature.

The object of the present invention was therefore to provide hydrolases which have a high activity at low temperatures, in particular α-1, 4- and / or α-1, 6-glycosidic activity, and thereby biofilm degradation and / or especially suitable for the removal of dirt from surfaces, especially at low temperatures, preferably better than the hydrolases described in the prior art.

Surprisingly, by screening for hydrolytic activity, a new psychrophilic bacterium - that is, a bacterium that grows preferentially at low temperatures - has now been discovered for the species Shewanella, which contains such an enzyme. The enzyme could be isolated and its sequence determined (SEQ ID NO: 1). It is believed that the first 23 amino acids of the polypeptide of SEQ ID NO: 1 represent a signal peptide that mediates the transport of the protein through the cell wall and is subsequently cleaved off. The bacterium was classified as Shewanella sp. 40-3 deposited with the DMSZ in Brunswick (Germany) (deposit number DSM 16509).

A first subject of the present invention is therefore the bacterium Shewanella sp. 40-3 and its use for the isolation of enzymes, in particular hydrolases, especially pullulanases, and its use for the degradation of polysaccharides, in particular for the degradation of pullulan, its use for the cleavage of α-1, 4- and / or α-1, 6-bonds, preferably α-1, 6 bonds between D-monosaccharides, in particular between D-glucose units, as well as its use for the degradation of biofilms and for the removal of soil residues from surfaces.

Another object of the present invention is a process for the isolation of hydrolases, preferably glycosylases, more preferably glycosidases, in particular glycosyl hydrolases after the classification of henrissat, especially pullulanases, from psychrophilic bacteria and / or gram-negative proteobacteria, in particular Shewanella sp., Especially from Shewanella sp. 40-3. The present invention therefore also enzymes, in particular hydrolases, preferably glycosylases, particularly preferably glycosidases, in particular glycosyl hydrolases after the classification of henrissat, especially pullulanases, from psychrophilic bacteria and / or gram-negative proteobacteria, in particular from Shewanella sp. Ago all Shewanella sp. 40-3, are available.

The method for isolating and / or identifying such enzymes may comprise in particular the following steps: a) Isolation of genomic DNA from a psychrophilic bacterium and / or gram-negative proteobacterium, in particular from Shewanella sp., Especially from Shewanella sp. 40-3, b) digesting the genomic DNA by means of suitable restriction endonucleases, c) incorporating the DNA bands, in particular those with a size between 2 and 15 kbp, preferably between 5 and 10 kbp, into a transfection vector, d) constructing a gene bank Transfection of a host, in particular E. coli, with the transfection vector, e) Screening of the gene bank for cells containing enzymes with desired activity, in particular hydrolase activity, preferably glycosylase activity, particularly preferably glycosidase activity, especially pullulanase activity , f) optionally isolating the plasmid with the clone having the desired activity, and g) optionally sequencing the isolated clone.

It is obvious to a person skilled in the art how, starting from nucleic acids according to SEQ ID NO: 1 or polypeptides according to SEQ ID NO: 2, it can obtain nucleic acids and polypeptides of similar structure and identical or similar function. In particular, enzymes with improved properties can also be obtained, for example by evolution experiments.

A further subject of the present invention is therefore a polynucleotide selected from the group consisting of: a) polynucleotide having a nucleic acid sequence according to SEQ ID NO: 1 or having a nucleic acid sequence from position 70 to position 4323 according to SEQ ID NO: 1, b) polynucleotide coding for a polypeptide having an amino acid sequence according to SEQ ID NO: 2 or for a polypeptide with an amino acid sequence from position 24 to position 1440 according to SEQ ID NO: 2, c) naturally occurring or artificially produced mutants or polymorphic forms or alleles of a polynucleotide according to (a) or (b), in particular those having up to 50 or 40, in particular up to 30 or 20, in particular up to 10, 5, 3 or 2 punk mutations or exactly one point mutation with respect to a polynucleotide according to (a) or (b), d) to a polynucleotide according to (a), (b) or (c) complementary polynucleotides, e) under stringent conditions with a polynucleotide according to (a), (b), (c) or (d) hybridizing polynucleotides, preferably under stringent conditions to be understood I incubation at 6O ⁰ C in a solution containing 0, IxSSC and 0.1% sodium dodecylsulfate (SDS), where 2OxSSC denotes a solution containing 3 M sodium chloride and 0.3 M sodium citrate (pH 7.0), f) polynucleotides having a sequence homology or Identity of at least 50%, preferably at least 60% or 70%, more preferably at least 80%, most preferably at least 90% or 95%, in particular at least 98% with respect to a polynucleotide according to (a), (b), ( c) or (d), g) polynucleotides consisting of at least 10, 15 or 20, preferably at least 25, 30 or 40, particularly preferably at least 50, 80 or 100, very particularly preferably at least 150 or 500 or 1000, in particular at least 1500 , 2000 or 2500 consecutive nucleic acids of a polynucleotide according to (a), (b), (c), (d) or (e), h) polynucleotides with deletions and / or insertions and / or inversions of up to 50 or 40, in particular up to 30, 20 or 10 nucleotides to ei polynucleotide according to (a), (b), (c), (d) or (e), i) polynucleotides comprising at least one of the polynucleotides mentioned under (a) to (h). In a further preferred embodiment, the nucleic acids according to the invention comprise one or more non-coding sequences, wherein the non-coding sequences are, for example, naturally occurring intron sequences or regulatory sequences, such as promoter or enhancer sequences, in particular those for the control of expression of hydrolases, preferably of glycosylases, more preferably glycosidases, in particular of glycosyl hydrolases after the classification of henrissat, especially pullulanases, and / or for controlling the expression of enzymes from psychrophilic bacteria and / or gram-negative proteobacteria, preferably Shewanella, especially Shewanella sp. 40-3, act.

The nucleic acids according to the invention are preferably ribonucleic acids (RNAs) or deoxyribonucleic acids (DNAs), the nucleic acids preferably being present as double-stranded nucleic acids. The nucleic acids according to the invention are preferably nucleic acids which code for a protein with hydrolase activity or for parts thereof, wherein the parts are in particular domains or epitopes which can be used, for example, for antibody production. According to the invention, domains are in particular also those parts of the enzyme which do not themselves have to be directly involved in the hydrolysis, but can also exert or participate in other partial or auxiliary functions within the enzyme, whereby the partial or auxiliary functions for example, the binding of a substrate, intermediate or end product, a carrier function, the mediation of a regulatory influence on the hydrolytic activity or, further, about the enzyme anchoring can act. The protein with hydrolase activity is preferably a glycosylase, more preferably glycosidase (EC3.2.1), in particular glycosylhydrolase according to the classification of henrissat given below, especially pullulanase (EC3.2.1.41), and / or an enzyme which is capable of, α-1, 4 and / or α-1, 6 bonds, more preferably α-1, 6 bonds, between D-monosaccharides, in particular between D-glucose units to split. By cleavage according to the invention is meant above all hydrolytic cleavage, ie cleavage to release the respective sugar residues. Most preferably, the hydrolase or pullulanase is a hydrolase with (β / α) δ-folding ((β / α) ₈ -Barrel structure) and / or a hydrolase comprising such a structural element.

In a preferred embodiment, a polypeptide of the invention be temperature optimum, especially in relation to the biofilm degradation, at a temperature between 4 and 4O ⁰ C, more preferably between 10 and 35 ⁰ C, especially 15 to 30 ⁰ C. The pH Optimum of a polypeptide according to the invention is preferably between pH 5 and pH 9, more preferably between pH 6 and pH 8, especially between pH 6.5 and pH 7.5.

The nucleic acids according to the invention are preferably nucleic acids which are suitable for a pullulanase, particularly preferably for a microbial pullulanase, especially a bacterial pullulanase, in particular for a pullulanase from a psychrophilic bacterium and / or a gram-negative proteobacterium, preferably from Shewanella sp., especially from Shewanella sp. 40-3 and / or for a pullulanase according to SEQ ID no. 2 encode.

The present invention furthermore relates to the use of the nucleic acids according to the invention, on the one hand for the production or isolation of nucleic acids according to the invention, on the other hand for the preparation or isolation of novel nucleic acids homologous to the nucleic acids according to the invention, in particular those with structural and similar properties which are similar with respect to the nucleic acids according to the invention , similar and / or improved functional properties, wherein among functional properties in particular hydrolase activity, preferably glycosylase activity, particularly preferably glycosidase activity, especially pullulanase activity and improved functional properties, for example, higher specificity and / or higher conversion and / or higher stability of the enzyme at the desired reaction conditions, in particular with respect to the pH, the temperature and with respect to the optionally added detergents and proteases, to ve stand up. The nucleic acids according to the invention can thus be used, for example, as probes for the identification and / or isolation of homologous nucleic acids from an artificial, cDNA or genomic gene bank, in this case preferably for the identification of nucleic acids which are suitable for hydrolases, in particular glycosylases, particularly preferably glycosidases, in particular glycosylhydrolases according to US Pat Classification of Henrissat, especially pullulanases and / or encode parts thereof, or as antisense nucleic acids or as a primer in the polymerase chain reaction (PCR), in particular for the amplification of nucleic acids comprising nucleic acids encoding hydrolases, preferably glycosylases, more preferably glycosidases ago glycosyl hydrolases after the classification of Henrissat, in particular for pullulanases or for parts thereof.

However, the nucleic acids according to the invention or homologous to the nucleic acids according to the invention can also be obtained by random mutagenesis or targeted mutagenesis, in a manner known to the person skilled in the art.

After incorporation into an expression vector, the nucleic acids according to the invention can be used, for example, for the targeted production of individual domains or epitopes of the protein according to the invention or for the preparation of fusion proteins which comprise the polypeptides according to the invention.

The present invention therefore also provides a process for obtaining a nucleic acid which codes for a hydrolase, preferably glycosylase, particularly preferably glycosidase, in particular pullulanases or parts thereof, comprising the following steps: a) a nucleic acid library is contacted with a nucleic acid according to the invention , b) a nucleic acid which hybridizes with the nucleic acid according to the invention according to (a) is identified, c) the nucleic acid identified in step (b) is sequenced. A further subject of the present invention is therefore likewise a process for isolating a nucleic acid coding for a hydrolase, preferably glycosylase, more preferably glycosidase, in particular pullulanase or for parts thereof, comprising the following steps: a) primers are prepared starting from a nucleic acid according to the invention, b) the primers according to (a) are used to amplify in the PCR nucleic acids, especially cDNAs, of unknown nucleic acid sequence, c) the nucleic acids obtained according to (b) by amplification are sequenced.

A further subject of the present invention is therefore also a process for isolating a nucleic acid coding for a hydrolase, preferably glycosylase, more preferably glycosidase, in particular pullulanase or for parts thereof, comprising the following steps: a) nucleic acids of a library are incorporated into suitable vectors and these are incorporated in b) The host organisms obtained according to (a) are plated on a solid support, preferably an agar plate, and incubated on the solid Carrier is assayed, for example, a color reaction involving the reaction of a hydrolase substrate, c) hydrolase-positive clones are identified, for example, by visual observation, by automated image analysis or by photometric measurement, and purified, d) the nucleic acidsthe clones identified in step (c) are sequenced.

Another object of the present invention is therefore likewise a process for isolating a nucleic acid coding for a hydrolase, preferably glycosylase, particularly preferably glycosidase, in particular pullulanase or for parts thereof, comprising the following steps: a) a nucleic acid according to the invention is subjected to mutagenesis experiments, b) the resulting mutants are incorporated into a suitable vector and expressed in a suitable host organism, c) the expression products are examined for hydrolytic activity, in particular pullulanase activity, d) the nucleic acids whose expression products in step (c) show the desired hydrolytic activity are sequenced.

The mutagenesis experiments can be carried out, for example, using the polymerase chain reaction (PCR). The experiments can be carried out in this case, for example, in particular when using a Taq polymerase, so that reaction parameters such as the Mg 2+ concentration, the pH, the reaction temperature or the substrate concentrations are varied, or it can be used for error-prone PCR Techniques, for example, based on the addition of Mn2 + or on the addition of unequal nucleotide concentrations.

The nucleic acid library to be used according to the invention may be, for example, a cDNA library, a genomic library or an artificial library. It is preferably a microbial, particularly preferably a bacterial library, in particular of psychrophilic bacteria, in particular of Shewanella, in particular of Shewanella sp. 40-3.

The invention further provides a nucleic acid obtainable by one of the aforementioned methods.

Another object of the present invention is a process for the preparation of a nucleic acid according to the invention, characterized in that the nucleic acid is chemically synthesized. The nucleic acids according to the invention can be used, for example, chemically based on the amino acid sequences shown in SEQ ID NO. 1 or based on the genetic code shown in SEQ ID NO: 2 synthesized on the basis of the genetic code, for example by the Phosphotriester method or in another manner known in the art. The present invention furthermore relates to vectors, in particular cloning and / or expression vectors, comprising one of the abovementioned nucleic acids according to the invention. The vector may be, for example, a prokaryotic or eukaryotic vector. The prokaryotic vectors for incorporation of the nucleic acids according to the invention are, for example, the plasmid pBK-CMV, the plasmid pBR322, plasmids of the pUC series, derivatives of pUB 110 or other plasmids suitable for expression in gram-negative or Gram-positive bacteria are suitable.

However, the expression vectors for expression in E. coli can also be, for example, other commercially available vectors, such as the T7 expression vector pGM10 or pGEX-4T-1 GST (Pharmacia Biotech), which are suitable for an N-terminal metabolic vector. Ala-His6 tag, which allows purification of the expressed protein via a Ni2 + -NTA column. As eukaryotic expression vectors for expression in Saccharomyces cerevisiae, e.g. the vectors p426Met25 or p426GAL1, for expression in insect cells e.g. Baculovirus vectors as disclosed in EP-B1-0127839 or EP-B1-0549721, and for expression in mammalian cells e.g. SV40 vectors.

In one embodiment, the nucleic acids according to the invention can be incorporated with flanking nucleic acids into a vector such that the expression of the vector encodes the polypeptides encoded by the nucleic acids according to the invention as fusion proteins or carries a tag, a tagging amino acid sequence, for example purification and / or detection can facilitate the polypeptides. The tag may be, for example, the strep, flag, myc or his tag.

The expression vectors preferably contain the requlatory sequences suitable for the host cell, in this case preferably the lac or the tac promoter for expression in E. coli, the ADH-2, GAL1 or AOX promoter for expression in yeasts, the baculovirus Polyhedrin promoter for the

Expression in insect cells or the SV40 early promoter or an LTR promoter for expression in mammalian cells. A further subject of the present invention is a host cell comprising a vector according to the invention, wherein the host cell is preferably Shewanella, in particular Shewanella sp. 40-3, or E. coli, in particular E. coli strain BL21 (DE3), or Bacillus, in particular Bacillus licheniformis, Bacillus amyloliquefaciens, Bacillus subtilis or Bacillus alcalophilus.

After incorporation into a suitable vector, the nucleic acids according to the invention are preferably introduced into the host cells by the methods of transfection, transformation or electroporation known to the person skilled in the art.

