WO2008089860A2

WO2008089860A2 - Methods for the enzymatic production of hydrogen peroxide

Info

Publication number: WO2008089860A2
Application number: PCT/EP2007/062837
Authority: WO
Inventors: Thomas Weber; Timothy O'connell; Nina Hoven; Ralf Weidenhaupt; Astrid Spitz; Karl-Heinz Maurer
Original assignee: Henkel Ag & Co. Kgaa
Priority date: 2007-01-26
Filing date: 2007-11-27
Publication date: 2008-07-31
Also published as: WO2008089860A3; DE102007005053A1

Abstract

The invention relates to methods for the enzymatic production of hydrogen peroxide, wherein the use of choline or choline derivatives as the substrates for the oxidase enzymes used in these methods can be avoided. The invention also relates to novel oxidase enzymes that are suitable for the use in these methods and that are not restricted to choline or choline derivatives as the preferred substrates. Finally, the invention relates to uses of said novel oxidase enzymes and to agents containing said novel oxidase enzymes.

Description

Process for the enzymatic production of hydrogen peroxide

The present invention relates to processes for the enzymatic production of hydrogen peroxide, in which it is possible to dispense with the use of choline or choline derivatives as substrate for the oxidase enzymes used in these processes. The invention further relates to novel oxidase enzymes which are suitable for use in these methods and which are distinguished by the ability not to be limited to choline or choline derivatives as a substrate and to be able to oxidize alcohols of the R-CH 2 -OH type. Furthermore, the invention relates to uses of these novel oxidase enzymes as well as agents containing these novel oxidase enzymes.

Choline oxidases have been known in the art as important oxidase enzyme representatives (Ikuta, S., Imamura, S., Misaki, H., and Horiuti, Y. 1977. Purification and characterization of choline oxidase from Arthrobacter globiformis J Biochem (Tokyo) 82: 1741-1749, Deshnium, P., Los, DA, Hayashi, H., Mustardy, L., and Murata, N. 1995. Transformation of Synechococcus with a gene for choline oxidase enhances tolerance to salt stress Plant Mol Biol 29: 897-907.). It is also known that oxidases (for example alcohol oxidases or amino acid oxidases) can be used together with their substrates for hydrogen peroxide generation for bleaching and dye transfer inhibition in detergents or for enzymatic hair coloring and bleaching in cosmetics. Oxidases may therefore be part of an enzymatic bleaching system.

In the publication by Gadda et al. (Arch. Biochem., Biophys., 421 (1), 149-158, 2004). The authors describe a choline oxidase from the "cloning, sequence analysis, and purification of choline oxidase from Arthrobacter globiformis: a bacterial enzyme involved in osmotic stress tolerance" Bacterium Arthrobacter globiformis (Database Accession Number AAP68832 of NCBI (National Center for Biotechnology Information) - Protein database; UniProtKB / TrEMBL entry Q7X2H8), which in the present application is indicated as Sequence 2 (SEQ ID NO: 2) and in particular for sequence comparisons is used.

For conventional chemical bleaching, inorganic, alkaline hydrogen peroxide sources such as percarbonate or perborate are used in combination with bleach booster (TAED). The bleaching component hydrogen peroxide is formed by spontaneous decomposition of the addition compound and thus leads quickly but quickly to high concentrations. A gentle bleaching is not possible.

For hair dyeing, bleaching is conventionally carried out for leveling shades of gray before dyeing with hydrogen peroxide and ammonia solution. The short-term high Hydrogen peroxide concentrations in combination with the alkaline pH lead to a significant hair damage.

In the German patent application 102 60 930.6-41 Amelderin it is stated that and how can be isolated from the bacteria Arthrobacter nicotianae and Arthrobacter aurescens choline oxidases, which can be used advantageously in bleaching systems. The Arthrobacter nicotianae strain KC2, which contains the choline oxidase according to SEQ ID NO: 3, is deposited with DSMZ GmbH (German Collection of Microorganisms and Cell Cultures GmbH) in 38124 Braunschweig, Inhoffenstraße 7 B, Germany (DSMZ-ID 96-878, Deposit number DSM 11234). The disclosure of this application is therefore expressly incorporated into the present application.

However, a choline oxidase preferably converts choline as a substrate. In general, however, the availability of enzymes with different substrate specificity is desirable. Depending on the question, the preferred substrate can be used in a single process.

From the prior art, publications are concerned with the substrate binding and the catalytic mechanism of choline oxidase enzymes. For example, Gadda et al. (Arch. Biochem., Biophys. 430 (2), 264-73, 2004. "The trimethylammonium headgroup of choline is a major determinant for substrate binding and specificity in choline oxidase") the importance of the positively charged trimethylammonium group of choline for the Substrate binding and specificity of enzymatic catalysis disclosed. In this context, Ghanem et al. (Biochemistry 44 (3), 893-904, 2005. "On the catalytic role of the conserved active site resident His466 of choline oxidase") the contribution of the histidine residue at position 466 of the choline oxidase used to the catalytic mechanism to see how the substrate specificity of the disclosed choline oxidase enzymes can be changed in favor of substrates other than choline or choline derivatives.

From the publication by Lountos, George T. (2005, "Structural and mechanistic insights from the toluene-4-monooxygenase catalytic effector protein, NAD (P) H oxidase and choline oxidase", ISBN 0-542-43394 -X) it is known that replacement of the amino acid glutamic acid at position 312 of the choline oxidase used, which is involved in substrate binding, to alanine or aspartate significantly reduces the activity of choline oxidase for the conversion of choline.With further amino acid residues involved in substrate binding W61 However, there is no disclosure from the cited publication that a change in the amino acid at said positions results in a change in the substrate specificity of the oxidase activity in favor of other substrates other than W331, H351, F357, V464, Y465 and H466 Choline distinguish leads. International patent application WO 2007045398 describes choline oxidase enzymes with optimized specificity towards the substrates choline, bis (2-hydroxyethyl) dimethylammonium chloride (dimethyldiethanolammonium chloride, DMDEA) and triethanolamine (TEA) and tris (2-hydroxyethyl) methylammonium chloride (MTEA). These enzymes are variants of the choline oxidase KC2 from the bacterium Arthrobacter nicotianae, which have mutations in molecular regions or domains, based on the choline oxidase KC2, which are selected from the substrate binding domain and the FAD binding domain. Preferably, one or more mutations are selected at amino acid sequence positions selected from positions 4, 21, 62, 69, 79, 116, 128, 166, 215, 243, 249, 260, 286, 321, 348, 349, 351, 393, 394, 530 and 531, wherein the numbering of the sequence positions is related to the sequence 3 (SEQ ID NO: 3) given in the present application. However, there is no suggestion that a targeted reduction of the activity of the enzymes towards the substrate choline can be used to increase the activity towards substrates which differ from the substrates mentioned in this application.

The processes known from the prior art for the enzymatic production of hydrogen peroxide, in particular those which contain an enzymatic catalytic step by choline oxidases, which are known from the prior art, have the disadvantage that an enzymatic oxidation of choline, which is used as substrate for The choline oxidase serves to cause severe odor problems due to trimethylamine. There is thus a need to be able to dispense with choline as the substrate of choline oxidase in such a process and thus to be able to use such enzymes in these processes, which are capable of reacting a substrate other than choline.

Furthermore, choline is a biomolecule present in biological systems. The production of oxidase in cells in which choline is also present causes oxidative stress for the enzyme-producing cells under aerobic conditions due to the resulting hydrogen peroxide. This impairs enzyme production, for example, by obtaining insoluble or oxidatively damaged protein, poor yield, poor reproduction of the fermentation processes, increased catalase production, etc. Thus, there is a great need for oxidases, their substrates, or only a small proportion in the production cells occurrence.

The present invention is therefore based on the object to provide a process for the enzymatic production of hydrogen peroxide, in which can be dispensed with the use of choline or choline derivatives as a substrate for the oxidase enzymes, which are used in this method. Another object of the present invention is to provide just such oxidase enzymes, which are characterized by the ability to oxidize alcohols of the type R-CH2-OH and are not limited to choline or choline derivatives as a substrate and in particular for the oxidation of a Substrates, which is different from choline, are better capable than for the oxidation of choline.

This object is achieved according to the teaching of claim 1 by the application and provision of oxidases which are obtained by altering the amino acid sequence of the choline oxidase KC2 from the bacterium Arthrobacter nicotianae or partial sequences thereof (SEQ ID NO: 3 and SEQ ID NO: 1) and, because of their alteration, are capable of oxidizing substrates other than choline and choline derivatives, and are more capable of oxidizing a substrate other than choline, rather than oxidizing choline. In this regard, SEQ ID NO: 1 represents a variant of this choline oxidase truncated by six amino acids in comparison with SEQ ID NO: 3 (FIG. 1). The Arthrobacter nicotianae strain KC2, which contains the choline oxidase according to SEQ ID NO: 3, is deposited with DSMZ GmbH (German Collection of Microorganisms and Cell Cultures GmbH) in 38124 Braunschweig, Inhoffenstraße 7 B, Germany (DSMZ-ID 96-878, Deposit number DSM 11234). According to the invention, the oxidases preferably have changes in those amino acid residues which influence the structure of the substrate binding domain of the oxidase. Oxidases according to the invention can therefore be used advantageously in processes in which, for certain reasons, no choline can or should be used. To clarify the fact that the enzymes according to the invention are obtained by altering a choline oxidase, but are advantageously not used for the oxidation of choline or choline derivatives as substrate, the enzymes according to the invention are not in the present application as choline oxidases, but as oxidase enzymes or oxidases.

An object of the present invention thus provide methods for the enzymatic production of hydrogen peroxide with an oxidase, which are characterized in that the amino acid sequence of the oxidase includes an amino acid sequence which is at least 96% identical to SEQ ID NO: 1, with reaction of a substrate, which is different from choline. However, it is also possible to use oxidases whose amino acid sequence includes an amino acid sequence which is at least 97%, 98%, 99%, 99.5% and further 100% identical to SEQ ID NO: 1.

In one embodiment of the invention, the method of the invention is characterized in that the substrate other than choline is one having the structure R-CH 2 -OH, wherein R is 1-20 carbon atoms, 0-5 nitrogen atoms, 0 -5 Oxygen atoms, 0-2 sulfur atoms, 0-2 phosphorus atoms and 0-10 halogen atoms. The substrate may be in saturated or unsaturated form.