Another object of the present invention is a polypeptide selected from the group consisting of: a) polypeptide having an amino acid sequence according to SEQ ID NO: 2 or from position 24 to position 1440 according to SEQ ID NO: 2, b) naturally occurring mutants, polymorphic forms or Alleles of a polypeptide according to (a), especially those having up to 50, 20, 15 or 10 point mutations, in particular up to 5, 3 or 2 point mutations or exactly one point mutation, with respect to a polypeptide according to (a), c ) Polypeptides having a sequence homology or identity of at least 50 or 55%, preferably at least 60, 65 or 70%, very particularly preferably at least 75, 80, 85 or 90%, in particular at least 91, 92, 93, 94, 95, 96, 97, 98 or 99% with respect to a polypeptide according to (a) or (b), d) polypeptides which are encoded by the aforementioned nucleic acids according to the invention, e) polypeptides consisting of at least 5 or 6, preferably min at least 8, 10, 12 or 15, more preferably at least 20, 25 or 50, especially at least 100, 200 or 500, especially at least 800, 1000 or 1200 consecutive amino acids of a polypeptide according to (a) or (b), f) polypeptides with insertions and / or deletions and / or inversions of up to 60, 50 or 40, in particular of up to 30, 20 or 10, especially of one, two or three amino acids in relation to a polypeptide according to (a), (b ) or (c), g) polypeptides comprising at least one of the polypeptides mentioned under (a) to (f). These are preferably polypeptides which catalyze a hydrolysis reaction, in particular the hydrolysis of α-1, 4- and / or α-1, 6 bonds, more preferably of α-1, 6 bonds, between D and Monosaccharides, in particular between D-glucose units, or are parts, in particular epitopes or domains, of such polypeptides, which according to the invention include domains according to the invention, in particular those parts of the enzyme, which need not be directly involved in the hydrolysis themselves but may also perform or participate in other partial or auxiliary functions within the enzyme, the partial or auxiliary functions being, for example, the binding of a substrate, an intermediate or end product, a carrier function, mediation a regulating influence on the hydrolytic activity or about the enzyme anchoring can act. The epitopes and / or domains can be present both isolated and as part of a chimeric protein and / or as part of a fusion protein.

The protein with hydrolase activity is preferably a glycolase, more preferably glycosidase (EC3.2.1), especially pullulanase (EC3.2.1.41) and / or it is a hydrolase with (β / α) s-folding ((β / α) s-barrel structure) and / or it is a hydrolase which belongs to the family of glycosyl hydrolases according to the classification of henrissat given below or is homologous to these enzymes and / or it is an enzyme that is suitable for biofilm degradation.

The classification of glycosyl hydrolases is based on the classification of Henrissat (Henrissat B., Bairoch A. Updating the Sequence-based Classification of glycosyl hydrolases, Biochem J. 316: 695-696 (1996)). The current version of the mapping is in the CAZY database (Coutinho, PM & Henrissat, B. (1999) Carbohydrate-Active Enzymes server at URL: http://afmb.cnrs-mrs.fr/~cazy/CAZY/index. html). Thereafter, pullulanases and related enzymes are found in the glycosyl hydrolase (GHF) family 13. In addition to the common (β / α) s barrel structure, these enzymes are characterized by the ability to hydrolyze α-linked glycosidic bonds. In addition to pullulanases (EC 3.2.1.41) and neo-foldulanases (EC 3.2.1.135), there are also amylases (EC 3.2.1.1), cyclomaltodextrin glucanotransferase (EC 2.4.1.19), Cyclomaltodextrinase (EC 3.2.1.54), trehalose-6-phosphate hydrolase (EC 3.2.1.93), oligoglucosidase (EC 3.2.1.10), maltogenic amylase (EC 3.2.1.133), alpha-glucosidase (EC 3.2.1.20), maltotetraose forming amylase (EC 3.2.1.60), isoamylase (EC 3.2.1.68), glucodextranase (EC 3.2.1.70), maltohexaose-forming amylase (EC 3.2.1.98), branching enzymes (EC 2.4.1.18), trehalose synthase (EC 5.4 .99.16), 4-glucanotransferase (EC 2.4.1.25), maltopentaose-forming amylase (EC 3.2.1.-), amylosucrase (EC 2.4.1.4), sucrose phosphorylase (EC 2.4.1.7), malto-oligosyl-trehalose trehalohydrolase (EC 2.4.1.141) and isomaltulose synthase (EC 5.4.99.11).

The polypeptides of the invention preferably comprise four highly conserved regions typical of pullulan degrading enzymes. These form the reaction center and are involved in substrate binding. These conserved regions are characterized, for example, by Doman-Pytka and Bardowski (Critical Reviews in Microbiology, 30, 107-121, 2004).

The polypeptides of the invention may also be post-translationally and / or chemically modified.

The compounds which can be cleaved by the polypeptides according to the invention are, in particular, compounds which contain D monosaccharides, in particular D-glucose units, which have α-1, 4- and / or α-1, 6 Bonds are linked together. These are preferably pullulan, but they may also be other oligo- or polysaccharides containing such bonds, for example amylopectin, amylose, dextrins and glycogen.

The present invention therefore also provides a process for the cleavage of the aforementioned compounds, characterized in that these compounds are incubated with at least one polypeptide of the invention. Another object of the invention is therefore also the use of a polypeptide according to the invention for the hydrolytic cleavage of α-1, 4- and / or α-1, 6 bonds, more preferably of α-1, 6 bonds, between D-monosaccharides, in particular between D-glucose units, and the use of a polypeptide according to the invention for the degradation of biofilms and / or for the prevention of the formation of biofilms and the removal of soil residues from surfaces.

In a particularly preferred embodiment, the hydrolases according to the invention are microbial, in particular bacterial, hydrolases. Very particular preference is given to a hydrolase, in particular pullulanase, from a psychrophilic bacterium and / or from a gram-negative proteobacterium, in particular from Shewanella sp., More preferably from Shewanella sp. 40-3, especially the pullulanase with a sequence according to SEQ ID NO: 2.

In a further preferred embodiment, the polypeptide according to the invention is a water-soluble pullulanase. Depending on the method of preparation, in particular depending on the host cells and vectors used and depending on optionally used signal sequences, the polypeptide can be produced in the cytosol, in the periplasm or in the form bound to the cytosolic or outer membrane or into the surrounding Medium are discharged, the latter being preferred.

The present invention furthermore relates to mixtures and preparations, in particular bacterial preparations, which contain nucleic acids and / or polypeptides according to the invention. The mixtures or preparations can in this case be prepared in a manner known to the person skilled in the art. The polypeptides of the invention may further be used, for example, as epitopes for the production of monoclonal or polyclonal antibodies by coupling to a carrier, for example bovine serum albumin, followed by a mammal, preferably a mouse, rabbit or rabbit the epitope, preferably using adjuvants. Preferred for this are polypeptides with a length of 5-12, in particular 8, amino acids. Polypeptides longer than 60 amino acids, especially greater than 75 amino acids, can also be used without carriers to produce antibodies. The resulting antibodies may then optionally be isolated, and optionally antibody fragments, for example Fab or scFv fragments, can be prepared starting from the antibodies or the nucleic acids coding for them.

Alternatively, peptides which bind to a polypeptide according to the invention can also be obtained by a method known to those skilled in the art, such as phage display, yeast display, bacterial display or the so-called fusagene technology, in which the nucleic acid and the polypeptide encoded by it are Puromycin are covalently linked together.

The antisera, antibodies and antibody fragments obtainable by immunization with the polypeptides according to the invention and the peptides obtainable by one of the mentioned in vitro methods are suitable, for example, for the investigation of gene expression libraries, to proteins homologous to the polypeptides according to the invention, in particular those with hydrolytic activity, especially esterolytic and / or lipolytic activity.

The present invention therefore also provides antisera, antibodies and antibody fragments against a polypeptide according to the invention and other peptides which bind to a peptide according to the invention, in particular obtainable by one of the abovementioned methods.

Antiserum, antibody and antibody fragment as well as other peptides binding to a peptide according to the invention are referred to below as "antibodies" for the sake of simplicity. The present invention further provides a process for producing a polypeptide according to the invention, characterized in that a nucleic acid according to the invention in a suitable host cell, more preferably in E. coli or Shewanella, in particular in Shewanella sp. 40-3, or expressed in Bacillus, in particular in Bacillus iformis, Bacillus amyloliquefaciens, Bacillus subtilis or Bacillus alcalophilus. Subsequently, if necessary, a protein purification can take place.

Cultivation of E. coli is preferably carried out between 20 and 37 ° C, more preferably at about 30 ⁰ C or at about 37 ° C. Harvest and digestion of the cells are carried out by methods known to those skilled in the art. The digestion can be done for example by French Press, ultrasound, ball mill, but more preferably by using a Sonifikators. Alternatively, the cells may also be chemically permeabilized, for example by EDTA or polymyxin B. The optionally carried out purification of the protein is preferably carried out by chromatography. The chromatographic purification of the protein may include, for example, cation and / or anion exchange chromatography, hydrophobic interaction chromatography and / or gel filtration.

In particular, the short-chain polypeptides according to the invention can also be synthesized by means of classical peptide synthesis (Merrifield technique).

According to the invention, one or more nucleotides of the nucleic acid or one or more amino acids of the polypeptides or antibodies can also be modified. The modification may, for example, be a radioactive, fluorogenic or chromogenic group or a post-translational modification.

The present invention furthermore relates to aqueous preparations (enzyme preparations) which contain the polypeptides according to the invention in dissolved form. The enzyme preparations according to the invention are described in more detail below.

Another object of the present invention is any type of composition containing a polypeptide of the invention. It can vary depending on Intended use of a solid mixture, for example, a powder with freeze-dried or encapsulated polypeptides, or to act a gelatinous, pasty or liquid composition. As an application form in particular extrudates, granules, tablets and pouches in question.

The encapsulated form lends itself to protecting the enzymes or other ingredients from other ingredients, such as bleaches, or to allow for controlled release. Depending on the size of these capsules, a distinction is made to MiIIi-, micro- and nanocapsules, with microcapsules for enzymes being particularly preferred.

In the case of solid agents, the proteins can be used, for example, in dried, granulated and / or encapsulated form. They may be added separately, ie as a separate phase, or with other ingredients together in the same phase, with or without compaction. If microencapsulated enzymes are to be processed in solid form, the water can be removed by methods known from the prior art from the aqueous solutions resulting from the workup, such as spray drying, centrifuging or by solubilization. The particles obtained in this way usually have a particle size between 50 and 200 microns.

The enzymes, and also the protein according to the invention, can be added to liquid, gelatinous or pasty agents according to the invention starting from a protein recovery and preparation carried out in concentrated aqueous or non-aqueous solution, suspension or emulsion, but also in gel form or encapsulated or as dried Powder. Such detergents or cleaners according to the invention are generally prepared by simple mixing of the ingredients which can be added in bulk or as a solution in an automatic mixer.

With regard to possible fields of use of the polypeptides according to the invention, reference is made to the manual "Industrial Enzymes and their Applications" by H. UhMg, Wiley-Verlag, New York, 1998. Possible industrial applications of the polypeptides according to the invention and / or of the enzyme preparations according to the invention are, for example, the production or selective enrichment of flavor components in the food industry, fine chemicals, in particular sugar, in the chemical industry or therapeutically or diagnostically usable substances in the pharmaceutical industry, and further processing from starch to amylose, glucose syrup, maltose syrup, dextrins, maltose or high purity glucose or fructose, the degradation of pullulan to panose, which can be used to prevent tooth decay (Kuriki et al., J. Bacteriol 170, 1554, 1988) , the treatment of foods and the cleavage of cereals and cereal products in the brewing industry, in particular for the production of "light" beer The use can also be used in combination with other hydrolases, in particular glycosidases, for example in combination with α-amylases or Glucoamyla sen, done.

The present invention furthermore relates to the use of the polypeptides according to the invention and / or the enzyme preparations according to the invention, in particular in sterilization, disinfection, washing, dishwashing and cleaning agents, in particular in cleaners for removing biofilms and / or for removing organic Dirt from hard surfaces. The use may in particular in the household or in the field of pharmaceutical, food, brewery, medical technology, paint, wood, textile, cosmetics, leather, tobacco, fur, rope, paper , Pulp, plastics, fuel, oil, rubber or machinery industry or in dairies. The polypeptides according to the invention can be used in particular for the removal of biofilms from household and / or sanitary articles as well as in pulp and paper mills, in circulation cooling towers and in other systems which lead flowing and / or circulating water. In addition to the removal of biofilms, the polypeptides according to the invention are, for example, also suitable for use in detergents for cleaning textiles, in particular for stain removal.

The present invention therefore furthermore also includes sterilization, disinfecting, washing, dishwashing and cleaning agents which contain polypeptides according to the invention and / or enzyme preparations according to the invention. In which Means can basically be a means for any type of surface. Thus, the surface may be a biotic or abiotic, artificially synthesized or natural, soft or hard surface. The surface may be, for example, a textile, ceramic, metal and / or plastic surface. The article may be, for example, laundry, dishes, sanitary equipment, floor coverings, shoes, leather, rubber articles, ship hulls, prostheses, teeth, dentures or catheters.

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, the cleaner is one for cleaning hard surfaces, such as floors, tiles, dishes, tiles, plastics, and other hard surfaces in the home, industrial, public sanitation, swimming pools, Saunas, sports facilities or in doctor or massage practices.

Medically relevant biofilms that can be removed with poypeptides according to the invention are, for example, biofilms from chronic lung infections, e.g. in cystic fibrosis patients. Furthermore, it may be plaque or biofilms that must be removed from contact lenses, implants or medical devices, instruments or equipment such as catheters or endoscopes.

The present invention therefore also relates to the use of the polypeptides according to the invention and / or the enzyme preparations according to the invention in cosmetic and / or pharmaceutical products as well as cosmetic and / or pharmaceutical products which contain the polypeptides and / or enzyme preparations according to the invention as well as the use of the polypeptides according to the invention and / or enzyme preparations for the production of cosmetic and / or pharmaceutical products. For example, the product may be a dental plaque removal product or a biofilm removal product caused by or associated with chronic lung infections, such as in the art Cystic fibrosis case, act. The present invention, therefore, especially oral, dental and dental prosthesis care agents containing the polypeptides of the invention.

The abovementioned agents and products according to the invention may comprise further components as are known to the person skilled in the art. Here, the sterilization, disinfection, washing, dishwashing, textile treatment and cleaning agents, depending on the intended use, for example, one or more components selected from the group consisting of surfactants, builders, acids, alkaline substances, hydrotropes, solvents, thickeners, bleaching agents, Dyes, perfumes, corrosion inhibitors, sequestering agents, electrolytes, optical brighteners, grayness inhibitors, silver corrosion inhibitors, color transfer inhibitors, foam inhibitors, abrasives, UV absorbers and skin protection agents. To stabilize the polypeptide, enzyme stabilizers may also be added. 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, a gel, a paste or a powder.

Polypeptides or enzyme preparations according to the invention are preferably present in agents and / or compositions according to the invention in an amount of up to 20% by weight, more preferably in an amount of 0.01-10% by weight.

enzyme preparation

The enzyme preparation according to the invention is a system, in particular an aqueous concentrate, containing one or more of the polypeptides according to the invention. Optionally, other enzymes, in particular selected from polysaccharidases, proteases and nucleases, may also be present in the enzyme preparation.

The polysaccharidase may in particular be a β-glucanase, cellulase, hemicellulase, arabinase, xylanase, amylase, dextranase, glucosidase, galactosidase, pectinase, chitinase, lysozyme, alginate lyase or any mixtures thereof. In particular, the protease may be subtilisin, thermolysin, pepsin, a carboxypeptidase, an acidic protease, a Serine proteinase or mixtures thereof. The nucleases may be any DNAse or RNAse.

The enzymes which can additionally be used in the enzyme preparations 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.