In another embodiment, the substrate other than choline is one having the structure R-CH 2 -OH, wherein R is selected from the group consisting of -phenyl, -benzyl, -CH 2-SO 2 -OH, CH2-PO (OH) 2, -PO (OH) 2, -CH2-tert-butyl, -CH2-C (Me2, OH). Particularly preferred embodiments of the invention are characterized in that the substrate other than choline is 2-phenylethanol, 3-methyl-1,3-butanediol, isethionic acid (2-hydroxyethanesulfonic acid) or 3-benzyl alcohol.

In the process according to the invention, generally all substrates whose volume is less than 2 × 10 ^-8 m ³ can be used, since the binding pocket of the oxidase used in the process has this volume. Thus, a further embodiment of the invention is characterized in that the substrate has a volume which is smaller than 2x 10 ^{~ 8} m ³ .

For the purposes of the present application, an enzyme is to be understood as meaning a protein which has a specific biocatalytic function. For the purposes of the present application, oxidases are understood as meaning enzymes which transfer the electrons released during the oxidation of a substrate to oxygen (O ₂ ). The oxygen serves as an electron acceptor and is reduced to water (H ₂ O) or hydrogen peroxide (H ₂ O ₂ ). The activity of an oxidase may require a non-proteinaceous cofactor which may be free or covalently or non-covalently bound to the oxidase. For example, these may be flavin adenine dinucleotide "FAD" or redox-active metal ions.

For the purposes of the present application, a protein is a polypeptide composed largely of linear structure composed of the natural amino acids and generally assuming a three-dimensional structure in order to perform its function. A peptide consists of amino acids that are covalently linked to each other via peptide bonds. The term polypeptide clarifies in this regard the fact that this peptide chain usually consists of many amino acids, which are connected to each other via peptide bonds. Amino acids may be in an L and a D configuration, with the amino acids that make up proteins in the L configuration. They are called proteinogenic amino acids. In the present application, the proteinogenic, naturally occurring L-amino acids are designated by the internationally used 1- and 3-letter codes. Numerous proteins are formed as so-called pre-proteins, ie together with a signal peptide. By this is meant the N-terminal part of the protein, the function of which is usually to ensure the discharge of the protein formed from the producing cell into the periplasm or the surrounding medium and / or its correct folding. Subsequently, the signal peptide under natural conditions by a signal peptidase from the remaining protein cleaved so that this exerts its actual catalytic activity without the initially present N-terminal amino acids. Pro-proteins are inactive precursors of proteins. Their signal sequence precursors are referred to as pre-pro proteins. For technical applications, due to their enzymatic activity, the mature, ie mature, peptides, ie the enzymes processed after their preparation, are preferred over the preproteins. The proteins may be modified by the cells producing them after production of the polypeptide chain, for example, by attachment of sugar molecules, formylations, aminations, etc. Such modifications are referred to as post-translational modifications. These post-translational modifications may or may not have an effect on the function of the protein.

By comparison with known enzymes, which are deposited, for example, in generally accessible databases, the enzymatic activity of a considered enzyme can be deduced from the amino acid or nucleotide sequence. This can be qualitatively or quantitatively modified by other regions of the protein that are not involved in the actual reaction. This could, for example, relate to enzyme stability, activity, reaction conditions or substrate specificity.

Such a comparison is accomplished by associating similar sequences in the nucleotide or amino acid sequences of the proteins of interest. This is called homologization. A tabular assignment of the respective positions is referred to as alignment. In the analysis of nucleotide sequences, in turn, both complementary strands and all three possible reading frames are to be taken into account; as well as the degeneracy of the genetic code and the organism-specific codon usage. Meanwhile, alignments are created using computer programs, such as the algorithms FASTA or BLAST; this procedure is described, for example, by D.J. Lipman and W.R. Pearson (1985) in Science, Vol. 227, pp. 1435-1441.

A summary of all matching positions in the compared sequences is called a consensus sequence.

Such a comparison also allows a statement about the similarity or homology of the compared sequences to each other. This is expressed in percent identity, that is, the proportion of identical nucleotides or amino acid residues at the same positions. A broader concept of homology includes the conserved amino acid substitutions in this value. It then speaks of percent similarity. Such statements can be made about whole proteins or genes or only over individual areas. Homologous regions of different proteins are defined by matches in amino acid sequence. These can also be identified by identical function. It goes as far as complete identities in the smallest areas, so-called boxes, which contain only a few amino acids and usually perform essential functions for the overall activity. The functions of the homologous regions are to be understood as the smallest partial functions of the function carried out by the entire protein, such as, for example, the formation of individual hydrogen bonds for the complexation of a substrate or transition complex.

Another object of the invention are formed oxidases, which are characterized in that one or more amino acids at positions 55, 306, 325, 345, 351, 458, 459 and 460 are changed with respect to SEQ ID NO: 1, wherein the amino acid to one of the positions 55, 325, 345, 351, 458, 459 and 460 is selected from the group of alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, Proline, serine, threonine, tryptophan, tyrosine and valine and is different from that present in SEQ ID NO: 1 at the appropriate position and the amino acid at position 306 is selected from the group of arginine, asparagine, cysteine, glutamine , Glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine and valine.

Surprisingly, it has been found that altering the amino acid in one or more of these positions, while diminishing but unexpectedly increasing the enzyme's ability to oxidize choline as a substrate, increases the ability of the enzyme to oxidize additional new substrates. Thus, such a change in the amino acid causes a change in the specific activity of the enzyme. Particularly surprising and noteworthy is the possibility of substrate change away from the positively charged trimethylammonium group of the choline towards uncharged or negatively charged groups of the new substrates.

Further, altering an amino acid in sequence neighborhood, preferably in immediate sequence neighborhood, to the amino acids at positions 55, 306, 325, 345, 351, 458, 459 and 460 relative to SEQ ID NO: 1 may also result in a change in substrate specificity. since such a change can lead to shifts in the amino acids involved in substrate binding. Such oxidases are therefore included in the present invention. According to the invention, sequence proximity means a position which is within a window of 50 amino acid positions, more preferably of 40, 30, 20, 15, 10, 8, 6, 5, 4, 2 and especially preferably of an amino acid position, starting from specifically named amino acid in both directions, ie both towards the N-terminus and towards the C-terminus of the amino acid sequence. Basically, the amino acid substitutions proposed by the present application are not limited to being the only substitutions in which the subject oxidase is different from the parent molecule. It is generally known from the prior art that advantageous properties of individual point mutations can complement one another. An oxidase which has already been optimized with regard to certain properties, for example with respect to its stability towards surfactants or other components, can be further developed according to the invention via the substitutions presented here. Thus, embodiments of the present invention include all oxidases which, in addition to other exchanges, also have exchanges according to the invention.

Also included are oxidases obtainable from the abovementioned oxidases according to the invention by derivatization, fragmentation, inversion mutation, deletion mutation or insertion mutation, wherein it is of course imperative that the corresponding mutation position or combination of mutation positions in the resulting oxidases be retained.

Fragments are understood as meaning all proteins or peptides which are smaller than natural proteins and, for example, can be obtained synthetically. Due to their amino acid sequences, they can be assigned to the relevant complete proteins. For example, they may adopt the same structures or perform proteolytic or partial activities, such as the complexation of a substrate. Fragments and deletion variants of starting proteins are in principle similar; while fragments tend to be smaller fragments, the deletion mutants tend to lack only short regions, and thus only individual subfunctions.

For the purposes of the present application, chimeras or hybrid proteins are to be understood as meaning those proteins whose sequence comprises the sequences or partial sequences of at least two starting proteins. The source proteins may be derived from different or from the same organism. Chimeric or hybrid proteins may be obtained, for example, by recombinant mutagenesis. For example, the purpose of such recombination may be to induce or modify a particular enzymatic function using the fused protein portion. For the purposes of the present invention, it is irrelevant whether such a chimeric protein consists of a single polypeptide chain or several subunits on which different functions can be distributed.

By proteins obtained by insertion mutation are meant those variants obtained by inserting a protein fragment into the starting sequences. They are due to their principle similarity to the chimeric proteins. They differ of those only in the size ratio of the unaltered portion of protein to the size of the entire protein. In such insertionsmutierten proteins, the proportion of foreign protein is lower than in chimeric proteins.

Inversion mutagenesis, ie a partial sequence reversal, can be regarded as a special form of both the deletion and the insertion. The same applies to a new grouping of different parts of the molecule which deviates from the original amino acid sequence. It can be regarded as a deletion variant, as an insertion variant, as well as a shuffling variant of the original protein.

For the purposes of the present application, derivatives are understood as meaning those proteins whose pure amino acid chain has been chemically modified. Such derivatizations can be carried out, for example, biologically in connection with protein biosynthesis by the host cell. For this molecular biological methods can be used. However, they can also be carried out chemically, for example by the chemical transformation of a side chain of an amino acid or by covalent binding of another compound to the protein. Such a compound may, for example, also be other proteins which are bound, for example via bifunctional chemical compounds, to proteins according to the invention. Such modifications may, for example, affect the substrate specificity or binding strength to the substrate or cause a temporary blockage of the enzymatic activity when the coupled substance is an inhibitor. This can be useful, for example, for the period of storage. Also, derivatization is understood to mean covalent attachment to a macromolecular carrier, as well as noncovalent inclusion in suitable macromolecular cage structures.

Proteins can also be grouped into groups of immunologically related proteins by reaction with an antiserum or antibody. The members of a group are characterized by having the same antigenic determinant recognized by an antibody.

For the purposes of the present invention, all enzymes, proteins, fragments and derivatives, unless they need to be addressed explicitly as such, are summarized under the generic term proteins.

The oxidases according to the invention with altered substrate specificity advantageously have a high specific hydrogen peroxide formation rate. The pH profile of the enzymes according to the invention is compatible with the required pH for industrial use and for use in typical products such as detergents and cleaners and hair dyes.

In order to be used advantageously in the processes described above, the oxidases according to the invention are able to oxidize substrates which differ from choline. Another object of the invention, therefore, are oxidases, which are characterized in that they are better able to oxidize a substrate, which is different from choline, as for the oxidation of choline. Also included are oxidases which are more capable of oxidizing a substrate different from choline than the unaltered starting enzyme.

In a further embodiment of the invention, the oxidase is characterized in that the substrate other than choline is one having the structure R-CH 2 -OH, wherein R is 1-20 carbon atoms, 0-5 nitrogen atoms, 0 Contains -5 oxygen atoms, 0-2 sulfur atoms, 0-2 phosphorus atoms and 0-10 halogen atoms.

In a preferred embodiment the oxidase being characterized in that it is in the substrate, which is different from choline to a ₂ is -OH having the structure R-CH, wherein R is selected from the group consisting of phenyl, - benzyl, -CH ₂ -SO ₂ -OH, -CH ₂ - PO (OH) ₂ , -PO (OH) ₂ , -CH ₂ -tert-butyl, -CH2-C (Me ₂ , OH). In particularly preferred embodiments of the invention, the oxidases are characterized in that the substrate other than choline is 2-phenylethanol, 3-methyl-1,3-butanediol, isethionic acid or benzyl alcohol.

The oxidases of the present invention are capable of accepting substrates whose volume is less than 2 × 10 ^-8 m ³ in their substrate binding pocket since the binding pocket of an oxidase used in the method has this volume. Thus, further embodiments of the oxidases according to the invention are characterized in that the substrate has a volume which is smaller than 2 × 10 ^-8 m ³ .

In a further embodiment of the invention, the oxidases are characterized in that the amino acid at position 306 is increasingly preferably arginine, glutamine, asparagine, aspartate or glycine, the numbering of the sequence positions being based on SEQ ID NO: 1.

However, oxidases according to the invention are expressly not limited to this embodiment, but may contain at position 306 the amino acid arginine, asparagine, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, Tyrosine or valine and positions 55, 325, 345, 351, 458, 459 and 460 the amino acids alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine and valine, where the numbering is related to SEQ ID NO: 1. These amino acids can be present at the positions mentioned in any combination. Furthermore, several of the abovementioned positions or else only one of the abovementioned positions can have the corresponding amino acid. As already explained above, embodiments of the present invention include all oxidases which, in addition to other exchanges, also have exchanges according to the invention.

The oxidases according to the invention have a pH optimum preferably in the almost neutral to weakly alkaline range of about pH 6 to pH 10, particularly preferably pH 7 to pH 9. The activity of such enzymes is usually expressed in units E (or "Unit", U), one unit corresponding to that amount of enzyme which generates 1 μmol of hydrogen peroxide (H ₂ O ₂ ) at a fixed pH and temperature in 1 minute . the temperature optimum of the oxidase according to the invention is approximately in the range of 20 to 6O ⁰ C, particularly at about 4O ⁰ C.

Since the subject oxidases of the present application are better or at least equivalent to the oxidation of a substrate other than choline, their activity to react a substrate other than choline is higher or at least equivalent their activity by reacting choline as a substrate. Thus, another subject of the invention is oxidases for which the quotient formed from the specific activity of the oxidase using a substrate other than choline divided by the specific activity of the oxidase using a substrate which is choline Value between 1 and 100,000, preferably a value between 1 and 1000, more preferably a value between 1 and 100 and particularly preferably has a value between 1 and 10.

Further objects of the invention form nucleic acid molecules which code for an oxidase according to the invention as well as vectors containing such a nucleic acid.

For the purposes of the present application, nucleic acids are understood to mean the molecules which are naturally constructed from nucleotides and serve as information carriers, which code for the linear sequence of amino acids in proteins or enzymes. They can be present as a single strand, as a single strand that is complementary to this single strand, or as a double strand. As the naturally more durable information carrier, the nucleic acid DNA is preferred for molecular biology work. In contrast, for the realization of the invention in a natural environment, such as in an expressing cell, an RNA is formed, why RNA molecules essential to the invention are also embodiments of the present invention.

For DNA, consider the sequences of both complementary strands in all three possible reading frames. Further, it should be appreciated that different codon triplets may encode the same amino acids so that a particular amino acid sequence may be derived from a plurality of different and possibly low identity nucleotide sequences, which is referred to as the degeneracy of the genetic code. In addition, various organisms have differences in the use of these codons. For these reasons, both amino acid sequences and nucleotide sequences must be included in the consideration of the scope. Therefore, all nucleotide sequences are included in the invention which can encode any of the oxidases described above. The skilled person is able to determine these nucleotide sequences beyond doubt, because despite the degeneracy of the genetic code individual codons defined amino acids are assigned. Therefore, one of ordinary skill in the art, on the basis of one example, may be considered only as an exemplary coding for a particular amino acid sequence.

The information unit corresponding to a protein is also referred to as gene within the meaning of the present application.

The present invention involves the production of recombinant proteins. According to the invention, these are to be understood as meaning all genetic engineering or microbiological processes which are based on the genes for the proteins of interest being introduced into a suitable host cell for the production and being transcribed and translated by it. Suitably, the introduction of the relevant genes via vectors, in particular expression vectors; but also those that cause the gene of interest in the host cell to be inserted into an already existing genetic element, such as the chromosome or other vectors. The functional unit of gene and promoter and any other genetic elements is referred to as expression cassette according to the invention. However, it does not necessarily have to exist as a physical entity.

A person skilled in the art can use well-known methods such as chemical synthesis or the polymerase chain reaction (PCR) in combination with molecular biological and / or proteinchemical standard methods, using known DNA and / or amino acid sequences, the corresponding nucleic acids to complete genes manufacture.

Changes in the nucleotide sequence, as can be brought about, for example, by molecular biological methods known per se, are referred to as mutations. Depending on Type of change is known, for example, deletion, insertion or substitution mutations or those in which different genes or parts of genes are fused or recombined with each other; these are gene mutations. The associated organisms are called mutants. The proteins derived from mutant nucleic acids are called variants. Thus, for example, deletion, insertion substitution mutations or fusions lead to deletion, insertion-substitution-mutated or fusion genes and, at the protein level, to corresponding deletion, insertion or substitution variants or fusion proteins.

For the purposes of the present invention, vectors are understood as consisting of nucleic acids which contain a gene of interest as a characteristic nucleic acid region. They are able to establish this in a species or a cell line over several generations or cell divisions as a stable genetic element. Vectors, especially when used in bacteria, are special plasmids, ie circular genetic elements. One differentiates in the genetic engineering on the one hand between those vectors which serve the storage and thus to a certain extent also the genetic engineering, the so-called cloning vectors, and on the other hand those which fulfill the function of realizing the gene of interest in the host cell, that is, the expression of the protein. These vectors are referred to as expression vectors.

In the context of the present invention, the nucleic acid is suitably cloned into a vector. The molecular-biological dimension of the invention thus consists in vectors with the genes for the corresponding proteins. These may include, for example, those derived from bacterial plasmids, viruses or bacteriophages, or predominantly synthetic vectors or plasmids with elements of various origins. With the other genetic elements present in each case, vectors are able to establish themselves as stable units in the relevant host cells over several generations. For the purposes of the invention, it is irrelevant whether they establish themselves as extrachomosomal units or integrate them into a chromosome. Which of the numerous systems known from the prior art is chosen depends on the individual case. Decisive factors may be, for example, the achievable copy number, the selection systems available, in particular antibiotic resistances, or the cultivability of the host cells capable of accepting the vectors.

The vectors form suitable starting points for molecular biological and biochemical investigations of the relevant gene or protein and for further developments according to the invention and ultimately for the amplification and production of proteins according to the invention. They represent insofar embodiments of the present invention, as the Sequences of the nucleic acid regions according to the invention in each case lie within the homology regions specified above.

A further subject of the invention thus represents vectors which contain at least one nucleic acid molecule which codes for an oxidase according to the invention, which are characterized in that the vector is a cloning vector. These are suitable in addition to the storage, the biological amplification or the selection of the gene of interest for the characterization of the gene in question, such as the creation of a restriction map or sequencing. Cloning vectors are also preferred embodiments of the present invention because they are a transportable and storable form of the claimed DNA. They are also preferred starting points for molecular biological techniques that are not bound to cells, such as the polymerase chain reaction (PCR).

A further subject of the invention furthermore provides vectors which contain at least one nucleic acid molecule which codes for an oxidase according to the invention, which are characterized in that the vector is an expression vector. Expression vectors are chemically similar to the cloning vectors, but differ in those partial sequences which enable them to replicate in the host cells or host organisms optimized for the production of proteins and to express the gene contained therein. Preferred embodiments are expression vectors which themselves carry the genetic elements necessary for expression. The expression is influenced, for example, by promoters which regulate the transcription of the gene. Thus, expression may be by the natural promoter originally located upstream of this gene, but also by genetic engineering, both by a host cell promoter provided on the expression vector and by a modified or completely different promoter from another organism or host cell.

Preferred embodiments are those expression vectors which are regulatable via changes in culture conditions or addition of certain compounds, such as cell density or specific factors. Expression vectors allow the associated protein to be produced heterologously, that is in a cell or host cell other than that from which it can naturally be obtained. The cells may well belong to different organisms or come from different organisms. Also, homologous protein recovery from a gene cell naturally expressing the gene via an appropriate vector is within the scope of the present invention. This may have the advantage that natural translational-related modification reactions on the resulting protein are performed exactly as they would naturally occur. A further embodiment represents expression systems in which additional genes, for example those which are provided on other vectors, influence the production of proteins according to the invention. These may be modifying gene products or those which are to be purified together with the protein according to the invention, for example in order to influence its enzymatic function. These may be, for example, other proteins or enzymes, inhibitors or elements that influence the interaction with various substrates.

Alternative embodiments of the present invention may also be cell-free expression systems in which protein biosynthesis is understood in vitro. Such expression systems are also established in the art.

A further subject of the invention is a host cell which contains an oxidase according to the invention or a fragment thereof or which can be excited to produce it, preferably using an expression vector. The in vivo synthesis of an enzyme according to the invention, ie by living cells, requires the transfer of the associated gene into a host cell, the so-called transformation thereof. In principle, all cells, that is to say prokaryotic or eukaryotic cells, are suitable as host cells. Preference is given to those host cells which can be genetically advantageously handled, for example, the transformation with the expression vector and its stable establishment, for example, unicellular fungi or bacteria. In addition, preferred host cells are characterized by good microbiological and biotechnological handling. This concerns, for example, easy culturing, high growth rates, low demands on fermentation media and good production and secretion rates for foreign proteins. Frequently, the optimal expression systems for the individual case must be determined experimentally from the abundance of different systems available according to the prior art. Each protein of the invention can be obtained in this way from a variety of host cells. Also, those host cells are preferred, which are characterized in that they have been obtained after transformation with one of the vectors described above. These may, for example, be cloning vectors which have been introduced for storage and / or modification, for example into any bacterial strain or another host cell according to the invention. Such steps are common in the storage and further development of related genetic elements. Since the relevant genetic elements can be directly transferred from these host cells into subsequent host cells suitable for expression, the preceding transformation products are also realizations of the subject matter of the invention.