Regarding the use of enzymes, it should be noted in principle, regardless of whether they are polypeptides according to the invention or other enzymes, that they could be attacked in liquid agents by components contained in the cleaner, such as surfactants or organic acids, or else by non-specifically acting proteases , which would reduce the effectiveness of the enzymes. 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 purification 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 cleaning or washing, by sudden change in osmotic conditions, for example when diluting with water or by cutting the capsules during dosing out of the storage bottle, to be opened 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 are also each immobilized on a solid support, which is suspended in the detergent and can additionally serve as 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 or enzyme groups 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. Thus it is 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 this with the enzyme storage solution, their pH causes an optimum of enzyme activity. Components for Detergents and Cleaning Agents According to the Invention The surfactants which can be used according to the invention are selected from the groups of anionic, nonionic, amphoteric and cationic surfactants and mixtures thereof, the anionic, nonionic and amphoteric surfactants and mixtures thereof being preferred and the anionic and nonionic surfactants and mixtures of which are particularly preferred.

The nonionic surfactants used are preferably alkoxylated, advantageously ethoxylated, in particular primary, alcohols having preferably 8 to 18 carbon atoms and on average 1 to 12 moles of ethylene oxide (EO) per mole of alcohol, in which the alcohol radical can be linear or preferably methyl-branched in the 2-position , or may contain linear and methyl-branched radicals in the mixture, as they are usually present in Oxoalkoholresten. In particular, however, alcohol ethoxylates with linear radicals of alcohols of natural origin having 12 to 18 carbon atoms, for example from coconut, palm, tallow or oleyl alcohol, and on average 2 to 8 EO per mole of alcohol are preferred. Preferred ethoxylated alcohols include, for example, Ci ₂ -i ₄ alcohols containing 3 EO or 4 EO, n-alcohol with 7 EO, C _3- I ₅ alcohols containing 3 EO, 5 EO, 7 EO or 8 EO, C ₁₂ -i ₈ -alcohols with 3 EO, 5 EO or 7 EO and mixtures of these, such as mixtures of Ci _2- i ₄ -alcohol with 3 EO and Ci _2- i ₈ -alcohol with 5 EO. The degrees of ethoxylation given represent statistical means which, for a particular product, may be an integer or a fractional number. Preferred alcohol ethoxylates have a narrow homolog distribution (narrow rank ethoxylates, NRE). In addition to these nonionic surfactants, fatty alcohols with more than 12 EO can also be used. Examples of these are tallow fatty alcohol with 14 EO, 25 EO, 30 EO or 40 EO.

Another class of preferred nonionic surfactants used either as the sole nonionic surfactant or in combination with other nonionic surfactants are alkoxylated, preferably ethoxylated or ethoxylated and propoxylated fatty acid alkyl esters, preferably having from 1 to 4 carbon atoms in the alkyl chain, especially fatty acid methyl esters.

Another class of nonionic surfactants that can be used to advantage are the alkyl polyglycosides (APG). Usable Alkypolyglycosides satisfy the general formula RO (G) _Z in which R is a linear or branched, especially in the 2-position methyl-branched, saturated or unsaturated, aliphatic radical having 8 to 22, preferably 12 to 18 carbon atoms and G is the symbol, which represents a glycose unit having 5 or 6 C atoms, preferably glucose. The degree of glycosylation z is between 1, 0 and 4.0, preferably between 1, 0 and 2.0 and in particular between 1, 1 and 1, 4. Preference is given to using linear alkyl polyglucosides, that is to say alkyl polyglycosides in which the polyglycosyl radical is a glucose radical and the alkyl radical is an n-alkyl radical.

Nonionic surfactants of the amine oxide type, for example N-cocoalkyl-N, N-dimethylamine oxide and N-tallowalkyl-N, N-dihydroxyethylamine oxide, and the fatty acid alkanolamides may also be suitable. The proportion of these nonionic surfactants is preferably not higher than that of the ethoxylated fatty alcohols, especially not more than half of them.

Further suitable surfactants are polyhydroxy fatty acid amides of the formula (II)

R ¹

I

R-CO-N- [Z] (II)

in the RCO for an aliphatic acyl radical having 6 to 22 carbon atoms, R ^{1 is} hydrogen, an alkyl or hydroxyalkyl radical having 1 to 4 carbon atoms and [Z] for a linear or branched polyhydroxyalkyl radical having 3 to 10 carbon atoms and 3 to 10 hydroxyl groups stands. The polyhydroxy fatty acid amides are known substances which can usually be obtained by reductive amination of a reducing sugar with ammonia, an alkylamine or an alkanolamine and subsequent acylation with a fatty acid, a fatty acid alkyl ester or a fatty acid chloride.

The group of polyhydroxy fatty acid amides also includes compounds of the formula

(III).

I

R-CO-N- [Z] (III) in the R is a linear or branched alkyl or alkenyl radical having 7 to 12 carbon atoms, R ^{1 is} a linear, branched or cyclic alkyl radical or an aryl radical having 2 to 8 carbon atoms and R ^{2 is} a linear, branched or cyclic alkyl radical or an aryl radical is or an oxyalkyl group having 1 to 8 carbon atoms, Ci _-4 alkyl or phenyl groups being preferred, and [Z] is a linear polyhydroxyalkyl residue, whose alkyl chain is substituted with at least two hydroxyl groups, or alkoxylated, preferably ethoxylated or propoxylated derivatives this rest.

[Z] is preferably obtained by reductive amination of a reducing sugar, for example glucose, fructose, maltose, lactose, galactose, mannose or xylose. The N-alkoxy- or N-aryloxy-substituted compounds can be converted into the desired polyhydroxy fatty acid amides, for example, by reaction with fatty acid methyl esters in the presence of an alkoxide as catalyst.

As anionic surfactants, for example, those of the sulfonate type and sulfates are used. The surfactants of the sulfonate type are preferably C ₉ . ₁₃ - alkyl benzene sulfonates, olefin sulfonates, ie mixtures of alkene and hydroxyalkane sulfonates, and the disulfonates obtained, for example, from C ₁₂ - receives monoolefins with terminal or internal double bond by sulfonation with gas¬ shaped sulfur trioxide and subsequent alkaline or acidic hydrolysis of the sulfonation - ₁₈ , considering. Also suitable are alkanesulfonates, from C ₂ - ₁₈ are obtained alkanes, for example by sulfochlorination or sulfoxidation and an¬ subsequent hydrolysis or neutralization. Likewise suitable are the esters of .alpha.-sulfo fatty acids (ester sulfonates), for example the .alpha.-sulfonated methyl esters of hydrogenated coconut, palm kernel or tallow fatty acids.

Further suitable anionic surfactants are sulfated fatty acid glycerol esters. Fatty acid glycerol esters are to be understood as meaning the mono-, di- and triesters and mixtures thereof, as obtained in the preparation by esterification of a monoglycerol with 1 to 3 moles of fatty acid or in the transesterification of triglycerides with 0.3 to 2 moles of glycerol. Preferred sulfated fatty acid glycerol esters are the Sulfierprodukte of saturated fatty acids having 6 to 22 carbon atoms, spielsweise of caproic acid, caprylic acid, capric acid, myristic acid, lauric acid, palmitic acid, stearic acid or behenic acid.

Alk (en) yl sulfates are the alkali and especially the sodium salts of the Schwefelsäurehalbester _Ci2 -Ci8 fatty alcohols, for example, from coconut fatty alcohol, tallow fatty alcohol, lauryl, myristyl, cetyl or stearyl alcohol, or C ₁₀ -C 2 ₀ - oxo alcohols, and those half-esters of secondary alcohols of these chain lengths are preferred. Also preferred are alk (en) ylsulfates of said chain length, which contain a synthetic, produced on a petrochemical basis straight-chain alkyl radical, which have an analogous degradation behavior as the adequate compounds based on oleochemical raw materials. Of washing technology interest, the Ci ₂ -Ci ₆ alkyl sulfates and C ₁₂ -Ci ₅ alkyl sulfates and C ₁₄ -C ₁₅ - alkyl sulfates are preferred. 2,3-alkyl sulfates are also suitable anionic surfactants.

Also, the sulfuric monoesters of ethoxylated with 1 to 6 moles of ethylene oxide straight or branched Cz ^ i alcohols, such as 2-methyl-branched Cg-n-alcohols having an average of 3.5 moles of ethylene oxide (EO) or C _12-18 fatty alcohols 1 to 4 EO, are suitable. Due to their high foaming behavior, they are only used in detergents in relatively small amounts, for example in amounts of up to 5% by weight, usually from 1 to 5% by weight.

Further suitable anionic surfactants are also the salts of alkylsulfosuccinic acid, which are also referred to as sulfosuccinates or as sulfosuccinic acid esters and the monoesters and / or diesters of sulfosuccinic acid with alcohols, preferably fatty alcohols and in particular ethoxylated fatty alcohols. Preferred sulfosuccinates contain Cs-iβ fatty alcohol residues or mixtures of these. Particularly preferred sulfosuccinates contain a fatty alcohol radical which is derived from ethoxylated fatty alcohols, which in themselves constitute nonionic surfactants (description see above). Sulfosuccinates whose fatty alcohol residues are derived from ethoxylated fatty alcohols with a narrow homolog distribution are again particularly preferred. Likewise, it is also possible to use alk (en) ylsuccinic acid having preferably 8 to 18 carbon atoms in the alk (en) yl chain or salts thereof. As further anionic surfactants are particularly soaps into consideration. Suitable are saturated fatty acid soaps, such as the salts of lauric acid, myristic acid, palmitic acid, stearic acid, hydrogenated erucic acid and behenic acid and, in particular, soap mixtures derived from natural fatty acids, for example coconut, palm kernel or tallow fatty acids.

The anionic surfactants, including the soaps, may be present in the form of their sodium, potassium or ammonium salts and as soluble salts of organic bases, such as mono-, di- or triethanolamine. The anionic surfactants are preferably present in the form of their sodium or potassium salts, in particular in the form of the sodium salts.

The surfactants may be contained in the detergents or detergents according to the invention overall in an amount of preferably from 5% by weight to 50% by weight, in particular from 8% by weight to 30% by weight, based on the finished composition ,

Detergents or cleaners according to the invention may contain bleaches. Among the compounds which serve as bleaches and deliver H ₂ O ₂ in water, the sodium percarbonate, the sodium perborate tetrahydrate and the

Sodium perborate monohydrate particular importance. Other useful bleaching agents are, for example, peroxopyrophosphates, citrate perhydrates and H ₂ O ₂ -producing peracidic salts or peracids, such as persulfates or persulfuric acid. Also useful is the urea peroxohydrate percarbamide, which can be described by the formula H ₂ N-CO-NH ₂ H ₂ O ₂ . In particular, when using the means for cleaning hard surfaces, for example in automatic dishwashing, they may, if desired, also contain bleaching agents from the group of organic bleaches, although their use is also possible in principle for laundry detergents. Typical organic bleaches are the diacyl peroxides, such as dibenzoyl peroxide. Other typical organic bleaches are the peroxyacids, examples of which include the alkyl peroxyacids and the aryl peroxyacids. Preferred representatives are the peroxybenzoic acid and its ring-substituted derivatives, such as alkylperoxybenzoic acids, but also peroxy-α-naphthoic acid and magnesium monoperphthalate, the aliphatic or substituted aliphatic peroxyacids, such as peroxylauric acid, peroxystearic acid, ε-phthalimido- peroxycaproic acid (phthalimidoperoxyhexanoic acid, PAP), o-carboxybenzamidoperoxycaproic acid, N-nonenylamidoperadipic acid and N-nonylamidopersuccinates; and aliphatic and araliphatic peroxydicarboxylic acids such as 1,12-diperoxycarboxylic acid, 1,9-diperoxyazelaic acid, diperoxysebacic acid, diperoxybrassic acid, the diperoxyphthalic acids, 2 -Decyldiperoxybutane-1,4-diacid, N, N-terephthaloyl-di (6-aminopercaproic acid) can be used.

The content of bleach detergent or cleaning agent may be from 1 to 40% by weight and in particular from 10 to 20% by weight, with perborate monohydrate or percarbonate being advantageously used.

In order to achieve an improved bleaching effect during washing at temperatures of 60 ^° C. and below, and in particular during the laundry pretreatment, the agents may also contain bleach activators. As bleach activators, it is possible to use compounds which, under perhydrolysis conditions, give aliphatic peroxycarboxylic acids having preferably 1 to 10 C atoms, in particular 2 to 4 C atoms, and / or optionally substituted perbenzoic acid. Suitable substances are those which carry O- and / or N-acyl groups of the stated C atom number and / or optionally substituted benzoyl groups. Preference is given to polyacylated alkylenediamines, in particular tetraacetylethylenediamine (TAED), acylated triazine derivatives, in particular 1,5-diacetyl-2,4-dioxohexahydro-1,3,5-triazine (DADHT), acylated glycolurils, in particular 1, 3,4 , 6-tetraacetylglycoluril (TAGU), N-acylimides, in particular N-nonanoyl-succinimide (NOSI), acylated phenolsulfonates, especially n-nonanoyl or isononanoyloxybenzenesulfonate (n- or iso-NOBS), acylated hydroxycarboxylic acids, such as triethyl-O-acetyl citrate (TEOC), carboxylic anhydrides, in particular phthalic anhydride, isoic anhydride and / or succinic anhydride, carboxamides, such as N-methyldiacetamide, glycolide, acylated polyhydric alcohols, in particular triacetin, ethylene glycol diacetate, isopropenyl acetate, 2,5-diacetoxy-2,5-dihydrofuran and the from the German patent applications DE 19616693 and DE 19616767 known enol esters and acetylated sorbitol and mannitol or their in the European Patentanmeldu EP 0525239), acylated sugar derivatives, especially pentaacetylglucose (PAG), pentaacetylfructose, tetraacetylxylose and octaacetyllactose as well as acetylated, optionally N-alkylated glucamine or gluconolactone, triazole or triazole derivatives and / or particulate caprolactams and / or caprolactam derivatives, preferably N-acylated lactams, for example N-benzoylcaprolactam and N-acetylcaprolactam, which are described in International Patent Applications WO 94/27970, WO 94/28102, WO 94/28103 , WO 95/00626, WO 95/14759 and WO 95/17498. The hydrophilic substituted acyl acetals known from German patent application DE 19616769 and the acyllactams described in German patent application DE 19616770 and international patent application WO 95/14075 are likewise preferably used. The combinations of conventional bleach activators known from German patent application DE 4443177 can also be used. Likewise, nitrile derivatives such as cyanopyridines, nitrile quats, for example N-alkylammonium acetonitriles, and / or cyanamide derivatives can be used. Preferred bleach activators are sodium 4- (octanoyloxy) benzenesulfonate, π-nonanoyl or isononanoyloxybenzenesulfonate (n- or iso-NOBS), undecenoyloxybenzenesulfonate (UDOBS), sodium dodecanoyloxybenzenesulfonate (DOBS), decanoyloxybenzoic acid (DOBA, OBC 10) and / or Dodecanoyloxybenzenesulfonate (OBS 12), as well as N-methylmorpholinum acetonitrile (MMA). Such bleach activators can be used in the customary amount range of from 0.01 to 20% by weight, preferably in amounts of from 0.1 to 15% by weight, in particular from 1% to 10% by weight, based on the total composition, be included.