Preferred embodiments represent such host cells which, due to genetic regulatory elements provided for example on the expression vector but can also be present in these cells from the outset, are regulatable in their activity. For example, by controlled addition of chemical compounds that serve as activators, by changing the culture conditions or when reaching a specific cell density, these can be excited for expression. This allows for economical production of the proteins of interest.

Preferred host cells are prokaryotic or bacterial cells. As a rule, bacteria are distinguished from eukaryotes by shorter generation times and lower demands on culturing conditions. As a result, cost-effective methods for obtaining proteins according to the invention can be established. In Gram-negative bacteria, such as Escherichia coli (E. coli), a large number of proteins are secreted into the periplasmic space, ie into the compartment between the two membranes enclosing the cells. This can be advantageous for special applications. Gram-positive bacteria, such as, for example, Bacilli or Actinomycetes or other representatives of the Actinomycetales, have no outer membrane, so that secreted proteins are readily released into the nutrient medium surrounding the cells, from which, according to a further preferred embodiment, the expressed proteins according to the invention can be directly purified.

Preferred dimensions of the host cell is therefore characterized in that it secretes the protein of the invention or a fragment or derivative thereof into the surrounding medium.

In a further preferred embodiment, the host cell according to the invention is characterized in that it is a bacterium, in particular selected from the group of the genera of Escherichia, Bacillus and Arthrobacter, Streptomyces and Pseudomonas. The host cell is particularly preferably a bacterium which is selected from the group of Escherichia coli, Bacillus licheniformis, Bacillus amyloliquefaciens, Bacillus subtilis, Bacillus alcalophilus and Arthrobacter oxidans, Streptomyces lividans and Streptomyces coelicolor.

The host cells may be altered in their culture conditions requirements, have different or additional selection markers, or express other or additional proteins. In particular, these may be those host cells which, in addition to the protein produced according to the invention, also express further, in particular economically interesting, proteins.

The host cell may also be a eukaryotic cell, which is characterized in that it has a cell nucleus. In contrast to prokaryotic cells, eukaryotic cells are capable of post-translationally modifying the protein formed. Examples thereof are fungi such as Actinomycetes or yeasts such as Saccharomyces or Kluyveromyces. This can For example, then be particularly advantageous if the proteins are to undergo specific modifications in connection with their synthesis, which allow such systems. These include, for example, the binding of low molecular weight compounds such as membrane anchors or oligosaccharides.

The host cells of the process according to the invention are cultivated and fermented in a manner known per se, for example in discontinuous or continuous systems. In the first case, a suitable nutrient medium is inoculated with the host cells and the product is harvested from the medium after an experimentally determined period of time. Continuous fermentations are characterized by achieving a flow equilibrium in which over a relatively long period of time cells partly die off but also regrow and at the same time product can be removed from the medium.

Fermentation processes are known per se from the prior art and represent the actual large-scale production step, usually followed by a suitable purification method of the product produced, for example the recombinant protein. All fermentation processes which are based on one of the above-described processes for the preparation of the recombinant proteins represent correspondingly preferred embodiments of this subject matter of the invention.

In this case, the optimum conditions for the production processes used, for the host cells and / or the proteins to be produced must be experimentally determined on the basis of the previously optimized culture conditions of the relevant strains according to the knowledge of the skilled person, for example regarding fermentation volume, media composition, oxygen supply or stirrer speed.

Fermentation processes, which are characterized in that the fermentation is carried out via a feed strategy, are also contemplated. Here, the media components consumed by the ongoing cultivation are fed; One also speaks of a feeding strategy. As a result, considerable increases in both the cell density and in the dry biomass and / or especially the activity of the protein of interest can be achieved.

Similarly, the fermentation can also be designed so that unwanted metabolic products are filtered out or neutralized by the addition of buffer or matching counterions.

The produced protein can be harvested subsequently from the fermentation medium. This fermentation process is compared to the product preparation from the dry mass preferred, but requires the provision of suitable secretion markers and transport systems. Without secretion may be necessary to purify the protein from the cell mass, and various methods are known, such as precipitation, for example by ammonium sulfate and / or ethanol, or the chromatographic purification, if necessary, to homogeneity. However, the majority of the described technical methods should manage with an enriched, stabilized preparation.

All of the elements already described above can be combined to form methods for producing proteins according to the invention. It is conceivable for each protein of the invention a variety of possible combinations of process steps. The optimal procedure has to be determined experimentally for each specific case.

By expression or cloning the oxidases of the invention can be made available in the amount required for industrial use.

A further subject of the invention is the use of an oxidase according to the invention as a hydrogen peroxide generating agent. A preferred embodiment provides the use of the oxidase as hydrogen peroxide in situ, i. Locally generating agent. The fact that the formation of hydrogen peroxide takes place at the place where the oxidase is located, it can be used advantageously in the methods and means of the invention. Further preferred embodiments of a use according to the invention are the use for bleaching, for color transfer inhibition and for disinfection.

A further subject of the invention are agents containing an oxidase according to the invention. The oxidases according to the invention or the oxidases usable according to the invention can advantageously be used in personal care products, shampoos, hair care products, oxidation dyes, oral, dental or denture care products, cosmetics, detergents, cleaners, rinse-offs , Detergents, dishwashing detergents, dishwashing detergents, disinfectants, signal reagents or agents for the bleaching or disinfecting treatment of filter media, textiles, furs, paper, skins or leather.

Preferred in the invention is an agent containing a bleach system which is capable of producing hydrogen peroxide under conditions of use of the agent. The bleaching system preferably consists of an oxidase according to the invention and a substrate for the oxidase. The substrate preferably differs from choline as described in detail above.

An oxidase according to the invention can be used both in agents for large consumers or technical users as well as in products for the private consumer, all established in the prior art detergents and cleaning agents also embodiments of the present invention. These include, for example, concentrates and undiluted agents for use on a commercial scale, in the washing machine or in hand washing or cleaning. Likewise, for example, laundry detergents for textiles, carpets, or natural fibers, for which according to the present invention, the term detergent is used. Furthermore, these include, for example, dishwashing detergents for dishwashers or manual dishwashing detergents or cleaners for hard surfaces such as metal, glass, porcelain, ceramics, tiles, stone, painted surfaces, plastics, wood or leather; for such according to the present invention, the term cleaning agent is used.

Embodiments of the present invention include all established and / or all appropriate dosage forms. These include, for example, solid, powdery, liquid, gelatinous or pasty agents, which may optionally also consist of several phases and may be present in compressed or uncompressed form. Another subject of the invention are therefore agents which are characterized in that they are present as one-component systems. A further subject of the invention is constituted by means characterized in that it is divided into several components. The dosage forms according to the invention also include extrudates, granules, tablets or pouches, which may be present both in large packages and in portions.

In a preferred embodiment of the invention, the agent is characterized in that it is present as a free-flowing powder having a bulk density of 300 g / l to 1200 g / l, in particular 500 g / l to 900 g / l.

In a further preferred embodiment of the invention, the agent is characterized in that it is in pasty or liquid form, in particular in the form of a non-aqueous liquid detergent or a non-aqueous paste or in the form of an aqueous liquid detergent or a water-containing paste.

The composition according to the invention, in particular washing or bleaching agent, may be packaged in an air-impermeable container from which it is released shortly before use or during the washing process, in particular the oxidase and / or the substrate for this enzyme may be present at room temperature or in the absence be enveloped by water for the enzyme and / or its substrate impermeable substance which becomes permeable under conditions of use of the agent for the enzyme and / or its substrate. The agent is thus characterized in that the oxidase and / or its substrate is coated with a substance which is impermeable to the oxidase and / or its substrate at room temperature or in the absence of water. The oxidases according to the invention are combined in inventive compositions, for example with one or more of the following ingredients: nonionic, anionic and / or cationic surfactants, (optionally further) bleaching agents, bleach activators, bleach catalysts, builders and / or cobuilders, solvents, thickeners, sequestering agents, electrolyte , optical brighteners, grayness inhibitors, corrosion inhibitors, in particular silver protectants, soil release agents, color transfer (or transfer) inhibitors, foam inhibitors, abrasives, dyes, perfumes, antimicrobial agents, UV protectants, enzymes such as proteases, amylases, lipases, Cellulases, hemicellulases or oxidases, stabilizers, especially enzyme stabilizers, and other components known in the art.

The ingredients to be selected as well as the conditions under which the agent is used, such as temperature, pH, ionic strength, redox ratios or mechanical influences, should be optimized for the particular cleaning problem. Thus, conventional temperatures for detergents and cleaners in the range from 1 O ⁰ C for manual agents over 40 ⁰ C and 60 ⁰ C up to 95 ° for mechanical means or in technical applications. Since the temperature is usually infinitely adjustable in modern washing machines and dishwashers, all intermediate stages of the temperature are included. Preferably, the ingredients of the respective agents are coordinated. Synergies in terms of cleaning performance are preferred.

The agent according to the invention, in particular washing or bleaching agent, preferably contains in addition to the bleaching system

From 5% by weight to 70% by weight, in particular from 5% by weight to 30% by weight, of surfactants,

10% by weight to 65% by weight, in particular 12% by weight to 60% by weight of water-soluble or water-dispersible inorganic builder material,

0.5% by weight to 10% by weight, in particular 1% by weight to 8% by weight, of water-soluble organic builders,

0.01 to 15% by weight of solid inorganic and / or organic acids or acid salts,

0.01 to 5% by weight complexing agent for heavy metals,

0.01 to 5% by weight of grayness inhibitor,

0.01 to 5% by weight of color transfer inhibitor and

0.01 to 5% by weight foam inhibitor

Optionally, the agent may further comprise optical brighteners as well as one or more further enzymes. The optical brighteners are preferably used from 0.01 to 5% by weight, the other enzymes are preferably used from 0.01 to 5 wt .-%. The agent according to the invention, in particular washing or bleaching agent, preferably has an oxidase activity of 0.01 units / g to 20 000 units / g. Preferred is an oxidase activity of at least 5000 units per 75 g of detergent or bleach.