In addition to or in place of the conventional bleach activators, so-called bleach catalysts may also be included. These substances are bleach-enhancing transition metal salts or transition metal complexes such as, for example, Mn, Fe, Co, Ru or Mo salt complexes or carbonyl complexes. Mn, Fe, Co, Ru, Mo, Ti, V and Cu complexes with N-containing tripod ligands and Co, Fe, Cu and Ru ammine complexes are also suitable as bleach catalysts, wherein such compounds are preferably used, which are described in DE 19709284 A1.

Detergents or cleaners according to the invention generally comprise one or more builders, in particular zeolites, silicates, carbonates, organic cobuilders and, where there are no ecological reasons against their use, also the phosphates. The latter are particularly preferred builders to be used in automatic dishwashing detergents. To mention here are crystalline, layered sodium silicates of the general formula

NaMSi _x O _{2x + 1} yH ₂ O, where M is sodium or hydrogen, x is a number from 1.6 to 4, preferably 1.9 to 4.0 and y is a number from 0 to 20 and preferred values for x 2 , 3 or 4 are. Such crystalline layered silicates are described, for example, in European Patent Application EP 164514. Preferred crystalline layered silicates of the formula given are those in which M is sodium and x assumes the values 2 or 3. In particular, both β- and δ-sodium disilicates

Na ₂ Si ₂ O ₅ YH ₂ O is preferred. Such compounds are commercially available, for example, under the name ^SKS® (Clariant company). That is how it is

SKS-6 ^® mainly to a δ-sodium disilicate with the formula Na ₂ Si ₂ O ₅ -VH ₂ O, in SKS-7 ^® mainly to the ß-sodium disilicate. Reaction with acids (for example citric acid or carbonic acid) produces kanemite from the δ-sodium disilicate

NaHSi ₂ O ₅ -VH ₂ O, commercially available under the names SKS- ^9® or SKS- ^10® (Clariant). It may also be advantageous to use chemical modifications of these phyllosilicates. For example, the alkalinity of the layered silicates can be suitably influenced. Phyllosilicates doped with phosphate or with carbonate have altered crystal morphologies in comparison with the δ-sodium disilicate, dissolve more rapidly and show an increased calcium binding capacity in comparison to δ-sodium disilicate. Thus, phyllosilicates of the general empirical formula x Na ₂ O • y SiO ₂ • z P ₂ O ₅ , in which the ratio x to y is a number 0.35 to 0.6, the ratio x to z a number from 1, 75 to 1200 and the ratio y to z correspond to a number from 4 to 2800, described in the patent application DE 19601063. The solubility of the layered silicates can also be increased by using particularly finely divided layered silicates. Also compounds from the crystalline layer silicates with other ingredients can be used. In particular, compounds with cellulose derivatives which have advantages in the disintegrating effect and are used in particular in detergent tablets, and compounds with polycarboxylates, for example citric acid, or polymeric polycarboxylates, for example copolymers of acrylic acid, may be mentioned. It is also possible to use amorphous sodium silicates with a Na ₂ O: SiO ₂ modulus of from 1: 2 to 1: 3.3, preferably from 1: 2 to 1: 2.8 and in particular from 1: 2 to 1: 2.6, which Delayed and have secondary washing properties. The dissolution delay compared with conventional amorphous sodium silicates may have been caused in various ways, for example by surface treatment, compounding, compaction / densification or by overdrying. In the context of this invention, the term "amorphous" is also understood to mean "X-ray amorphous". This means that in X-ray diffraction experiments the silicates do not give sharp X-ray reflexes typical of crystalline substances, but at best one or more maxima of the scattered X-rays having a width of several degrees of diffraction angle. However, it may well even lead to particularly good builder properties if the silicate particles provide blurred or even sharp diffraction maxima in electron diffraction experiments. This is to be interpreted as meaning that the products have microcrystalline regions of size 10 to a few hundred nm, values of up to max. 50 nm and in particular up to max. 20 nm are preferred. Particularly preferred are compacted / compacted amorphous silicates, compounded amorphous silicates and overdried X-ray amorphous silicates.

An optionally usable, finely crystalline, synthetic and bound water-containing zeolite is preferably zeolite A and / or P. As zeolite P zeolite MAP ^® (commercial product from Crosfield) is particularly preferred. Also suitable, however, are zeolite X and mixtures of A, X and / or P. Commercially available and preferably usable in the context of the present invention is, for example, a cocrystal of zeolite X and zeolite A (about 80% by weight of zeolite X) ), which is sold by the company CONDEA Augusta SpA under the brand name VEGOBOND AX ^® and by the formula

n Na ₂ O ^• (1-H) ^■ K ₂ O Al ₂ O ₃ '(2 to 2.5) SiO ₂ ^■ (3.5 to 5.5) H ₂ O

can be described. Suitable zeolites have an average particle size of less than 10 μm (volume distribution, measuring method: Coulter Counter) and preferably contain 18 to 22% by weight, in particular 20 to 22% by weight, of bound water. Of course, a use of the well-known phosphates as builders is possible, unless such use should not be avoided for environmental reasons. Among the large number of commercially available phosphates, the alkali metal phosphates, with particular preference for pentasodium or pentapotassium triphosphate (sodium or potassium tripolyphosphate), are of greatest importance in the washing and cleaning agent industry.

Alkali metal phosphates is the summary term for the alkali metal (especially sodium and potassium) salts of various phosphoric acids, in which one can distinguish metaphosphoric acids (HPOs) _n and orthophosphoric H ₃ PO _{4 in} addition to high molecular weight representatives. The phosphates combine several advantages: they act as alkali carriers, prevent lime deposits on machine parts or lime incrustations in fabrics and also contribute to the cleaning performance.

Sodium dihydrogen phosphate, NaHaPO ₄ , exists as a dihydrate (density 1, 91 like ^"3 , melting point 60 °) and as a monohydrate (density 2.04 like ^{" 3} ). Both salts are white, very soluble in water powders that lose the water of crystallization on heating and at 200 ⁰ C in the weak acid diphosphate (disodium hydrogenated diphosphate, Na ₂ H ₂ P ₂ O ₇ ), at higher temperature in Natiumtrimetaphosphat (Na ₃ PsOg ) and Maddrell's salt (see below). NaH ₂ PO _{4 is} acidic; It arises when phosphoric acid is adjusted to a pH of 4.5 with sodium hydroxide solution and the mash is sprayed. Potassium dihydrogen phosphate (potassium phosphate primary or monobasic phosphate, potassium biphosphate, KDP), KH ₂ PO ₄ , is a white salt of density 2.33 ^"3 , has a melting point of 253 ⁰ C [decomposition to form potassium polyphosphate (KPO ₃ ) _X ] and is easily soluble in water.

Disodium hydrogen phosphate (secondary sodium phosphate), Na ₂ HPO ₄ , is a colorless, very slightly water-soluble crystalline salt. It exists anhydrous and with 2 mol. (Density 2.066 like ^'3 , loss of water at 95 °), 7 mol. (Density 1, 68 like ^"3 , melting point 48 ⁰ C with loss of 5 H ₂ O) and 12 mol (Density 1, 52 like ^"3 , melting point 35 ° C with loss of 5 H ₂ O), becomes anhydrous at 100 ⁰ C and goes stronger heating in the diphosphate Na ₄ P ₂ O ₇ over. Disodium hydrogen phosphate is prepared by neutralization of phosphoric acid with soda solution using phenolphthalein as an indicator. Dipotassium hydrogen phosphate (secondary or dibasic potassium phosphate), K ₂ HPO ₄ , is an amorphous, white salt that is readily soluble in water.

Trisodium phosphate, tertiary sodium phosphate, Na ₃ PO _4, colorless crystals, as the dodecahydrate a density of 1, 62 like ^"3 and a melting point of 73-76 ⁰ C (decomposition), as the decahydrate (corresponding to 19-20% P ₂ O ) has a melting point of 100 ⁰ C and in anhydrous form (corresponding to 39-40% P ₂ O ₅₎ like to have a density of 2.536 ^{'. 3} trisodium phosphate is readily soluble in water with an alkaline reaction and is prepared by evaporating a solution of exactly 1 mole of Tripotassium phosphate and 1 mole NaOH Tripotassium phosphate (tertiary or tribasic potassium phosphate), K ₃ PO ₄ , is a white, deliquescent, granular powder of density 2.56 ^"3 , has a melting point of 1340 ° and is in water with alkaline reaction easily soluble. It is produced, for example, by heating Thomasschlacke with coal and potassium sulfate. Despite the higher price, the more soluble, therefore highly effective, potassium phosphates are often preferred over the corresponding sodium compounds in the detergent industry.

Tetrasodium diphosphate (sodium pyrophosphate), Na ₄ P ₂ O ₇ , exists in anhydrous form (density 2.534 ^'3 , melting point 988 ° C, also indicated 88O ⁰ C) and as decahydrate (density 1, 815-1, 836 like ^"3 , Melting point 94 ° C. with loss of water.) Both substances are colorless crystals which are soluble in water with an alkaline reaction Na ₄ P ₂ O ₇ is formed on heating disodium phosphate to> 200 ° C. or by reacting phosphoric acid with soda in a stoichiometric ratio and the solution The decahydrate complexes heavy metal salts and hardness agents and therefore reduces the hardness of the water.Kali diphosphate (potassium pyrophosphate), K ₄ P ₂ O ₇ , exists in the form of the trihydrate and produces a colorless, hygroscopic powder with a density of 2.33 like ^"3 , which is soluble in water, wherein the pH of the 1% solution at 25 ° C is 10.4.

Condensation of NaH ₂ PO ₄ or KH ₂ PO ₄ gives rise to higher molecular weight sodium and potassium phosphates in which cyclic representatives, the sodium or potassium metaphosphates and chain types that can distinguish sodium and potassium polyphosphates, respectively. In particular, for the latter are a variety of names in use: hot or cold phosphates, Graham's salt, Kurrolsches and Maddrell's salt. All higher sodium and potassium phosphates are collectively referred to as condensed phosphates.

The industrially important pentasodium triphosphate (Na ₅ P ₃ O ₁₀ ; sodium tripolyphosphate) is an anhydrous or water-soluble salt of the general formula NaO- [P (O) (ONa) -O] _{n which} crystallizes with 6 H ₂ O and which is non-hygroscopic. Na with n = 3. In 100 g of water at room temperature dissolve about 17 g, at 6O ⁰ C about 20 g, at 100 ⁰ C, about 32 g of the salt water-free salt; after two hours of heating the solution to 100 ⁰ C caused by hydrolysis about 8% orthophosphate and 15% diphosphate. In the preparation of pentasodium triphosphate, phosphoric acid is reacted with soda solution or sodium hydroxide solution in a stoichiometric ratio and the solution is dehydrated by spraying. Similar to Graham's salt and sodium diphosphate, pentasodium triphosphate dissolves many insoluble metal compounds (including lime soaps, etc.). Pentakalium triphosphate, K5P ₃ O 10 (potassium tripolyphosphate), is commercially available, for example, in the form of a 50% by weight solution (> 23% P ₂ O ₅ , 25% K ₂ O). The potassium polyphosphates are widely used in the washing and cleaning industry. There are also sodium potassium tripolyphosphates which can also be used in the context of the present invention. These arise, for example, when hydrolyzed sodium trimetaphosphate with KOH:

(NaPOs) ₃ + 2 KOH -> Na ₃ K ₂ P ₃ Oi ₀ + H ₂ O

These are used according to the invention exactly as sodium tripolyphosphate, potassium tripolyphosphate or mixtures of these two; Mixtures of sodium tripolyphosphate and sodium potassium tripolyphosphate or mixtures of potassium tripolyphosphate and sodium potassium tripolyphosphate or mixtures of sodium tripolyphosphate and potassium tripolyphosphate and sodium potassium tripolyphosphate can also be used according to the invention. As organic cobuilders, it is possible in particular to use in the detergents and cleaners according to the invention polycarboxylates or polycarboxylic acids, polymeric polycarboxylates, polyaspartic acid, polyacetals, optionally oxidized dextrins, further organic cobuilders (see below) and phosphonates. These classes of substances are described below.

Useful organic builder substances are, for example, the polycarboxylic acids which can be used in the form of their sodium salts, polycarboxylic acids meaning those carboxylic acids which carry more than one acid function. These are, for example, citric acid, adipic acid, succinic acid, glutaric acid, malic acid, tartaric acid, maleic acid, fumaric acid, sugar acids, aminocarboxylic acids, nitrilotriacetic acid (NTA), if such use can not be avoided for ecological reasons, and mixtures of these. Preferred salts are the salts of polycarboxylic acids such as citric acid, adipic acid, succinic acid, glutaric acid, tartaric acid, sugar acids and mixtures thereof.

The acids themselves can also be used. In addition to their builder effect, they also typically have the property of an acidifying component and thus also serve to set a lower and milder pH of detergents or cleaners, unless the pH resulting from the mixture of the other components is desired. In particular, environmentally friendly acids such as citric acid, acetic acid, tartaric acid, malic acid, lactic acid, glycolic acid, succinic acid, glutaric acid, adipic acid, gluconic acid and any mixtures of these are to be mentioned here. But also mineral acids, in particular sulfuric acid or bases, in particular ammonium or alkali hydroxides can serve as pH regulators. Such regulators are contained in the agents according to the invention in amounts of preferably not more than 20% by weight, in particular from 1.2% by weight to 17% by weight.

Further suitable builders are polymeric polycarboxylates, for example the alkali metal salts of polyacrylic acid or polymethacrylic acid, for example those having a relative molecular mass of from 500 to 70,000 g / mol. For the purposes of this document, the molecular weights stated for polymeric polycarboxylates are weight-average molar masses M _{w of} the particular acid form, which were determined in principle by means of gel permeation chromatography (GPC), a UV detector being used. The measurement was carried out against an external polyacrylic acid standard, which provides realistic molecular weight values due to its structural relationship with the polymers investigated. These data differ significantly from the molecular weight data, in which polystyrene sulfonic acids are used as standard. The molar masses measured against polystyrenesulfonic acids are generally significantly higher than the molecular weights specified in this document.

Suitable polymers are, in particular, polyacrylates which preferably have a molecular weight of 2,000 to 20,000 g / mol. Because of their superior solubility, the short-chain polyacrylates, which have molar masses of from 2000 to 10000 g / mol, and particularly preferably from 3000 to 5000 g / mol, may again be preferred from this group.

Also suitable are copolymeric polycarboxylates, in particular those of acrylic acid with methacrylic acid and of acrylic acid or methacrylic acid with maleic acid. Copolymers of acrylic acid with maleic acid which contain 50 to 90% by weight of acrylic acid and 50 to 10% by weight of maleic acid have proven to be particularly suitable. Their relative molecular weight, based on free acids, is generally from 2000 to 70000 g / mol, preferably from 20,000 to 50,000 g / mol and in particular from 30,000 to 40,000 g / mol. The (co) polymeric polycarboxylates can be used either as a powder or as an aqueous solution. The content of the (co) polymeric polycarboxylates may be from 0.5 to 20% by weight, in particular from 1 to 10% by weight.

To improve the water solubility, the polymers may also contain allylsulfonic acids such as allyloxybenzenesulfonic acid and methallylsulfonic acid as a monomer.

Also particularly preferred are biodegradable polymers of more than two different monomer units, for example those which are salts of the monomers Acrylic acid and maleic acid and vinyl alcohol or vinyl alcohol derivatives or containing as monomers salts of acrylic acid and 2-alkylallylsulfonic acid and sugar derivatives.

Further preferred copolymers are those which have as monomers preferably acrolein and acrylic acid / acrylic acid salts or acrolein and vinyl acetate.

Also to be mentioned as further preferred builders polymeric aminodicarboxylic acids, their salts or their precursors. Particular preference is given to polyaspartic acids or their salts and derivatives.

Further suitable builder substances are polyacetals which can be obtained by reacting dialdehydes with polyolcarboxylic acids which have 5 to 7 C atoms and at least 3 hydroxyl groups. Preferred polyacetals are obtained from dialdehydes such as glyoxal, glutaraldehyde, terephthalaldehyde and mixtures thereof and from polyol carboxylic acids such as gluconic acid and / or glucoheptonic acid.

Further suitable organic builder substances are dextrins, for example oligomers or polymers of carbohydrates, which can be obtained by partial hydrolysis of starches. The hydrolysis can be carried out by customary, for example acid or enzyme catalyzed processes. Preferably, it is hydrolysis products having average molecular weights in the range of 400 to 500,000 g / mol. In this case, a polysaccharide with a dextrose equivalent (DE) in the range from 0.5 to 40, in particular from 2 to 30 is preferred, DE being a customary measure of the reducing action of a polysaccharide compared to dextrose, which is a DE of 100 has. Usable are both maltodextrins with a DE between 3 and 20 and dry glucose syrups with a DE between 20 and 37 and so-called yellow dextrins and white dextrins with higher molecular weights in the range from 2000 to 30,000 g / mol.

The oxidized derivatives of such dextrins are their reaction products with oxidizing agents which are capable of oxidizing at least one alcohol function of the saccharide ring to the carboxylic acid function. Especially Preferred organic builders for agents according to the invention are oxidized starches or their derivatives from the applications EP 472042, WO 97/25399, and EP 755944.

Oxydisuccinates and other derivatives of disuccinates, preferably ethylenediamine disuccinate, are other suitable co-builders. In this case, ethylenediamine-N, ^{N'-disuccinate} (EDDS) is preferably in the form of its sodium or magnesium salts. Also preferred in this context are glycerol disuccinates and glycerol trisuccinates. Suitable amounts are in zeolith-, carbonate and / or silicate-containing formulations between 3 and 15 wt .-%.

Other useful organic cobuilders are, for example, acetylated hydroxycarboxylic acids or their salts, which may optionally also be present in lactone form and which contain at least 4 carbon atoms and at least one hydroxyl group and a maximum of two acid groups.

Another substance class with cobuilder properties are the phosphonates. These are, in particular, hydroxyalkane or aminoalkanephosphonates. Among the hydroxyalkane phosphonates, the 1-hydroxyethane-1, 1-diphosphonate (HEDP) is of particular importance as a cobuilder. It is preferably used as the sodium salt, the disodium salt neutral and the tetrasodium salt alkaline (pH 9). Preferred aminoalkanephosphonates are ethylenediamine tetramethylenephosphonate (EDTMP), diethylenetriaminepentamethylenephosphonate (DTPMP) and their higher homologs. They are preferably used in the form of the neutral-reacting sodium salts, for example as the hexasodium salt of EDTMP or as the hepta- and octa-sodium salt of DTPMP. The builder used here is preferably HEDP from the class of phosphonates. The aminoalkanephosphonates also have a pronounced heavy metal binding capacity. Accordingly, in particular if the agents also contain bleach, it may be preferable to use aminoalkanephosphonates, in particular DTPMP, or to use mixtures of the phosphonates mentioned. Moreover, all compounds capable of forming complexes with alkaline earth ions can be used as co-builders.

Builder substances may optionally be present in the detergents or cleaners according to the invention in amounts of up to 90% by weight. They are preferably contained in amounts of up to 75% by weight. Detergents according to the invention have builder contents of, in particular, from 5% by weight to 50% by weight. In agents according to the invention for the cleaning of hard surfaces, in particular for the automated cleaning of dishes, the content of builder substances is in particular from 5% by weight to 88% by weight, wherein preferably no water-insoluble builder materials are used in such agents. In a preferred embodiment of the invention means for the particular automatic cleaning of dishes are 20 wt .-% to 40 wt .-% of water-soluble organic builder, in particular alkali, 5 wt .-% to 15 wt .-% alkali carbonate and 20 wt .-% bis 40 wt .-% Alkalidisilikat included.

Solvents that can be used in the liquid to gelatinous compositions of detergents and cleaners, for example, from the group of monohydric or polyhydric alcohols, alkanolamines or glycol ethers, provided that they are miscible in the specified concentration range with water. The solvents are preferably selected from ethanol, n- or i-propanol, butanols, ethylene glycol methyl ether, ethylene glycol ethyl ether, ethylene glycol propyl ether,

Ethylene glycol mono-n-butyl ether, diethylene glycol methyl ether, diethylene glycol ethyl ether, propylene glycol methyl, ethyl or propyl ether, dipropylene glycol monomethyl, or ethyl ether, di-isopropylene glycol monomethyl, or ethyl ether, methoxy, ethoxy or butoxy triglycol, 1 Butoxyethoxy-2-propanol, 3-methyl-3-methoxybutanol, propylene glycol t-butyl ether and mixtures of these solvents.

Solvents may be used in the liquid to gelled detergents and cleaners according to the invention in amounts of between 0.1 and 20% by weight, but preferably below 15% by weight and in particular below 10% by weight.

To adjust the viscosity, one or more thickeners or thickening systems can be added to the composition according to the invention. These high-molecular substances, which are also called swelling agents, usually absorb the liquids and swell up to finally pass into viscous true or colloidal solutions.

Suitable thickeners are inorganic or polymeric organic compounds. The inorganic thickeners include, for example, polysilicic acids, clay minerals such as montmorillonites, zeolites, silicas and bentonites. The organic thickeners are derived from the groups of natural polymers, modified natural polymers and fully synthetic polymers. Such naturally derived polymers include, for example, agar-agar, carrageenan, tragacanth, gum arabic, alginates, pectins, polyoses, guar gum, locust bean gum, starch, dextrins, gelatin and casein. Modified natural products, which are used as thickeners, come mainly from the group of modified starches and celluloses. By way of example, carboxymethylcellulose and other cellulose ethers, hydroxyethyl and propylcellulose and core flour ethers may be mentioned here. Fully synthetic thickeners are polymers such as polyacrylic and polymethacrylic compounds, vinyl polymers, polycarboxylic acids, polyethers, polyimines, polyamides and polyurethanes.

The thickeners may be present in an amount of up to 5% by weight, preferably from 0.05 to 2% by weight, and more preferably from 0.1 to 1.5% by weight, based on the finished composition ,

Detergents according to the invention may contain, as optical brighteners, derivatives of diaminostilbenedisulfonic acid or their alkali metal salts. For example, salts of 4,4'-bis (2-anilino-4-morpholino-1, 3,5-triazinyl-6-amino) stilbene-2,2'-disulphonic acid or similarly constructed compounds which replace the morpholino Group carry a diethanolamino group, a methylamino group, an anilino group or a 2-methoxyethylamino group. Further, brighteners of the substituted diphenylstyrene type may be present, for example, the alkali salts of 4,4'-bis (2-sulfostyryl) -diphenyl, 4,4'-bis (4-chloro-3-sulfostyryl) -diphenyl, or 4 - (4-chlorostyryl) -4 '- (2-sulfostyryl). Mixtures of the aforementioned optical brightener can be used.

Graying inhibitors have the task of being detached from the textile fiber Keep dirt suspended in the fleet. Water-soluble colloids of mostly organic nature are suitable for this purpose, for example starch, glue, gelatin, salts of ether carboxylic acids or ether sulfonic acids of starch or of cellulose or salts of acidic sulfuric acid esters of cellulose or starch. Also, water-soluble polyamides containing acidic groups are suitable for this purpose. Furthermore, other than the above-mentioned starch derivatives can be used, for example aldehyde starches. Preference is given to using cellulose ethers, such as carboxymethylcellulose (Na salt), methylcellulose, hydroxyalkylcellulose and mixed ethers, such as methylhydroxyethylcellulose, methylhydroxypropylcellulose, methylcarboxymethylcellulose and mixtures thereof, for example in amounts of from 0.1 to 5% by weight, based on the compositions ,

To effect silver corrosion protection, silver corrosion inhibitors can be used in dishwashing detergents according to the invention. Such are known from the prior art, for example benzotriazoles, iron (III) chloride or CoSO ₄ . For example, as known from European Patent EP 0 736 084 B1, particularly suitable silver corrosion inhibitors for use in conjunction with enzymes are manganese, titanium, zirconium, hafnium, vanadium, cobalt or cerium salts and / or complexes where the said metals are present in one of the oxidation states II, III, IV, V or VI. Examples of such compounds are MnSO ₄ , V ₂ O ₅ , V ₂ O ₄ , VO ₂ , TiOSO ₄ , K ₂ TiF ₆ , K ₂ ZrF ₆ , Co (NO ₃ ) ₂ , Co (NO ₃ ) ₃ , and the like mixtures.

"Soil-release" or "soil repellents" are mostly polymers which impart soil repellency when used in a laundry detergent detergent and / or aid in the soil release performance of the other detergent ingredients. A similar effect can also be observed in their use in hard surface cleaners.

Particularly effective and long-known soil release agents are copolyesters with dicarboxylic acid, alkylene glycol and polyalkylene glycol units. Examples thereof are copolymers or copolymers of polyethylene terephthalate and polyoxyethylene glycol (DT 16 17 141, or DT 22 OO 911). In the German patent application DT 22 53 063 acidic agents are mentioned, including a Copolymer of a dibasic carboxylic acid and an alkylene or cycloalkylene polyglycol. Polymers of ethylene terephthalate and polyethylene oxide terephthalate and their use in detergents are described in German patents DE 28 57 292 and DE 33 24 258 and European patent EP 0 253 567. European patent EP 066 944 relates to compositions containing a copolyester of ethylene glycol, polyethylene glycol, aromatic dicarboxylic acid and sulfonated aromatic dicarboxylic acid in certain molar ratios. European Patent EP 0 185 427 discloses methyl or ethyl group-end capped polyesters having ethylene and / or propylene terephthalate and polyethylene oxide terephthalate units and laundry detergents containing such soil release polymer. European patent EP 0 241 984 relates to a polyester which, besides oxyethylene groups and terephthalic acid units, also contains substituted ethylene units and also glycerine units. European Patent EP 0 241 985 discloses polyesters which, in addition to oxyethylene groups and terephthalic acid units, contain 1, 2-propylene, 1, 2-butylene and / or 3-methoxy-1, 2-propylene groups and also glycerol units and with C 1 - Are capped to C ₄ alkyl groups. _-4 alkyl or acyl groups end-capped polyesters with polypropylene terephthalate and polyoxyethylene terephthalate units are at least in part by C from the European patent application EP 0 272 033.. European Patent EP 0 274 907 describes sulfoethyl end-capped terephthalate-containing soil release polyesters. According to European Patent Application EP 0 357 280, sulfonation of unsaturated end groups produces soil release polyesters with terephthalate, alkylene glycol and poly-C _2-4 glycol units. International Patent Application WO 95/32232 relates to acidic, aromatic soil release polymers. International Patent Application WO 97/31085 discloses non-polymeric soil repellent active ingredients for multi-functional cotton materials: a first entity, which may be cationic, for example, is capable of adsorption to the cotton surface by electrostatic interaction, and a second Unit that is hydrophobic is responsible for the retention of the drug at the water / cotton interface.

The color transfer inhibitors which are suitable for use in textile detergents according to the invention include, in particular, polyvinylpyrrolidones, polyvinyl imidazoles, polymeric N-oxides such as poly (vinylpyridine N-oxide) and copolymers of vinylpyrrolidone with vinylimidazole.

When used in automated cleaning processes, it may be advantageous to add foam inhibitors to the agents concerned. As foam inhibitors are, for example, soaps of natural or synthetic origin, which have a high proportion of Ci ₈ -C24 fatty acids. Suitable non-surfactant foam inhibitors are, for example, organopolysiloxanes and mixtures thereof with microfine, optionally signed silica and paraffins, waxes, microcrystalline waxes and mixtures thereof with signed silica or bistearylethylenediamide. It is also advantageous to use mixtures of various foam inhibitors, for example those of silicones, paraffins or waxes. Preferably, the foam inhibitors, in particular silicone and / or paraffin-containing foam inhibitors, are bound to a granular, water-soluble, or dispersible carrier substance. In particular, mixtures of paraffins and bistearylethylene diamides are preferred.

In addition, a hard surface cleaning agent according to the invention may contain abrasive constituents, in particular from the group comprising quartz flours, wood flours, plastic flours, chalks and glass microspheres and mixtures thereof. Abrasive substances are preferably not more than 20% by weight, in particular from 5% by weight to 15% by weight, in the cleaning agents according to the invention.

Dyes and fragrances are added to detergents and cleaners in order to improve the aesthetic appearance of the products and to provide the consumer with a visually and sensory "typical and unmistakable" product in addition to the washing and cleaning performance. As perfume oils or fragrances, individual perfume compounds, for example the synthetic products of the ester type, ethers, aldehydes, ketones, alcohols and hydrocarbons can be used. Fragrance compounds of the ester type are, for example, benzyl acetate, phenoxyethyl isobutyrate, p-tert-butylcyclohexyl acetate, linalyl acetate, dimethylbenzylcarbinyl acetate, phenylethyl acetate, linalyl benzoate, benzyl formate, ethylmethylphenyl glycinate, allylcyclohexyl propionate, styrallyl propionate and benzyl salicylate. The ethers include, for example, benzyl ethyl ethers, to the Aldehydes, for example, the linear alkanals having 8-18 C atoms, citral, citronellal, citronellyloxyacetaldehyde, cyclamen aldehyde, hydroxycitronellal, lilial and bourgeonal, to the ketones for example the ionone, α-isomethylionone and methyl cedrylketone, to the alcohols anethole, citronellol , Eugenol, geraniol, linalool, phenylethyl alcohol and terpineol, the hydrocarbons mainly include the terpenes such as limonene and pinene. Preferably, however, mixtures of different fragrances are used, which together produce an attractive fragrance. Such perfume oils may also contain natural fragrance mixtures such as are available from vegetable sources, for example, pine, citrus, jasmine, patchouly, rose or ylang-ylang oil. Also suitable are muscatel, sage, chamomile, clove, lemon balm, mint, cinnamon, lime, juniper, vetiver, olibanum, galbanum and labdanum, and orange blossom, neroliol, orange peel and sandalwood. Usually, the content of detergents and cleaners to dyes is less than 0.01 wt .-%, while perfumes can account for up to 2% by weight of the total formulation.

The fragrances can be incorporated directly into the detergents or cleaners, but it can also be advantageous to apply the fragrances to carriers, which enhance the adhesion of the perfume to the items to be cleaned and provide a slower release of fragrance for long-lasting fragrance, especially of treated textiles. As such carrier materials, for example, cyclodextrins have been proven, the cyclodextrin-perfume complexes can be additionally coated with other excipients. A further preferred carrier for fragrances is the described zeolite X, which can also absorb fragrances instead of or in mixture with surfactants. Preference is therefore given to washing and cleaning agents containing the described zeolite X and fragrances, which are preferably at least partially absorbed on the zeolite.