The compositions according to the invention or the compositions prepared by the process according to the invention described above contain washing or cleaning substances from the group of builders, surfactants, polymers, bleaches, bleach activators, bleach catalysts, enzymes, glass corrosion inhibitors, corrosion inhibitors, disintegration aids, fragrances and perfume carriers , These preferred ingredients will be described in more detail below.

builders

The builders include, in particular, the zeolites, silicates, carbonates, organic cobuilders and, where there are no ecological prejudices against their use, also the phosphates.

The finely crystalline, synthetic and bound water-containing zeolite used 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) ), by the formula

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

can be described. The zeolite can be used both as a builder in a granular compound and for a kind of "powdering" of a granular mixture, preferably a mixture to be compressed, whereby usually both ways of incorporating the zeolite into the premix are used an average particle size of less than 10 microns (volume distribution, measuring method: Coulter Counter) and preferably contain 18 to 22 wt .-%, in particular 20 to 22 wt .-% of bound water.

With preferred crystalline layered silicates of general formula NaMSi _x O _{2x + I} ^■ y H ₂ O are used, wherein M is sodium or hydrogen, x is a number from 1, 9 to 22, preferably from 1: 9 to 4, wherein particularly preferred Values for x are 2, 3 or 4, and y is a number from 0 to 33, preferably from 0 to 20. The crystalline layered silicates of the formula NaMSi _x O _{2x + 1} ^■ y H ₂ O are sold for example by Clariant GmbH (Germany) under the trade name Na-SKS. Examples of these silicates are Na-SKS-1 (Na ₂ Si ₂₂ O ₄₅ ^.H ₂ O, Kenyaite), Na-SKS-2 (Na ₂ Si ₁₄ O ₂₉ ^■ x H ₂ O, magadiite), Na-SKS-3 (Na ₂ Si ₈ O ₁₇ ^■ x H ₂ O) or Na-SKS-4 (Na ₂ Si ₄ O ₉ ^.H ₂ O, makatite).

Particularly suitable for the purposes of the present invention are crystalline layer silicates with the formula NaMSi _x O _{2x + 1} ^■ y H ₂ O, in which x stands for 2 h. In particular, both .beta.- and sodium disilicates δ- Na ₂ Si ₂ O ₅ ^■ y H ₂ O, and further in particular Na-SKS-5 (Oc-Na ₂ Si ₂ O _5), Na-SKS-7 (.beta.-Na ₂ Si ₂ O _5, natrosilite), Na-SKS-9 (NaHSi ₂ O ₅ ^■ H ₂ O), Na-SKS-10 (NaHSi ₂ O ₅ ^■ 3 H ₂ O, kanemite), Na SKS (11 t-Na ₂ Si ₂ O ₅ ) and Na-SKS-13 (NaHSi ₂ O ₅ ), but especially Na-SKS-6 (5-Na ₂ Si ₂ O ₅ ) are preferred.

Washing or cleaning composition preferably contain a weight proportion of crystalline layered silicate of the formula NaMSi _x O _{2x + 1} ^■ y H ₂ O from 0.1 to 20 wt .-%, preferably from 0.2 to 15 wt .-% and in particular of 0.4 to 10 wt .-%, each based on the total weight of these agents.

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 preferably delayed release 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 understood to mean that the silicates do not yield sharp X-ray reflections typical of crystalline substances in X-ray diffraction experiments, but at most one or more maxima of the scattered X-rays having a width of several degrees of diffraction angle , cause.

Alternatively or in combination with the abovementioned amorphous sodium silicates, X-ray-amorphous silicates are used whose silicate particles give washed-out or even sharp diffraction maxima in electron diffraction experiments. This is to be interpreted as meaning that the products have microcrystalline regions of the size of ten to a few hundred nm, with values of up to max. 50 nm and in particular up to max. 20 nm are preferred. Such X-ray amorphous silicates also have a dissolution delay compared to conventional water glasses. Particularly preferred are compacted / compacted amorphous silicates, compounded amorphous silicates and overdried X-ray amorphous silicates.

In the context of the present invention, it is preferred that these silicate (s), preferably alkali metal silicates, particularly preferably crystalline or amorphous alkali metal disilicates, be present in detergents or cleaners in amounts of from 3 to 60% by weight, preferably from 8 to 50 Wt .-% and in particular from 20 to 40% by weight, based in each case on the weight of the washing or cleaning agent.

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 a particular preference for pentasodium or pentapotassium triphosphate (sodium or potassium tripolyphosphate), have the 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 (HPO ₃ ) _n and orthophosphoric H ₃ PO _{4 in} addition to higher 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.

Technically particularly important phosphates are the pentasodium triphosphate, Na ₅ P ₃ O ₁₀ (sodium tripolyphosphate) and the corresponding potassium salt pentapotassium triphosphate, K ₅ P ₃ O ₁₀ (potassium tripolyphosphate). The sodium potassium tripolyphosphates are also preferably used according to the invention.

If phosphates are used as detergents or cleaning agents in the context of the present application, preferred agents comprise these phosphate (s), preferably alkali metal phosphate (s), more preferably pentasodium or pentapotassium triphosphate (sodium or pentasodium) Potassium tripolyphosphate), in amounts of from 5 to 80% by weight, preferably from 15 to 75% by weight, in particular from 20 to 70% by weight, based in each case on the weight of the washing or cleaning agent.

Further builders are the alkali carriers. Suitable alkali carriers are, for example, alkali metal hydroxides, alkali metal carbonates, alkali metal hydrogencarbonates, alkali metal sesquicarbonates, the alkali silicates mentioned, alkali metal silicates, and mixtures of the abovementioned substances, preference being given to using alkali metal carbonates, in particular sodium carbonate, sodium bicarbonate or sodium sesquicarbonate for the purposes of this invention. Particularly preferred is a builder system comprising a mixture of tripolyphosphate and sodium carbonate. Because of their low chemical compatibility with the other ingredients of detergents or cleaners compared with other builders, the alkali metal hydroxides are preferably only in small amounts, preferably in amounts below 10 wt .-%, preferably below 6 wt .-%, more preferably below 4 wt .-% and in particular below 2 wt .-%, each based on the total weight of the washing or Detergent used. Particularly preferred are agents which, based on their total weight, contain less than 0.5% by weight and in particular no alkali metal hydroxides.

Particularly preferred is the use of carbonate (s) and / or bicarbonate (s), preferably alkali metal carbonate (s), more preferably sodium carbonate, in amounts of 2 to 50 wt .-%, preferably from 5 to 40 wt .-% and in particular from 7.5 to 30 wt .-%, each based on the weight of the washing or cleaning agent. Particular preference is given to compositions which, based on the weight of the washing or cleaning agent, contain less than 20% by weight, preferably less than 17% by weight, preferably less than 13% by weight and in particular less than 9% by weight of carbonate ( e) and / or bicarbonate (s), preferably alkali metal carbonate (s), particularly preferably sodium carbonate.

Particularly suitable organic co-builders are polycarboxylates / polycarboxylic acids, polymeric polycarboxylates, aspartic acid, polyacetals, dextrins, other organic cobuilders (see below) and phosphonates. These classes of substances are described below.

Useful organic builders are, for example, the polycarboxylic acids which can be used in the form of the free acid and / or 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 is not objectionable for ecological reasons, and mixtures of these. In addition to their builder effect, the free acids also typically have the property of an acidifying component and thus also serve to set a lower and milder pH of detergents or cleaners. In particular, citric acid, succinic acid, glutaric acid, adipic acid, gluconic acid and any desired mixtures of these can be mentioned here.

Further suitable builders are polymeric polycarboxylates, for example the alkali metal salts of polyacrylic acid or of 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 against polystyrenesulfonic acids measured molar masses are generally much 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 detergents or cleaners to (co) polymeric polycarboxylates is preferably 0.5 to 20 wt .-%, in particular 3 to 10 wt .-%.

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 contain as monomers salts of acrylic acid and maleic acid and vinyl alcohol or vinyl alcohol derivatives or as monomers salts of acrylic acid and 2-alkylallylsulfonic acid and sugar derivatives ,

Further preferred copolymers are those which have as their monomers 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.

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 common measure of the reducing action of a polysaccharide compared to dextrose, which has a DE of 100 , is. 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.

Oxydisuccinates and other derivatives of disuccinates, preferably ethylenediamine disuccinate, are other suitable co-builders. Ethylenediamine-N, N ^'- disuccinate (EDDS) is preferably used in form of its sodium or magnesium salts. Also preferred in this context are glycerol disuccinates and glycerol trisuccinates. Suitable amounts are in zeolithhaltigen and / or silicate-containing formulations at 3 to 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.

In addition, all compounds capable of forming complexes with alkaline earth ions can be used as builders.

The group of surfactants includes nonionic, anionic, cationic and amphoteric surfactants.

As nonionic surfactants, it is possible to use all nonionic surfactants known to the person skilled in the art. Suitable nonionic surfactants are, for example, alkyl glycosides of the general formula RO (G) _x in which R is a primary straight-chain or methyl-branched, in particular 2-methyl-branched aliphatic radical having 8 to 22, preferably 12 to 18 carbon atoms and G is the symbol that is for a glycose unit with 5 or 6 C atoms, preferably for glucose. The degree of oligomerization x, which indicates the distribution of monoglycosides and oligoglycosides, is an arbitrary number between 1 and 10; preferably x is 1, 2 to 1, 4.

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.

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 amount of these nonionic surfactants is preferably not more than that of the ethoxylated fatty alcohols, especially not more than half thereof.

Further suitable surfactants are polyhydroxy fatty acid amides of the formula

wherein R is 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] is a linear or branched polyhydroxyalkyl radical having 3 to 10 carbon atoms and 3 to 10 hydroxyl groups. 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

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 or an oxyalkyl radical having 1 to 8 carbon atoms, wherein Ci_ ₄ alkyl or phenyl radicals are preferred and [Z] is a linear polyhydroxyalkyl radical whose alkyl chain is substituted with at least two hydroxyl groups, or alkoxylated, preferably ethoxylated or propoxylated derivatives of this radical.

[Z] is preferably obtained by reductive amination of a reduced 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 by reaction with fatty acid methyl esters in the presence of an alkoxide as catalyst.