Preferred dyes, the selection of which presents no difficulty to the skilled person, have a high storage stability and insensitivity to the other ingredients of the agents and to light and no pronounced substantivity to textile fibers so as not to stain them. Detergents or cleaners may contain antimicrobial agents to combat microorganisms. Depending on the antimicrobial spectrum and mechanism of action, a distinction is made between bacteriostatic agents and bactericides, fungistatics and fungicides, etc. Important substances from these groups are, for example, benzalkonium chlorides, alkylarylsulfonates, halophenols and phenolmercuric acetate. The terms antimicrobial action and antimicrobial active substance have the usual meaning within the scope of the teaching according to the invention, which is described, for example, by KH Wallhäußer in "Praxis der Sterilisation, Disinfection - Conservation: Germ Identification - Company Hygiene" (5th edition - Stuttgart, New York: Thieme, 1995 ), wherein all the substances described there can be used with antimicrobial activity. Suitable antimicrobial agents are preferably selected from the groups of alcohols, amines, aldehydes, antimicrobial acids or their salts, carboxylic esters, acid amides, phenols, phenol derivatives, diphenyls, diphenylalkanes, urea derivatives, oxygen, nitrogen acetals and formals, benzamidines, isothiazolines, Phthalimide derivatives, pyridine derivatives, antimicrobial surface active compounds, guanidines, antimicrobial amphoteric compounds, quinolines, 1, 2-dibromo-2,4-dicyanobutane, iodo-2-propyl-butyl-carbamate, iodine, iodophores, peroxo compounds, halogen compounds and any mixtures of the foregoing ,

The antimicrobial agent may be selected from ethanol, n-propanol, i-propanol, 1,3-butanediol, phenoxyethanol, 1,2-propylene glycol, glycerol, undecylenic acid, benzoic acid, salicylic acid, dihydracetic acid, o-phenylphenol, N-methylmorpholine. acetonitrile (MMA), 2-benzyl-4-chlorophenol, 2,2'-methylenebis (6-bromo-4-chlorophenol), 4,4'-dichloro-2'-hydroxydiphenyl ether (dichlosan), 2,4 , 4'-trichloro-2'-hydroxydiphenyl ether (trichlosan), chlorhexidine, N- (4-chlorophenyl) -N- (3,4-dichlorophenyl) -urea, N, N '- (1, 10-decanediyldi- 1-pyridinyl-4-ylidene) bis- (1-octanamine) dihydrochloride, N, N'-bis (4-chlorophenyl) -3,12-diimino-2,4,11,13-tetraaza-tetradecane diimidamide, glucoprotamines, antimicrobial quaternary surface active compounds, guanidines including the bi- and polyguanidines such as 1,6-bis (2-ethylhexyl-biguanido-hexane) dihydrochloride, 1,6-di- (Ni, Ni '- phenyldiguanidone-N ₅ , N ₅ ') -hexane-tetrahydrochloride, 1,6-di- (Ni, Ni'-phenyl-Ni.Ni-methyldiguanido-N _δ .Ns') - hexane dihydrochloride, 1,6-di- (Ni, Ni'-o-chlorophenyldiguanido- N ₅ , N ₅ ') -hexane dihydrochloride, 1,6-di- (Ni, Ni') 2,6-dichlorophenyldiguanido-N ₅ , N ₅ ') hexane dihydrochloride, 1, 6-di- [N ₁ , Ni'-beta- (p-methoxyphenyl) diguanido-N ₅ , N ₅ '] -hexanes-chlorohydrochloride, 1,6-di- (N 1 -Ni'-alpha-methyl) .beta.-phenyldiguanido-Ns.Ns'J-hexane dihydrochloride, 1, 6-di-

(Ni, Ni'-p-nitrophenyldiguanido-N ₅ , N ₅ ') hexane dihydrochloride, omega: omega-di (Ni, Ni'-phenyldiguanido-N ₅ , N ₅ ') di-n-propyl ether dihydrochloride , omega: omega'-di (Ni, Ni'-p-chlorophenyldiguanido-N ₅ , N ₅ ; -di-n-propyl ether tetrahydrochloride, 1,6-di- (Ni, Ni'-2,4-dichlorophenyldiguanido -N ₅ , N ₅ ') hexane-tetrahydrochloride, 1,6-di- (Ni, N-i'-p-methylphenyldiguanido- N ₅ , N ₅ ') hexane dihydrochloride, 1,6-di- (Ni, Ni'-2,4, 5-trichlorophenyldiguanido-N ₅ , N ₅ ') hexane-tetrahydrochloride, 1,6-di- [Ni, Ni'-alpha (p-chlorophenyl) ethyldiguanido-N ₅ , N ₅ '] hexane dihydrochloride, omega: omega-di- (Ni, Ni'-p-chlorophenyldiguanido-N ₅ , N ₅ ') m-xylene dihydrochloride, 1, 12-di- (Ni, Ni'-p-chlorophenyldiguanido-N ₅ , N ₅ ') dodecane dihydrochloride, 1, 10-di- (Ni, N-P-phenyldiguanido- N ₅ , N ₅ ') -decane tetrahydrochloride, 1, 12-di- (Ni, Ni'-phenyldiguanido- N ₅₁ N ₅ ') dodecane-tetrahydrochloride, 1,6-di- (Ni, Ni'-o-chlorophenyldiguanido- N ₅₁ N ₅ ') hexane dihydrochloride, 1,6-di- (Ni, Ni'-o-chlorophene yldiguanido- N ₅ , N ₅ ') hexanetrahydrochloride, ethylene bis (1-tolyl biguanide), ethylene bis (p-tolyl biguanide), ethylene bis (3,5-dimethylphenyl biguanide), ethylene bis - (p-tert-amylphenylbiguanide), ethylene bis (nonylphenylbiguanide), ethylene bis (phenylbiguanide), ethylene bis (N-butylphenylbiguanide), ethylene bis (2,5-diethoxyphenylbiguanide), ethylene bis ( 2,4-dimethylphenyl biguanide), ethylene bis (o-diphenylbiguanide), ethylene bis (mixed amyl naphthyl biguanide), N-butyl ethylene bis (phenylbiguanide), trimethylene bis (o-tolyl biguanide), N-butyl trimethyl-bis (phenyl biguanides) and the corresponding salts such as acetates, gluconates, hydrochlorides, hydrobromides, citrates, bisulfites, fluorides, polymaleates, N-cocoalkyl sarcosinates, phosphites, hypophosphites, perfluorooctanoates, silicates, sorbates, salicylates, maleates, tartrates, fumarates , Ethylenediaminetetraacetate, Iminodiacetate, Cinnamate, Thiocyanate, Arginate, Pyromellitate, Tetracarboxybutyrate, Benzoate, Glutarate, Monofluorophosphate, Per fluoropropionates and any mixtures thereof. Also suitable are halogenated xylene and cresol derivatives, such as p-chloromethacresol or p-chloro-meta-xylene, and natural antimicrobial agents of plant origin (for example, from spices or herbs), of animal and microbial origin. Preferably, antimicrobial surface active quaternary compounds, a natural antimicrobial agent of plant origin and / or a natural antimicrobial agent of animal origin, most preferably at least one natural antimicrobial agent of plant origin from the group comprising caffeine, theobromine and theophylline and essential oils such Eugenol, thymol and geraniol, and / or at least one natural antimicrobial agent of animal origin from the group, comprising enzymes such as protein from milk, lysozyme and lactoperoxidase, and / or at least one antimicrobial surface active quaternary compound having an ammonium, sulfonium, phosphonium -, iodonium or Arsoniumgruppe, peroxo compounds and Chlorver¬ compounds are used. Also substances of microbial origin, so-called bacteriocins, can be used.

The suitable as antimicrobial agents quaternary ammonium compounds (QAV) have the general formula (R ¹ ) (R ² ) (R ³ ) (R ⁴ ) N ⁺ X ^" , in the R ¹ to R ⁴ identical or different CtC ₂₂ alkyl radicals , C ₇ -C ₂₈ -Aralkylreste or heterocyclic radicals, wherein two or in the case of an aromatic inclusion as in pyridine even three radicals together with the nitrogen atom, the heterocycle, for example a pyridinium or imidazolinium compound, form, and X ^" halide ions, sulfate ions , Hydroxide ions or similar anions. For optimum antimicrobial activity, preferably at least one of the radicals has a chain length of 8 to 18, in particular 12 to 16, carbon atoms.

QACs can be prepared by reacting tertiary amines with alkylating agents, such as, for example, methyl chloride, benzyl chloride, dimethyl sulfate, dodecyl bromide, but also ethylene oxide. The alkylation of tertiary amines with a long alkyl radical and two methyl groups succeeds particularly easily, and the quaternization of tertiary amines with two long radicals and one methyl group can be carried out with the aid of methyl chloride under mild conditions. Amines which have three long alkyl radicals or hydroxy-substituted alkyl radicals are less reactive and are preferably quaternized with dimethyl sulfate.

Suitable QACs are, for example, benzalkonium chloride (N-alkyl-N, N-dimethylbenzylammonium chloride, CAS No. 8001-54-5), benzalkone B (m, p-dichlorobenzyl-dimethyl-C 12 -alkylammonium chloride, CAS No. 58390- 78-6), benzoxonium chloride (benzyldodecyl bis (2-hydroxyethyl) ammonium chloride), cetrimonium bromide (N-hexadecyl-N, N-trimethylammonium bromide, CAS No. 57-09-0), benzetonium chloride ( N, N-dimethyl-N- [2- [2- [p- (1, 1, 3,3-tetramethylbutyl) phenoxy] ethoxy] ethyl] benzyl ammonium chloride, CAS No. 121-54-0), Dialkyldimethylammonium chlorides such as di-A7-decyl-dimethyl ammonium chloride (CAS No. 7173-51-5-5), didecyldi-methylammonium bromide (CAS No. 2390-68-3), dioctyl-dimethyl-ammonium chloride, 1-cetylpyridinium chloride (CAS No. 123-03-5) and thiazolin-iodide ( CAS No. 15764-48-1) and mixtures thereof. Particularly preferred QUATS are the benzalkonium chlorides ₈ alkyl radicals with C ₈ -C, especially Cia-C ^ -Aklyl-benzyl-dimethyl-ammonium chloride.

Benzalkonium halides and / or substituted benzalkonium halides are for example commercially available as Barquat ^® ex Lonza, Marquat® ^® ex Mason, Variquat ^® ex Witco / Sherex and Hyamine ^® ex Lonza and as Bardac ^® ex Lonza. Other commercially obtainable antimicrobial agents are N- (3-chloroallyl) - hexaminiumchlorid as Dowicide and Dowicil ^{® ®} ex Dow, benzethonium chloride such as Hyamine ^® 1622 ex Rohm & Haas, methylbenzethonium as Hyamine ^® 1ox ex Rohm & Haas, cetylpyridinium chloride such as Cepacol ex Merrell Labs ,

The antimicrobial agents are used in amounts of 0.0001 wt .-% to 1 wt .-%, preferably from 0.001 wt .-% to 0.8 wt .-%, particularly preferably from 0.005 wt .-% to 0.3 wt .-% and in particular from 0.01 to 0.2 wt .-% used.

The detergents or cleaners according to the invention may contain UV absorbents (UV absorbers) which are applied to the treated textiles and improve the lightfastness of the fibers and / or the lightfastness of other formulation components. Under UV absorber are organic substances (sunscreen) to understand, which are able to absorb ultraviolet rays and the absorbed energy in the form of longer-wave radiation, for example, to give off heat.

Compounds having these desired properties include, for example, the non-radiative deactivating compounds and derivatives of benzophenone having substituents in the 2- and / or 4-position. Furthermore, substituted benzotriazoles, in the 3-position phenyl-substituted acrylates (cinnamic acid derivatives, optionally with cyano groups in the 2-position), salicylates, organic Ni complexes and natural products such as Umbeiliferon and the body's own Urocansäure suitable. Of particular importance are biphenyl and especially Stilbenderivate as described for example in EP 0728749 A and are commercially available as Tinosorb ^® FD or Tinosorb ^® FR ex Ciba. As UV-B absorbers may be mentioned: 3-Benzylidencampher or 3-Benzylidennorcampher and its derivatives, for example 3- (4-Methylbenzy- liden) camphor, as described in EP 0693471 B1; 4-aminobenzoic acid derivatives, preferably 2-ethylhexyl 4- (dimethylamino) benzoate, A-

(Dimethylamino) benzoic acid 2-octyl ester and A-

(Dimethylamino) benzoesäureamylester; Esters of cinnamic acid, preferably 2-ethylhexyl A-methoxycinnamate, propyl 4-methoxycinnamate, isoamyl 4-methoxycinnamate, 2-cyano-3,3-phenylcinnamic acid 2-ethylhexyl ester (octocrylene); Esters of salicylic acid, preferably 2-ethylhexyl salicylate, 4-isopropylbenzyl salicylate, homomenthyl salicylate; Derivatives of benzophenone, preferably 2-hydroxy-4-methoxybenzophenone, 2-hydroxy-4-methoxy-4'-methylbenzophenone, 2,2'-dihydroxy-4-methoxybenzophenone; Esters of benzalmalonic acid, preferably di-2-ethylhexyl 4-methoxybenzmalonate; Triazine derivatives such as 2,4,6-trianilino- (p-carbo-2'-ethyl-1 '-hexyloxy) -1, 3,5-triazine and octyl triazone as described in EP 0818450 A1 or dioctyl butamido Triazone (Uvasorb® HEB); Propane-1,3-diones such as 1- (4-tert-butylphenyl) -3- (4'-methoxyphenyl) propane-1,3-dione; Ketotricyclo (5.2.1.0) decane derivatives, as described in EP 0694521 B1. Also suitable are 2-phenylbenzimidazole-5-sulfonic acid and its alkali metal, alkaline earth metal, ammonium, alkylammonium, alkanolammonium and glucammonium salts; Sulfonic acid derivatives of benzophenones, preferably 2-hydroxy-4-methoxybenzophenone-5-sulfonic acid and its salts; Sulfonic acid derivatives of 3-Benzylidencamphers, such as 4- (2-oxo-3-bomylidenemethyl) benzenesulfonic acid and 2-methyl-5- (2-oxo-3-bomylidene) sulfonic acid and salts thereof.

As a typical UV-A filter in particular derivatives of benzoylmethane come into question, such as 1- (4'-tert-butylphenyl) -3- (4'-methoxyphenyl) propane-1, 3-dione, 4-tert-butyl 4'-methoxydibenzoylmethane (Parsol 1789), 1-phenyl-3- (4'-isopropylphenyl) propane-1, 3-dione and also enamine compounds, as described in DE 19712033 A1 (BASF). Of course, the UV-A and UV-B filters can also be used in mixtures. In addition to the soluble substances mentioned, insoluble photoprotective pigments, namely finely dispersed, preferably nano-metal oxides or salts, are also suitable for this purpose. Examples of suitable metal oxides are in particular zinc oxide and titanium dioxide and, in addition, oxides of iron, zirconium, silicon, manganese, aluminum and cerium, and mixtures thereof. As salts silicates (talc), barium sulfate or zinc stearate can be used. The oxides and salts are already used in the form of the pigments for skin-care and skin-protecting emulsions and decorative cosmetics. The particles should have an average diameter of less than 100 nm, preferably between 5 and 50 nm and in particular between 15 and 30 nm. They may have a spherical shape, but it is also possible to use those particles which have an ellipsoidal or otherwise deviating shape from the spherical shape. The pigments may also be surface-treated, that is to say hydrophilized or hydrophobicized. Typical examples are coated titanium dioxides, for example titanium dioxide T 805 (Degussa) or Eusolex® T2000 (Merck; preferred hydrophilic coating agents are silicones and particularly preferably trialkoxyoctylsilanes or simethicones.) Micronized zinc oxide is preferably used see the review by P. Finkel in SÖFW Journal 122 (1996), p. 543.

The UV absorbents are usually used in amounts of from 0.01% by weight to 5% by weight, preferably from 0.03% by weight to 1% by weight.

A polypeptide according to the invention and / or other proteins contained can be protected especially during storage by stabilizers, for example from denaturation, disintegration or inactivation, for example by physical influences, oxidation or proteolytic cleavage. This applies to all agents according to the invention, in particular washing and cleaning agents.

One group of stabilizers are reversible protease inhibitors. Benzamidine hydrochloride, borax, boric acids, boronic acids or their salts or esters are frequently used for this purpose, including, in particular, derivatives with aromatic groups, for example ortho, meta or para-substituted phenylboronic acids, in particular A-formyl-biphenyl-boronic acid, or the salts or Esters of the compounds mentioned. Also peptide aldehydes, that is, oligopeptides with reduced C-terminus, in particular those of 2 to 50 monomers are used for this purpose. The peptidic reversible protease inhibitors include, among others Ovomucoid and Leupeptin. Also, specific, reversible peptide inhibitors for the protease subtilisin and fusion proteins from proteases and specific peptide inhibitors are suitable.

Other enzyme stabilizers are amino alcohols such as mono-, di-, triethanol- and - propanolamine and mixtures thereof, aliphatic carboxylic acids up to C ₁₂ , such as succinic acid, other dicarboxylic acids or salts of said acids. End-capped fatty acid amide alkoxylates are also suitable for this purpose. Certain organic acids used as builders are capable, as disclosed in WO 97/18287, of additionally stabilizing a contained enzyme.

Lower aliphatic alcohols, but especially polyols such as glycerol, ethylene glycol, propylene glycol or sorbitol are other frequently used enzyme stabilizers. Di-glycerol phosphate also protects against denaturation due to physical influences. Likewise, calcium and / or magnesium salts are used, such as calcium acetate or calcium formate.

Polyamide oligomers or polymeric compounds such as lignin, water-soluble vinyl copolymers or cellulose ethers, acrylic polymers and / or polyamides stabilize the enzyme preparation, inter alia, against physical influences or pH fluctuations. Polyamine N-oxide containing polymers act simultaneously as enzyme stabilizers and as dye transfer inhibitors. Other polymeric stabilizing agents are linear C ₈ -C- _I s polyoxyalkylenes. Also, alkylpolyglycosides can stabilize the enzymatic components of the agent according to the invention and, preferably, are capable of additionally increasing their performance. Crosslinked N-containing compounds preferably perform a dual function as soil release agents and as enzyme stabilizers. Hydrophobic, nonionic polymer stabilizes in particular an optionally contained cellulase.

Reducing agents and antioxidants increase the stability of the enzymes to oxidative degradation; For example, sulfur-containing reducing agents are familiar. Other examples are sodium sulfite and reducing sugars. Particular preference is given to using combinatons of stabilizers, for example of polyols, boric acid and / or borax, the combination of boric acid or borate, reducing salts and succinic acid or other dicarboxylic acids or the combination of boric acid or borate with polyols or polyamino compounds and with reducing salts. The effect of peptide-aldehyde stabilizers is favorably enhanced by the combination with boric acid and / or boric acid derivatives and polyols, and still further by the additional action of divalent cations, such as calcium ions.

Stabilized enzyme activity agents are preferred embodiments of the present invention. Particularly preferred are those with enzymes that are stabilized in several of the ways illustrated.

Cleaning agent for hard surfaces

As already mentioned, in a preferred embodiment, the subject matter of the present invention is a hard surface cleaning composition which contains a polypeptide according to the invention and / or an enzyme preparation according to the invention. The cleaning agent may contain the enzyme preparation, for example, in amounts of from 0.1 to 10.0% by weight, preferably from 0.5 to 3% by weight. The cleaning agent may optionally additionally contain biocides, in particular one of the abovementioned, but may also be biocide-free.

The hard surface cleaner is preferably a liquid, gel or pasty aqueous detergent. The hard surface cleaner may contain any of a variety of the components listed above. In a preferred embodiment, the cleaning agent contains, in addition to the polypeptide according to the invention one or more components selected from the group consisting of surfactants, builders, acids, alkalis, hydrotropes, solvents, thickeners, abrasives and other auxiliaries and additives such as dyes, perfumes, corrosion inhibitors and skin care agents , These groups will be exemplified below or have been exemplified above. The components listed below may be included in any other agent of the invention besides a hard surface cleaner. The surfactants optionally used in a hard surface cleaner are preferably selected from the group of anionic, nonionic 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 amphoteric surfactants in amounts of up to 2% by weight, based on the total composition. Likewise, cationic surfactants can also be used. In a preferred embodiment, however, the cleaning agent is free of these due to the biocidal effect emanating from cationic surfactants. The surfactants are in particular selected from the groups of the surfactants listed above.

The anionic surfactants which can be used according to the invention, in particular in hard surface cleaners, include 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 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, alkylpolyglycol ether carboxylates, fatty acid isethionates, acylaminoalkanesulfonates

(Fatty acid taurides, N-acyl taurides), fatty acid sarcosinates, ether carboxylic acids and alkyl (ether) phosphates and α-sulfo fatty acid salts, acyl glutamates,

Monoglyceride disulfates and alkyl ethers of glycerol disulfate, and finally mixtures thereof.

In the context of the present invention, fatty acids or fatty alcohols or their derivatives are-unless stated otherwise-particularly 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 10 to 18 carbon atoms, more 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 on account of their vegetable base as based on renewable raw materials for ecological reasons, but without limiting the teaching of the invention 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 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.

The anionic surfactants are preferably 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 overall composition, as well as mixtures thereof.

Suitable nonionic surfactants are, for example, C ₈ -C ₈ - alkyl alcohol polyglycol ethers, alkyl polyglycosides and nitrogen-containing surfactants or mixtures thereof, in particular the first two. C ₈ -C ₈ - Alkylalkoholpolypropylenglykol / polyethylene may by the formula

R ¹ O- (CH ₂ CH (CH ₃ ) O) _P (CH ₂ CH ₂ O) ₆ -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 ₈ -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.

Hard surface cleaners according to the present 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 for a hard surface cleaner according to the invention are also alkylpolyglycosides (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 can assume 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 used. From an application point of view, those alkyl glycosides are preferred whose degree of oligomerization is less than 1.7, and in particular between

1.2 and 1, 6 lies. 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 in the course of the hydrogenation of technical fatty acid methyl esters or in the course of the hydrogenation of aldehydes from Roelene'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 from. 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 contained in cleaners according to the invention in amounts of from 0.1 to 6% by weight, based on the total composition.

As other nonionic surfactants, nitrogen-containing surfactants may be contained, e.g. Fatty acid polyhydroxyamides, for example glucamides, and ethoxylates of alkylamines, vicinal diols and / or carboxylic acid amides which have alkyl groups with 10 to 22 C atoms, preferably 12 to 18 C 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, alkylamido alkylamines, alkyl-substituted amino acids and acylated amino acids. Preferred amphoteric surfactants are, for example, betaines of the formula

(R ³⁾ (R ⁴⁾ (R ^δ) N ⁺ CH ₂ C00;

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 _T oC-is-alkyl-dimethylcarboxymethylbetain and C ₁₁ -C ₁₇ Alkyamidopropyl 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 ^"is an anion, in particular a halide ion, for example didecyl-dimethyl-ammonium chloride, alkyl-benzyl-didecyl-ammonium chloride and mixtures thereof, but preferably the detergent composition is free of cationic surfactants.))

When choosing suitable surfactants, care must be taken 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 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.

Furthermore, the surface-active cleaners according to the invention may contain builders, in particular those selected from the group of builders listed above. Suitable builders are preferably 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, 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 Benzolhexacarbonsäure and phosphonates and Phosphates. The agents may contain builders in amounts, based on the composition, of from 0.1 to 5% by weight.

Furthermore, agents according to the invention, and in particular a hard surface cleaner 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.

Suitable bases include alkanolamines, for example mono- or diethanolamine, and ammonium or alkali metal hydroxides, especially sodium hydroxide. The hard surface cleaner of the present invention may contain bases in amounts of up to 2.5% by weight based on the total composition.

The hard surface cleaner of the invention may further contain one or more thickeners for viscosity control. 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 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 Kemmehlether 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. The inorganic thickeners which can be used 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 from 1 to 200 nm to the particle diameter in the longitudinal direction, that is, in the direction of maximum 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 C ₂ - ₈ carboxylic acids or - hydroxycarboxylic acids, functional silanes of the type (OR) _4-0 SiR _n (R = organic radicals with 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 thickening agents mentioned so far, the hard surface cleaner 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 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 composition according to the invention, it is possible to use organic and / or inorganic thickeners in amounts of up to 2% by weight, based on the total composition. The hard surface cleaner of the invention may advantageously additionally contain 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 according to the invention as needed in particular as a hydrotrope, Viskositäts¬ 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 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 composition according to the invention decreases.

Suitable solvents are for example, saturated or unsaturated, preferably saturated, branched or unbranched Ci _-2 o-hydrocarbons, preferably C ₂ i ₅ hydrocarbons, with at least one hydroxy group and optionally one or more ether functions COC, the carbon atom chain that is interrupting oxygen atoms. Preferred solvents are, however, - if appropriate on one side with a Ci _-6 etherified alkanol - C _2-6 - alkylene glycols and poly-C _2-3 -alkylene same or on average 1 to 9 different, preferably identical, alkylene glycol 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.

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 example xylene or cumene sulfonate can be used. Further usable hydrotropes are, for example, octyl sulfate or butyl glucoside. The agent of the invention may contain up to 4% by weight of these hydrotropes in amounts of, based on the total composition. In one embodiment, the hard surface cleaner of the present 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 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 according to the invention, in particular the hard surface cleaner according to the invention, may contain one or more further auxiliaries and additives, 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, as described for example in EP 522 506. 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, further Pine (spruce) and mint and floral fragrances. Preservatives have a biocidal effect, so it is desirable to use in Cleaning agents according to the invention only low concentrations, but preferably to use no preservatives.

It is advantageous if the agent contains no complexing agents; the addition of a bleach to the hard surface cleaner of the invention is not essential.

The pH of the cleaners according to the invention is in a preferred embodiment, in particular for hard surface cleaners, preferably between 1 and 8, more preferably between 2 and 5 or between 5 and 8, in particular between 2.5 and 4.5 or between 5 , 5 and 7.5. In the case of multiphase agents, the pH of the composition is understood to mean the pH of the temporary emulsion formed after the agitation; in the case of compositions offered in multi-chambered bottles, the pH of the composition is the pH the solution obtained from the common intended dosage of the components stored in the different chambers. 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 to 5% 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. in the Detergents soluble or even acting as a solvent liquefied gases (liquefied gases) as a propellant offer the advantage of constant operating pressure and uniform distribution, because in the air, the propellant evaporates, taking a multi-hundred-fold 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, however, under liquefied gases are the - hydrocarbons propane, propene, butane, butene, isobutane (2-methylpropane), isobutene (2-methylpropene), which are obtained in oil refineries as by-products in the distillation and cracking of crude oil and in natural gas treatment during gasoline separation. Isobutylene) and mixtures thereof.

The cleaning agent particularly preferably contains propane, butane and / or isobutane as one or more blowing agents, in particular propane and butane, very preferably propane, butane and isobutane.

The invention further provides a product comprising an enzyme preparation according to the invention or a composition according to the invention, in particular a hard surface cleaner according to the invention, and a spray dispenser. The product may be both a single-chamber and a multi-chamber container, in particular a two-chamber container.

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), even pressure-building spray dispenser, pump spray dispenser and trigger spray dispenser, in particular pump spray dispenser and trigger spray dispenser with a container made of transparent polyethylene or polyethylene terephthalate. Spray dispensers are described in more detail in WO 96/04940 (Procter & Gamble) and the US patents cited therein about spray dispensers, to which reference is made in this regard and the contents of which are hereby incorporated by reference. Triggersprühspender and pump sprayer have over compressed gas tanks the advantage that no propellant must be used.

In a further preferred embodiment, however, the agent containing the enzyme preparation is not aerosolized 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. In producing these compact foams, no respirable particles are formed so that the danger of inhaling allergens is substantially eliminated.

Yet another object of the invention is a method for the hydrolysis of biofilms on hard surfaces, in which the vitality of the microbial cells is preferably not damaged. For this purpose, the enzyme preparation according to the invention or the cleaning agent according to the invention, optionally using a product according to the invention, is applied to the biofilm-coated surface and, if appropriate, after an exposure time of 1 to 30 minutes, in the case of heavy infestation also for a few hours or overnight , rinsed with clean water. The preparation or the agent may also be blurred or rubbed on the surface affected by biofilm before rinsing with a cloth, a sponge, a brush or other suitable for cleaning utensil, which improves the cleaning performance, for example in the presence of abrasives in the detergent. Finally, the surface is dried with a dry cloth. If the biofilm surface is very heavily attacked, the cleaning process can then be repeated. Oral, dental and / or dental prosthesis care products

The oral, dental and / or dental prosthesis care agents according to the invention can be present, for example, as a mouthwash, gel, liquid toothbrush lotion, stiff toothpaste, chewing gum, denture cleaner or denture adhesive cream.

For this purpose, it is necessary to incorporate the hybrid enzymes according to the invention into a suitable carrier.

As carriers, e.g. also pulverulent preparations or aqueous-alcoholic solutions serve as mouthwash 0 to 15 wt .-% ethanol, 1 to 1, 5 wt .-% flavor oils and 0.01 to 0.5 wt .-% sweeteners or as mouthwash concentrates 15 to 60 wt .-% ethanol, 0.05 to 5 wt .-% aroma oils, 0.1 to 3 wt .-% sweeteners and optionally other excipients may be included and diluted with water before use. The concentration of the components must be so high that, after dilution, the specified lower concentration limits are not undershot during use.

However, gels and more or less flowable pastes, which are expressed from flexible plastic containers or tubes and applied to the teeth with the aid of a toothbrush, can also serve as the carrier. Such products contain higher levels of humectants and binders or consistency regulators and polishing components. In addition, aromatic oils, sweeteners and water are also contained in these preparations.

As humectants, e.g. Glycerol, sorbitol, xylitol, propylene glycols, Polyethenylenglycole or mixtures of these polyols, in particular those Polyethenylenglycole be used with molecular weights of 200 to 800 (from 400 to 2000). Preferably, sorbitol is contained as humectant in an amount of 25-40 wt .-%.

As anticalculus agents and as demineralization inhibitors, condensed phosphates may be present in the form of their alkali metal salts, preferably in the form of their sodium or potassium salts. The aqueous solutions of these phosphates react alkaline due to hydrolytic effects. By addition of acid, the pH of the The oral, dental and / or dental prosthesis care agents according to the invention are adjusted to the preferred values of 7.5-9.

It is also possible to use mixtures of different condensed phosphates or else hydrated salts of the condensed phosphates. The specified amounts of 2-12% by weight, however, refer to the anhydrous salts. Preferably, the condensed phosphate is a sodium or potassium tripolyphosphate in an amount of 5-10% by weight 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 fluoro compounds are e.g. Sodium monofluorophosphate (NaaPOsF), potassium monofluorophosphate, sodium or potassium fluoride, stannous 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 ^δ 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 Pfeffemninzöl, 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 present. 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 sugars such as sucrose, maltose, lactose and fructose or synthetic sweeteners such as saccharin sodium salt, sodium cyclamate or aspartame.