Low-foaming nonionic surfactants are used as preferred surfactants. With particular preference, washing or cleaning agents, in particular automatic dishwashing detergents, contain nonionic surfactants from the group of the alkoxylated alcohols. The nonionic surfactants used are preferably alkoxylated, advantageously ethoxylated, in particular primary, alcohols having preferably 8 to 18 carbon atoms and an average of 1 to 12 moles of ethylene oxide (EO) per mole of alcohol, in which the alcohol residue can be linear or preferably methyl-branched in the 2-position or linear and methyl-branched radicals in the mixture can contain, 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 of coconut, palm, tallow or oleyl alcohol, and on average 2 to 8 moles of EO per mole of alcohol are preferred. Preferred ethoxylated alcohols include, for example, C _t2 - _U - alcohols with 3 EO or 4 EO, C ₉ n-alcohol with 7 EO, C-ms 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 C ₁₂ -i ₄ -alcohol with 3 EO and C ₁₂ -i ₈ -alcohol with 5 EO. The stated degrees of ethoxylation represent statistical averages, which may correspond to a particular product of 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.

With particular preference, therefore, ethoxylated nonionic surfactant selected from C ₆ - o- ₂ monohydroxy alkanols or C ₆ - ₂ o-alkyl phenols or _C16 - ₂ o-fatty alcohols and more than 12 mol, preferably more than 15 mol and in particular more than 20 mol Ethylene oxide per mole of alcohol were used. A particularly preferred nonionic surfactant is selected from a straight chain fatty alcohol having 16 to 20 carbon atoms (C _{_16-2} alcohol), preferably a C ₁₈ alcohol obtained and at least 12 mole, preferably at least 15 mol and in particular at least 20 moles of ethylene oxide. Of these, the so-called "narrow ranks ethoxylates" are particularly preferred. With particular preference further combinations of one or more Taigfettalkoholen be used with 20 to 30 EO and silicone defoamers.

Particular preference is given to nonionic surfactants which have a melting point above room temperature. Nonionic surfactant (s) (e) with a melting point above 2O ⁰ C, preferably above 25 ⁰ C, more preferably between 25 and 6O ⁰ C and in particular from 26.6 to 43.3 ⁰ C, is / are particularly preferred ,

Suitable nonionic surfactants which have melting or softening points in the temperature range mentioned are, for example, low-foaming nonionic surfactants which may be solid or highly viscous at room temperature. If nonionic surfactants are used which are highly viscous at room temperature, it is preferred that they have a viscosity above 20 Pas, preferably above 35 Pas and in particular above 40 Pas. Also, nonionic surfactants having waxy consistency at room temperature are preferred depending on their purpose.

Nonionic surfactants from the group of alkoxylated alcohols, more preferably from the group of mixed alkoxylated alcohols and in particular from the group of EO-AO-EO-Niotenside, are also used with particular preference.

The nonionic surfactant solid at room temperature preferably has propylene oxide units in the molecule. Preferably, such PO units make up to 25 wt .-%, more preferably up to 20 wt .-% and in particular up to 15 wt .-% of the total molecular weight of the nonionic surfactant from. Particularly preferred nonionic surfactants are ethoxylated monohydroxyalkanols or alkylphenols which additionally have polyoxyethylene-polyoxypropylene block copolymer units. The alcohol or alkylphenol content of such nonionic surfactant molecules preferably makes up more than 30% by weight, more preferably more than 50% by weight and in particular more than 70% by weight, of the total molecular weight of such nonionic surfactants. Preferred agents are characterized in that they contain ethoxylated and propoxylated nonionic surfactants in which the propylene oxide units in the molecule up to 25 wt .-%, preferably up to 20 wt .-% and in particular up to 15 wt .-% of the total molecular weight of the nonionic Make up surfactants.

Preferably used surfactants come from the groups of alkoxylated nonionic surfactants, in particular the ethoxylated primary alcohols and mixtures of these surfactants with structurally complicated surfactants such as polyoxypropylene / polyoxyethylene / polyoxypropylene ((PO / EO / PO) surfactants). Such (PO / EO / PO) nonionic surfactants are also characterized by good foam control. More particularly preferred nonionic surfactants having melting points above room temperature contain from 40 to 70% of a polyoxypropylene / polyoxyethylene / polyoxypropylene block polymer blend containing 75% by weight of a reverse block copolymer of polyoxyethylene and polyoxypropylene with 17 moles of ethylene oxide and 44 moles of propylene oxide and 25% by weight. % of a block copolymer of polyoxyethylene and polyoxypropylene initiated with trimethylolpropane and containing 24 moles of ethylene oxide and 99 moles of propylene oxide per mole of trimethylolpropane.

In the context of the present invention, particularly preferred nonionic surfactants have been low foaming nonionic surfactants which have alternating ethylene oxide and alkylene oxide units. Among these, in turn, surfactants with EO-AO-EO-AO blocks are preferred, wherein in each case one to ten EO or AO groups are bonded to each other before a block of the other groups follows. Here are nichionic surfactants of the general formula

Ri-O- (CH ₂ -CH ₂ -O) - (CH ₂ -CHO) - (CH ₂ -CH ₂ -O) - (CH ₂ -CHO) -H

R2 R3

preferably, R ¹ is a straight-chain or branched, saturated or mono- or polyunsaturated C ₆ - ₂₄ represents alkyl or alkenyl; each group R ² or R ^{3 is} independently selected from -CH ₃ , -CH ₂ CH ₃ , -CH ₂ CH ₂ -CH ₃ , CH (CH ₃ ) ₂ and the indices w, x, y, z independently stand for integers from 1 to 6.

The preferred nonionic surfactants of the above formula can be prepared by known methods from the corresponding alcohols R ¹ -OH and ethylene or alkylene oxide. The radical R ¹ in the above formula may vary depending on the origin of the alcohol. If native sources are used, the radical R ^{1 has} an even number of carbon atoms and is usually unbranched, the linear radicals being selected from alcohols of natural origin having 12 to 18 C atoms, for example from coconut, palm, tallow or Oleyl alcohol, are preferred. Alcohols which are accessible from synthetic sources are, for example, the Guerbet alcohols or methyl-branched or linear and methyl-branched radicals in the 2-position, as usually present in oxo alcohol radicals. Irrespective of the type of alcohol used to prepare the nonionic surfactants contained in the compositions, preference is given to nonionic surfactants in which R ¹ in the above formula is an alkyl radical having 6 to 24, preferably 8 to 20, particularly preferably 9 to 15 and in particular 9 to 11 Carbon atoms.

As the alkylene oxide unit which is contained in the preferred nonionic surfactants in alternation with the ethylene oxide unit, in particular butylene oxide is considered in addition to propylene oxide. But also other alkylene oxides in which R ² or R ^{3 are} independently selected from -CH ₂ CH ₂ -CH ₃ or CH (CH ₃ ) ₂ are suitable. Preference is given to using nonionic surfactants of the above formula in which R ² or R ^{3 is} a radical -CH ₃ , w and x independently of one another for values of 3 or 4 and y and z independently of one another are values of 1 or 2.

In summary, particular preference is given to nonionic surfactants which have a

having 1 to 4 ethylene oxide units followed by 1 to 4 propylene oxide units followed by 1 to 4 ethylene oxide units followed by 1 to 4 propylene oxide units. These surfactants have the required low viscosity in aqueous solution and can be used according to the invention with particular preference.

Surfactants of the general formula

R ^{1 is} -CH (OH) CH ₂ O- (AO) _w - (AO) _x - (A "O) _y - (A" O) _z -R ² in which

R ¹ and R ² independently of one another represent a straight-chain or branched, saturated or mono- or polyunsaturated C ₂ - ₄ o-alkyl or alkenyl radical; A, A ', A "and A'" independently represent a radical from the group -CH ₂ CH ₂ , -CH ₂ CH ₂ -CH ₂ , -CH ₂ -CH (CH ₃ ), -CH ₂ -CH ₂ - CH ₂ -CH ₂ , -CH ₂ -CH (CHs) -CH ₂ -, -CH ₂ -CH (CH ₂ -CH ₃ ); and w, x, y and z are values between 0.5 and 90, where x, y and / or z can also be 0 are preferred according to the invention.

Preference is given in particular to those end-capped poly (oxyalkylated) nonionic surfactants which are of the formula

R ¹ O [CH ₂ CH ₂ O] _x CH ₂ CH (OH) R ²

in addition to a radical R ¹ , which is linear or branched, saturated or unsaturated, aliphatic or aromatic hydrocarbon radicals having 2 to 30 carbon atoms, preferably having 4 to 22 carbon atoms, further having a linear or branched, saturated or unsaturated, aliphatic or aromatic hydrocarbon radical R2 Have from 1 to 30 carbon atoms, wherein x stands for values between 1 and 90, preferably for values between 40 and 80 and in particular for values between 40 and 60.

Particularly preferred are surfactants of the formula

R ¹ O [CH ₂ CH (CH ₃ ) O] _x [CH ₂ CH ₂ O] _y CH ₂ CH (OH) R ² ,

in which R ^{1 is} a linear or branched aliphatic hydrocarbon radical having 4 to 18 carbon atoms or mixtures thereof, R ² denotes a linear or branched hydrocarbon radical having 2 to 26 carbon atoms or mixtures thereof and x for values between 0.5 and 1, 5 and y is a value of at least 15. Particular preference is furthermore given to those end-capped poly (oxyalkylated) nonionic surfactants of the formula

R ¹ O [CH ₂ CH ₂ O] _x [CH ₂ CH (R ³ ) O] _y CH ₂ CH (OH) R ² ,

in which R ¹ and R ² independently of one another are a linear or branched, saturated or mono- or polyunsaturated hydrocarbon radical having 2 to 26 carbon atoms, R ^{3 is} independently selected from -CH ₃ , -CH ₂ CH ₃ , -CH ₂ CH ₂ -CH ₃ , CH (CH ₃ ) ₂ , but preferably represents -CH ₃ , and x and y independently of one another are values between 1 and 32, wherein nonionic surfactants with R ³ = -CH ₃ and values for x of 15 to 32 and y of 0.5 and 1.5 are most preferred.