In particular, alkyl and / or alkenyl (oligo) glycosides can be used as surfactants. Their preparation and use as surfactants are described, for example, in US Pat. No. 3,839,318, US Pat. No. 3,707,535, US Pat. No. 3,547,828 DE-A-19 43 689, DE-A-20 36 472 and DE -A-30 01 064 and EP-A-77 167 known. With regard to the glycoside radical, monoglycosides (x = 1) in which a pentose or hexose radical is glycosidically bound to a primary alcohol having 4 to 16 carbon atoms, as well as oligomeric glycosides having a degree of oligomerization x to 10, are suitable. The degree of oligomerization is a statistical mean, which is based on a homolog distribution typical for such technical products.

As alkyl and / or alkenyl (oligo) glycoside, an alkyl and / or alkenyl (oligo) glucoside of the formula RO (C ₆ Hi ₀ 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 1 to 3. The alkyl and / or alkenyl glycoside surfactant can be very can be used sparingly, already amounts of 0.005 to 1 wt .-% are sufficient.

In addition to the alkyl glucoside surfactants mentioned, other nonionic, ampholytic and cationic surfactants may also be present, for example: 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, kylamido 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 Ci ₂ -Ci ₈ fatty acids and adducts of 20 to 60 moles of ethylene oxide with 1 mole of glycerol or 1 mole of sorbitol.

The oral, dental and / or dental prosthesis care agents according to the invention preferably contain as solubilizer for optionally contained aroma oils addition products of 20 to 60 moles of ethylene oxide to hardened or uncured castor oil (ie to Oxystearinsäure- or ricinoleic acid triglyceride), to glycerol mono- and / or distearate or sorbitan mono- and / or distearate.

Other common additives for oral, dental and / or denture care medium are, for example Pigments, for example titanium dioxide, and / or dyes, pH stabilizers and buffer substances, for example sodium bicarbonate, sodium citrate,

Sodium benzoate, citric acid, phosphoric acid or acid salts, eg NaH ₂ PO ₄ wound healing and anti-inflammatory substances such as allantoin, urea,

Panthenol, azulene or chamomile extract further anti-calculus substances such. Organophosphonates, e.g.

Hydroxyethane diphosphonates or azacycloheptane diphosphonate

Preservatives such as e.g. Sorbic acid salts, p-hydroxybenzoic acid esters.

Plaque inhibitors, e.g. Hexachlorophene, chlorhexidine, hexetidine, triclosan,

Bromochlorophene, phenylsalicylic acid ester.

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

For prosthesis cleaners according to the invention, in particular prosthesis cleansing tablets and powders, in addition to the already mentioned ingredients for oral, dental and / or dental prosthesis care, per compounds such as peroxoborate, peroxomonosulfate or percarbonate are additionally suitable. They have the advantage that, in addition to the bleaching effect, they simultaneously have a deodorizing and / or disinfecting effect. The use of such per-compounds in ^" denture cleaners is between 0.01 and 10 wt .-%, in particular between 0.5 and 5 wt .-%.

As further ingredients are also enzymes, e.g. Proteases and carbohydrase, suitable for the breakdown of proteins and carbohydrates. The pH can be between pH 4 and pH 12, in particular between pH 5 and pH 11.

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.

As active ingredients natural and synthetic swelling substances are suitable. As natural swelling agents, besides alginates, plant gums, such as e.g. Gum arabic, tragacanth and karaya gum as well as natural rubber. In particular, alginates and synthetic bulking agents, e.g. Sodium carboxymethyl cellulose, high molecular weight ethylene oxide copolymers, salts of poly (vinyl ether-co-maleic acid) and polyacrylamides.

As adjuvants for pasty and liquid products are particularly hydrophobic bases, in particular hydrocarbons, such as white Vaseline (DAB) or paraffin oil.

EXAMPLES

Example 1: Cloning of pullulanase

Shewanella sp. was isolated from Spitsbergen at 4 ⁰ C and examined for the ability to produce cold-reactive enzymes. The psychrophilic strain Shewanella sp. is a Gram-negative proteobacterium. The closest relative is Shewanella putrefaciens (51.7%), which could be demonstrated by DNA-DNA hybridization. It grows in a temperature range between 4 ° and 3O ⁰ C (optimum 10 ⁰ C) and a pH range of pH 6-10 (optimum pH 8).

Genomic DNA was isolated from Shewanella sp. isolated by standard methods (Qiagen) and digested with the restriction endonuclease Sau3A for 6 to 8 min at 37 ° C. The digested DNA was gel electrophoresed (70V, 0.8%, 45 minutes) and the bands cut out between 6 and 10 kbp and eluted from the gel piece (Nucleospin extraction kit, Macherey-Nagel).

Using the ZAP expression vector and cloning kit (Stratagene), a gene library was prepared according to the manufacturer's instructions using the isolated DNA fragments. The ZAP vector pBK-CMV (4.5 kbp) has 12 cloning sites and encodes neomycin / kanamycin resistance. To prepare the library, the fragments were ligated into lambda ZAP vector, packaged into phage proteins and transfected with these phage E. coli XLOLR cells.

100,000 E. coli XLOLR clones were screened for pullulan-containing LB kanamycin agar plates (50 μg / ml kanamycin, 1 mM IPTG, 0.1% red pullulanan) for pullulanase activity. For this purpose, the plates were incubated for 3 days at 30 ⁰ C. Active clones showed the formation of colorless courts around the colonies.

From a selected active clone, the plasmid was isolated. A portion of the plasmid, along with the 6 kbp insert, was sequenced via primer-walking. Four primer pairs were needed to obtain the complete insert sequence. The primers used are listed in Table 1. The sequence is shown in the sequence listing. Table 1: Forward and reverse primers used in primer walking

Example 2 Temperature Dependence of the Pullulanase Enzyme Activity To determine the temperature dependence, the crude extract was incubated with 1% pullulan (pH 7) for 1 h at different temperatures. The pullulanase had activities between 4 ° C and 65 ⁰ C with a temperature optimum at 45 ⁰ C (Table 2).

Table 2: Activity (U / ml) and biofilm degradation (%) of the cloned pullulanase over time

Example 3: Biofilm degradation by the pullulanase

The ability of the recombinant enzyme to degrade microbial biofilms was examined using a model biofilm grown in microtiter plates.

The biofilm test system is composed of four representatives who are considered good

Biofilmbildner are known. Model biofilms were grown on a mixed population consisting of:

Dermacoccus nishinomiyaensis DSM No. 20448 (Actinobacterium)

Bradyrhizobium japonicum DSM No. 1982 (α-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 (g tryptone, 5 g yeast extract, 5 g NaCl per liter of 10) at 30 ⁰ C and 150 rpm for 16 h were incubated in 50 ml liquid culture. Inoculation with a single colony of one agar plate (per liter 10 g tryptone, 5 g yeast extract, 5 g NaCl, 10 g agarose).

The optical density (600 nm) of the cultures after 16 h was: Dermacoccus nishinomiyaensis 7,4 Bradyrhizobium japonicum 5,4

Xanthomonas campestris 3,3

The cultures were then combined, mixed and 50% sterile glycerol added so that the final concentration was 10% glycerol. The cultural mix was portioned and frozen at -20 ⁰ 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. The plates were incubated for 16 h at different temperatures with 1 ml of the enzyme extract. Depending on the temperature, the enzyme was able to break down up to 28% of the biofilm, with the strongest activity being observed at 30 ^° C. At higher temperatures, little or no biofilm degradation was measurable (Table 2).

Claims

claims

A polynucleotide selected from the group consisting of: a) polynucleotide having a nucleic acid sequence according to SEQ ID NO: 1 or having a nucleic acid sequence from position 70 to position 4323 according to SEQ ID NO: 1, b) polynucleotide encoding a polypeptide having an amino acid sequence according to SEQ ID NO: 2 or for a polypeptide having an amino acid sequence from position 24 to position 1440 according to SEQ ID NO: 2, c) naturally occurring or artificially produced mutants or polymorphic forms or alleles of a polynucleotide according to (a) or (b), which d) polynucleotides complementary to a polynucleotide according to (a), (b) or (c), e) under stringent conditions with a polynucleotide according to (a), up to 50 point mutations with respect to a polynucleotide according to (a) or (b); (b), (c) or (d) hybridizing polynucleotides, f) polynucleotides having a sequence homology or identity of at least 60% with respect to a polynucleotide according to (a), (b), ( c) or (d), g) polynucleotides consisting of at least 25 consecutive nucleic acids of a polynucleotide according to (a), (b), (c), (d) or (e), h) polynucleotides with deletions and / or insertions and or inversions of up to 50 nucleotides compared to a polynucleotide according to (a), (b), (c), (d) or (e), i) polynucleotides comprising at least one of the polynucleotides mentioned under (a) to (h) ,

2. polynucleotide according to claim 1, characterized in that it codes for a hydrolase.

3. Polynucleotide according to claim 2, characterized in that it is a glycosylase in the hydrolase.

4. Polynucleotide according to claim 3, characterized in that the glycosylase is a pullulanase.

5. Polynucleotide according to one of claims 1 to 4, characterized in that the encoded polypeptide is capable of cleaving the bonds between α-1, 6-gknüpften D-glucose units.

6. A method for producing and / or identifying a polynucleotide according to any one of claims 1 to 5, characterized in that the polynucleotide is chemically synthesized, isolated from a Genbank by means of a probe or obtained by polymerase chain reaction.

7. A vector comprising a nucleic acid according to any one of claims 1 to 5.

8. A host cell comprising a vector according to claim 7.

9. polypeptide selected from the group consisting of: a) polypeptide having an amino acid sequence according to SEQ ID NO: 2 or having an amino acid sequence from position 24 to position 1440 according to SEQ ID NO: 2, b) naturally occurring mutants, polymorphic forms or alleles of one Polypeptides according to (a) having up to 50 point mutations with respect to a polypeptide according to (a), c) polypeptides having a sequence homology or identity of at least 60% with respect to a polypeptide according to (a) or (b) , d) polypeptides encoded by a polynucleotide according to claim 1, e) polypeptides consisting of at least 8 consecutive amino acids of a polypeptide according to (a) or (b), f) polypeptides having insertions and / or deletions and / or inversions of bis to 60 amino acids with respect to a polypeptide according to (a), (b) or (c), g) polypeptides comprising at least one of the polypeptides mentioned under (a) to (f).

10. Polypeptide according to claim 9, characterized in that it is a hydrolase.

11. Polypeptide according to claim 10, characterized in that it is a glycosylase in the hydrolase.

12. Polypeptide according to claim 11, characterized in that the hydrolase is a pullulanase.

13. Polypeptide according to one of claims 9 to 12, characterized in that the polypeptide is able to cleave bonds between α-1, 6-gknüpften D-glucose units.

14. An enzyme preparation comprising a polypeptide according to any one of claims 9 to 13.

15. An enzyme preparation according to claim 14, characterized in that it additionally contains enzymes selected from polysaccharidases, proteases and nucleases.

16. A composition selected from the group consisting of sterilizing, disinfecting, washing, dishwashing and cleaning agents, characterized in that it contains a polypeptide according to any one of claims 9 to 13 or an enzyme preparation according to claim 14 or 15.

17. The composition according to claim 16, characterized in that the polypeptide and / or the enzyme preparation are contained in amounts of up to 10 wt .-% in the preparation.

18. The composition of claim 16 or 17, characterized in that it contains at least one further component selected from the group consisting of surfactants, Buildem, acids, alkalis, hydrotropes, solvents, thickeners, bleaching agents, dyes, perfumes, corrosion inhibitors, sequestering agents , Electrolytes, optical brighteners, grayness inhibitors, silver corrosion inhibitors,

Dye transfer inhibitors, foam inhibitors, abrasives, UV Absorbents, solvents, abrasives, antistatic agents, pearlescing agents and skin protection agents.

19. A composition according to any one of claims 16 to 18, characterized in that it is a solid, gel-like, pasty or liquid composition.

20. A product comprising a polypeptide according to any one of claims 9 to 13, an enzyme preparation according to claim 14 or 15 or a composition according to any one of claims 16 to 19 and a spray dispenser for dosing the enzyme preparation or the cleaning agent as an aerosol and / or as a foam.

21. A method for hydrolysis of biofilms on surfaces or for the removal of soil residues on surfaces by incubation with a polypeptide according to any one of claims 9 to 13, an enzyme preparation according to claim 14 or 15 or a composition according to any one of claims 16 to 19, optionally using A product according to claim 20, characterized in that the vitality of the microbial cells is not permanently damaged.

22. A method for the hydrolysis of biofilms on surfaces, characterized in that the polypeptide, the enzyme preparation or the cleaning agent according to one of the preceding claims, optionally using a product according to the invention according to claim 20, is applied to the biofilm-coated surface, optionally with a Cloth, a sponge, a brush or other suitable for cleaning utensil on the biofilm-infested surface is blurred or rubbed and, optionally after a contact time of 1 to 30 minutes, a few hours or overnight, rinsed with clear water, whereupon wipe the surface with a dry cloth.

23. Use of polypeptides according to any one of claims 9 to 13, an enzyme preparation according to claim 14 or 15 or a composition according to any one of claims 16 to 19 for the destruction and / or removal of biofilms and / or for the removal of debris on surfaces.

24. Use of polypeptides according to any one of claims 9 to 13, an enzyme preparation according to claim 14 or 15 or a composition according to any one of claims 16 to 19 for the degradation of pullulan.

25. Use of polypeptides according to any one of claims 9 to 13, an enzyme preparation according to claim 14 or 15 or a composition according to any one of claims 16 to 19 for the hydrolytic cleavage of α-1, 4 and / or α-1, 6 bonds between D-monosaccharides.

26. Use of polypeptides according to any one of claims 9 to 13, an enzyme preparation according to claim 14 or 15 or a composition according to any one of claims 16 to 19 for the purification of textiles.

27. Product of an enzyme preparation or a cleaning agent according to one of the preceding claims and a multi-chamber bottle.

28. Cosmetic and / or pharmaceutical preparation containing polypeptides according to one of claims 9 to 13 or an enzyme preparation according to claim 14 or 15.

29. A cosmetic and / or pharmaceutical preparation according to claim 28, characterized in that it is in the preparation to oral, dental or denture dentifrice.

30. Use of polypeptides according to any one of claims 9 to 13 or an enzyme preparation according to claim 14 or 15 for the preparation of a cosmetic preparation.

31. Use of polypeptides according to any one of claims 9 to 13 or an enzyme preparation according to claim 14 or 15 for the manufacture of a medicament for the prevention or degradation of dental plaque.

32. Bacterium Shewanella sp. 40-3 according to DSM 16509.

33. Hydrolases obtainable from Shewanella sp. 40-3 according to DSM 16509.

34. Use of Shewanella sp. 40-3 for degradation of biofilms or for removal of debris from surfaces.

35. Use of Shewanella sp. 40-3 for the extraction of pullulan.

36. Use of Shewanella sp. 40-3 for the hydrolytic cleavage of α-1, 4- and / or α-1, 6 bonds between D-monosaccharides.

37. A method for isolating and / or identifying hydrolases from psychrophilic bacteria, comprising the steps of: a) isolating genomic DNA from a psychrophilic bacterium, b) digesting the genomic DNA by means of suitable restriction endonucleases, c) incorporating the DNA bands in one size d) preparing a gene bank by transfecting a host with the transfection vector, e) screening the gene bank for cells containing enzymes with desired activity, f) optionally isolating the plasmid with the clone having the desired activity , and g) optionally sequencing the isolated clone.