Other preferred nonionic surfactants are the end-capped poly (oxyalkylated) nonionic surfactants of the formula

R ¹ O [CH ₂ CH (R ³ P] _x [CH ₂ J _k CH (OH) [CH ₂ ] PR ² ,

in which R ¹ and R ^{2 are} linear or branched, saturated or unsaturated, aliphatic or aromatic hydrocarbon radicals having 1 to 30 carbon atoms, R ^{3 is} H or a methyl, ethyl, n-propyl, iso-propyl, n- Butyl, 2-butyl or 2-methyl-2-butyl radical, x are values between 1 and 30, k and j are values between 1 and 12, preferably between 1 and 5. If the value x> 2, each R ³ in the above formula

R ¹ O [CH ₂ CH (R ³ ) O] _x [CH ₂ ] _k CH (OH) [CH ₂ ] _J OR ^{2 be} different. R ¹ and R ² are preferably linear or branched, saturated or unsaturated, aliphatic or aromatic hydrocarbon radicals having 6 to 22 carbon atoms, with radicals having 8 to 18 carbon atoms being particularly preferred. For the radical R ³ , H, -CH ₃ or -CH ₂ CH _{3 are} particularly preferred. Particularly preferred values for x are in the range from 1 to 20, in particular from 6 to 15.

As described above, each R ³ in the above formula may be different if x> 2. As a result, the alkylene oxide unit in the square bracket can be varied. For example, when x is 3, the radical R ³ can be selected to form ethylene oxide (R ³ = H) or propylene oxide (R ³ = CH ₃ ) units which may be joined in any order, for example (EO) ( PO) (EO), (EO) (EO) (PO), (EO) (EO) (EO), (PO) (EO) (PO), (PO) (PO) (EO) and (PO) ( PO) (PO). The value 3 for x has been selected here by way of example and may well be greater, the range of variation increasing with increasing x values and including, for example, a large number (EO) groups combined with a small number (PO) groups, or vice versa , Particularly preferred end-capped poly (oxyalkylated) alcohols of the above formula have values of k = 1 and j = 1, so that the above formula zu

R ¹ O [CH ₂ CH (R ³ ) O] _X CH ₂ CH (OH) CH ₂ OR ²

simplified. In the latter formula, R ¹ , R ² and R ^{3 are} as defined above and x is from 1 to 30, preferably from 1 to 20 and in particular from 6 to 18. Particularly preferred are surfactants in which the radicals R ¹ and R ^{2 has} 9 to 14 C atoms, R ^{3 is} H and x assumes values of 6 to 15.

The stated C chain lengths and degrees of ethoxylation or degrees of alkoxylation of the abovementioned nonionic surfactants represent statistical mean values which, for a specific product, may be an integer or a fractional number. Due to the manufacturing process, commercial products of the formulas mentioned are usually not made of an individual representative, but of mixtures, which may result in mean values for the C chain lengths as well as for the degrees of ethoxylation or degrees of alkoxylation and subsequently broken numbers.

Of course, the aforementioned nonionic surfactants can be used not only as individual substances, but also as surfactant mixtures of two, three, four or more surfactants. Mixtures of surfactants are not mixtures of nonionic surfactants which fall in their entirety under one of the abovementioned general formulas, but rather mixtures which contain two, three, four or more nonionic surfactants which can be described by different general formulas ,

As anionic surfactants, for example, those of the sulfonate type and sulfates are used. Suitable surfactants of the sulfonate type are preferably C ₉ .i ₃ -alkylbenzenesulfonates, olefinsulfonates, ie mixtures of alkene and hydroxyalkanesulfonates and also disulfonates, as are known for example from C 1 _{_2-18} monoolefins with terminal or internal double bond by sulfonation with gaseous sulfur trioxide and subsequent alkaline or acidic hydrolysis of the sulfonation products into consideration. Also suitable are alkanesulfonates which are obtained from C ₁₂ -is alkanes, for example by sulfochlorination or sulfoxidation with subsequent hydrolysis or neutralization. Similarly, the esters of α-sulfo fatty acids (ester sulfonates), for example, the α-sulfonated methyl esters of hydrogenated coconut, palm kernel or Taigfettsäuren are suitable.

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 in the preparation by esterification of a monoglycerol with 1 to 3 mol fatty acid or obtained in the transesterification of triglycerides with 0.3 to 2 moles of glycerol. Preferred sulfated fatty acid glycerol esters are the sulfonation products of saturated fatty acids containing 6 to 22 carbon atoms, for example caproic acid, caprylic acid, capric acid, myristic acid, lauric acid, palmitic acid, stearic acid or behenic acid.

Suitable alk (en) ylsulfates are the alkali metal salts and, in particular, the sodium salts of the sulfuric acid semiesters of C ₁₂ -C ₁₈ fatty alcohols, for example coconut fatty alcohol, tallow fatty alcohol, lauryl, myristyl, cetyl or stearyl alcohol or the C ₁₀ -C ₂ o- 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 C ₁₂ -C ₁₆ alkyl sulfates and C ₁₂ -C ₁₅ alkyl sulfates and C ₁₄ -C ₁₅ - alkyl sulfates are preferred. 2,3-alkyl sulfates, which can be obtained as commercial products from Shell Oil Company under the name DAN ^®, are suitable anionic surfactants.

Also, the sulfuric acid monoesters of straight-chain or branched C ₇ ethoxylated with 1 to 6 moles of ethylene oxide. ₂₁ -alcohols, such as 2-methyl-branched

with an average of 3.5 moles of ethylene oxide (EO) or C ₁₂ . ₁₈ fatty alcohols with 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 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 which are monoesters and / or diesters of sulfosuccinic acid with alcohols, preferably fatty alcohols and in particular ethoxylated fatty alcohols. Preferred sulfosuccinates contain C ₈ --i ₈ -fatty alcohol residues or mixtures of these. Particularly preferred sulfosuccinates contain a fatty alcohol residue derived from ethoxylated fatty alcohols, which by themselves are nonionic surfactants. 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 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.

Instead of the surfactants mentioned or in conjunction with them, it is also possible to use cationic and / or amphoteric surfactants.

As cationic active substances, for example, cationic compounds of the following formulas can be used:

Ri

Ri-N- (CH ₂₎ _n -T-R 2 (CH ₂₎ _n -T-R2

wherein each group R ^{1 is} independently selected from C-. ₆ alkyl, alkenyl or hydroxyalkyl groups; each R ² group is independently selected from C _{_8-28} alkyl or alkenyl groups; R ³ = R ¹ or (CH ₂ ) _n -TR ² ; R ⁴ = R ¹ or R ² or (CH ₂ ) _n -TR ² ; T = -CH ₂ -, -O-CO- or -CO-O- and n is an integer from 0 to 5.

In automatic dishwashing detergents, the content of cationic and / or amphoteric surfactants is preferably less than 6% by weight, preferably less than 4% by weight, very particularly preferably less than 2% by weight and in particular less than 1% by weight. %. Automatic dishwashing detergents containing no cationic or amphoteric surfactants are particularly preferred.

polymers The group of polymers includes, in particular, the washing or cleaning-active polymers, for example the rinse aid polymers and / or polymers which act as softeners. In general, cationic, anionic and amphoteric polymers can be used in detergents or cleaners in addition to nonionic polymers.

"Cationic polymers" for the purposes of the present invention are polymers which carry a positive charge in the polymer molecule, which can be realized, for example, by (alkyl) ammonium groups or other positively charged groups present in the polymer chain quaternized cellulose derivatives, the polysiloxanes with quaternary groups, the cationic guar derivatives, the polymeric dimethyldiallylammonium salts and their copolymers with esters and amides of acrylic acid and methacrylic acid, the copolymers of vinylpyrrolidone with quaternized derivatives of dialkylaminoacrylate and methacrylate, the vinylpyrrolidone-methoimidazolinium chloride Copolymers, the quaternized polyvinyl alcohols or the polymers listed under the INCI names Polyquaternium 2, Polyquaternium 17, Polyquaternium 18 and Polyquaternium 27.

For the purposes of the present invention, "amphoteric polymers" further comprise, in addition to a positively charged group in the polymer chain, also negatively charged groups or monomer units. These groups may be, for example, carboxylic acids, sulfonic acids or phosphonic acids.

Preferred washing or cleaning agents, in particular preferred automatic dishwashing agents, are characterized in that they contain a polymer a) which has monomer units of the formula R ¹ R ² C =CR ³ R ⁴ in which each R ¹ , R ² , R ⁴ radical ³ , R ^{4 is} independently selected from hydrogen, derivatized hydroxy, Ci- ₃₀ linear or branched alkyl groups, aryl, aryl substituted Ci _-30 linear or branched alkyl groups, polyalkoyxylierte alkyl groups, heteroatomic organic groups having at least one positive charge without charged nitrogen, at least a quaternized N atom or at least one amino group having a positive charge in the partial range of the pH range of 2 to 11, or salts thereof, with the proviso that at least one R ¹ , R ² , R ³ , R ^{4 is} a heteroatomic organic Group having at least one positive charge without charged nitrogen, at least one quaternized N atom or at least one aminog is a group with a positive charge.

In the context of the present application, particularly preferred cationic or amphoteric polymers contain as monomer unit a compound of the general formula

in which R ¹ and R ⁴ are each independently H or a linear or branched hydrocarbon radical having 1 to 6 carbon atoms; R ² and R ^{3 are} independently an alkyl, hydroxyalkyl, or aminoalkyl group in which the alkyl group is linear or branched and has from 1 to 6 carbon atoms, preferably a methyl group; x and y independently represent integers between 1 and 3. X represents a counterion, preferably a counterion selected from the group consisting of chloride, bromide, iodide, sulfate, hydrogensulfate, methosulfate, laurylsulfate, dodecylbenzenesulfonate, p-toluenesulfonate (tosylate), cumene sulfonate, xylenesulfonate, phosphate, citrate, formate, acetate or mixtures thereof.

Preferred radicals R ¹ and R ⁴ in the above formula are selected from -CH ₃ , -CH ₂ -CH ₃ , -CH ₂ -CH ₂ -CH ₃ , -CH (CH ₃ ) -CH ₃ , -CH ₂ -OH , -CH ₂ -CH ₂ -OH, -CH (OH) -CH _3, -CH ₂ -CH ₂ -CH ₂ -OH, -CH ₂ - CH (OH) -CH _3, -CH (OH) -CH ₂ -CH ₃ , and - (CH ₂ CH ₂ -O) _n H.

Very particular preference is given to polymers which have a cationic monomer unit of the above general formula in which R ¹ and R ^{4 are} H, R ² and R ^{3 are} methyl and x and y are each 1. The corresponding monomer unit of the formula

H ₂ C = C H- (CH ₂ ) -N ⁺ (CH ₃ ) ₂ - (CH ₂ ) -CH = CH ₂ X ^"

in the case of X = chloride, they are also referred to as DADMAC (diallyldimethylammonium chloride).

Further particularly preferred cationic or amphoteric polymers contain a monomer unit of the general formula

R1 HC = CR2-C (O) -NH- (CH ₂₎ -N R3R4R5

X ^" in the R ¹ , R ² , R ³ , R ⁴ and R ^{5 are} independently of one another a linear or branched, saturated or unsaturated alkyl or hydroxyalkyl radical having 1 to 6 carbon atoms, preferably a linear or branched alkyl radical selected from CH ₃ , -CH ₂ -CH ₃ , -CH ₂ - CH ₂ -CH ₃ , -CH (CH ₃ ) -CH ₃ , -CH ₂ -OH, -CH ₂ -CH ₂ -OH, -CH (OH) -CH ₃ , -CH ₂ -CH ₂ -CH ₂ -OH, -CH ₂ - CH (OH) -CH ₃ , -CH (OH) -CH ₂ -CH ₃ , and - (CH ₂ CH ₂ -O) _n H and x is an integer between 1 and 6.

For the purposes of the present application, very particular preference is given to polymers which have a cationic monomer unit of the above general formula in which R ^{1 is} H and R ² , R ³ , R ⁴ and R ^{5 are} methyl and x is 3. The corresponding monomer units of the formula

H ₂ C = C (CH ₃ ) -C (O) -NH- (CH 2) _X -N ⁺ (CH 3) 3

X ^"

in the case of X = chloride also referred to as MAPTAC (Methyacrylamidopropyl trimethylammonium chloride).

According to the invention, preference is given to using polymers which contain diallyldimethylammonium salts and / or acrylamidopropyltrimethylammonium salts as monomer units.

The aforementioned amphoteric polymers have not only cationic groups but also anionic groups or monomer units. Such anionic monomer units are derived, for example, from the group of linear or branched, saturated or unsaturated carboxylates, linear or branched, saturated or unsaturated phosphonates, linear or branched, saturated or unsaturated sulfates or linear or branched, saturated or unsaturated sulfonates. Preferred monomer units are acrylic acid, (meth) acrylic acid, (dimethyl) acrylic acid, (ethyl) acrylic acid, cyanoacrylic acid, vinylessingic acid, allylacetic acid, crotonic acid, maleic acid, fumaric acid, cinnamic acid and its derivatives, allylsulfonic acids such as allyloxybenzenesulfonic acid and methallylsulfonic acid or the allylphosphonic acids.

Preferred usable amphoteric polymers are selected from the group of the alkylacrylamide / acrylic acid copolymers, the alkylacrylamide / methacrylic acid copolymers, the alkylacrylamide / methylmethacrylic acid copolymers, the alkylacrylamide / acrylic acid / alkylaminoalkyl (meth) acrylic acid copolymers, the

Alkylacrylamide / methacrylic acid / alkylaminoalkyl (meth) acrylic acid copolymers, the alkylacrylamide / methylmethacrylic acid / alkylaminoalkyl (meth) acrylic acid copolymers, the alkylacrylamide / alkymethacrylate / alkylaminoethyl methacrylate / alkyl methacrylate copolymers and the copolymers of unsaturated carboxylic acids, cationically derivatized unsaturated carboxylic acids and optionally further ionic or nonionic monomers. Preferred zwitterionic polymers are from the group of acrylamidoalkyltrialkylammonium chloride / acrylic acid copolymers and their alkali metal and ammonium salts, the acrylamidoalkyltrialkylammonium chloride / methacrylic acid copolymers and their alkali metal and ammonium salts and the methacroylethylbetaine / methacrylate copolymers.

Also preferred are amphoteric polymers which comprise, in addition to one or more anionic monomers as cationic monomers, methacrylamidoalkyltrialkylammonium chloride and dimethyl (diallyl) ammonium chloride.

Particularly preferred amphoteric polymers are selected from the group of the methacrylamidoalkyltrialkylammonium chloride / dimethyl (diallyl) ammonium chloride / acrylic acid copolymers, the methacrylamidoalkyltrialkylammonium chloride / dimethyl (diallyl) ammonium chloride / methacrylic acid copolymers and the methacrylamidoalkyltrialkylammonium chloride / dimethyl (diallyl) ammonium chloride / alkyl ( meth) acrylic acid copolymers and their alkali metal and ammonium salts. Particular preference is given to amphoteric polymers from the group of the methacrylamidopropyltrimethylammonium chloride / dimethyl (diallyl) ammonium chloride / acrylic acid copolymers, the methacrylamidopropyltrimethylammonium chloride / dimethyldiallyl ammonium chloride / acrylic acid copolymers and the methacrylamidopropyltrimethylammonium chloride / dimethyl (diallyl) ammonium chloride / alkyl (meth ) acrylic acid copolymers and their alkali metal and ammonium salts.

In a particularly preferred embodiment of the present invention, the polymers are present in prefabricated form. For the preparation of the polymers, inter alia, the encapsulation of the polymers by means of water-soluble or water-dispersible coating compositions, preferably by means of water-soluble or water-dispersible natural or synthetic polymers; the encapsulation of the polymers by means of water-insoluble, meltable coating compositions, preferably by means of water-insoluble coating agents from the group of waxes or paraffins having a melting point above 30 ⁰ C; the co-granulation of the polymers with inert carrier materials, preferably with carrier materials from the group of washing- or cleaning-active substances, more preferably from the group of builders or cobuilders.

Detergents or cleaning agents contain the aforementioned cationic and / or amphoteric polymers preferably in amounts of between 0.01 and 10 wt .-%, each based on the total weight of the detergent or cleaning agent. In the context of the present application, however, preference is given to those detergents or cleaners in which the weight fraction of the cationic and / or amphoteric polymers is between 0.01 and 8% by weight, preferably between 0.01 and 6% by weight, preferably between 0.01 and 4 wt .-%, especially preferably between 0.01 and 2 wt .-% and in particular between 0.01 and 1 wt .-%, each based on the total weight of the automatic dishwashing detergent is.

Effective polymers as softeners are, for example, the sulfonic acid-containing polymers which are used with particular preference.

Particularly preferred as sulfonic acid-containing polymers are copolymers of unsaturated carboxylic acids, sulfonic acid-containing monomers and optionally other ionic or nonionic monomers.

In the context of the present invention are as unsaturated monomer carboxylic acids of the formula

R ¹ (R ² ) C = C (R ³ ) COOH

in which R ¹ to R ³ independently of one another are -H, -CH ₃ , a straight-chain or branched saturated alkyl radical having 2 to 12 carbon atoms, a straight-chain or branched, mono- or polyunsaturated alkenyl radical having 2 to 12 carbon atoms, NH ₂ , -OH or -COOH substituted alkyl or alkenyl radicals or -COOH or -COOR ⁴ , wherein R ^{4 is} a saturated or unsaturated, straight-chain or branched hydrocarbon radical having 1 to 12 carbon atoms.

Among the unsaturated carboxylic acids which can be described by the above formula are in particular acrylic acid (R = R = R = H), methacrylic acid (R = R = H, R = CH ₃ ) and / or maleic acid (R ¹ = COOH; R ² = R ³ = H) is preferred.

In the sulfonic acid-containing monomers are those of the formula

R ⁵ (R ⁶ ) C = C (R ⁷ ) -X-SO ₃ H

in which R ⁵ to R ⁷ independently of one another are -H, -CH ₃ , a straight-chain or branched saturated alkyl radical having 2 to 12 carbon atoms, a straight-chain or branched, mono- or polyunsaturated alkenyl radical having 2 to 12 carbon atoms, NH _2, -OH or -COOH-substituted alkyl or alkenyl, or -COOH or -COOR ⁴ wherein R ⁴ is a saturated or unsaturated, linear or branched hydrocarbon radical having 1 to 12 carbon atoms, and X is an optionally present spacer , which is selected from - (CH ₂ ), - - where n = 0 to 4, -COO- (CH ₂ ) _k - with k = 1 to 6, -C (O) -NH-C (CH ₃ ) ₂ - and -C (O) -NH-CH (CH ₂ CH ₃ ) -.

Preferred among these monomers are those of the formulas H ₂ C = CH-X-SO ₃ H

H ₂ C = C (CH 3) -X-SO ₃ H

HO ₃ SX (R ⁶⁾ C = C (R ⁷⁾ -X-SO ₃ H

in which R ⁶ and R ^{7 are} independently selected from -H, -CH ₃ , -CH ₂ CH ₃ , -CH ₂ CH ₂ CH ₃ , -CH (CH ₃ ) ₂ and X is an optional spacer group which is selected from - (CHz) _n - with n = 0 to 4, -COO- (CH ₂ ) _k - with k = 1 to 6, -C (O) -NH-C (CH ₃ ) ₂ - and -C (O) -NH-CH (CH ₂ CH ₃ ) -.

Particularly preferred monomers containing sulfonic acid groups are 1-acrylamido-1-propanesulfonic acid, 2-acrylamido-2-propanesulfonic acid, 2-acrylamido-2-methyl-1-propanesulfonic acid, 2-methacrylamido-2-methyl 1-propanesulfonic acid, 3-methacrylamido-2-hydroxypropanesulfonic acid, allylsulfonic acid, methallylsulfonic acid, allyloxybenzenesulfonic acid, methallyloxybenzenesulfonic acid, 2-hydroxy-3- (2-propenyloxy) propanesulfonic acid, 2-methyl-2-propenylsulfonic acid, styrenesulfonic acid, vinylsulfonic acid, 3-sulfopropyl acrylate, 3-sulfopropyl methacrylate, sulfomethacrylamide, sulfomethylmethacrylamide and water-soluble salts of said acids.

Particularly suitable other ionic or nonionic monomers are ethylenically unsaturated compounds. The content of the polymers used in these other ionic or nonionic monomers is preferably less than 20% by weight, based on the polymer. Particularly preferred polymers to be used consist only of monomers of the formula R ¹ (R ² ) C =C (R ³ ) COOH and monomers of the formula R ⁵ (R ⁶ ) C =C (R ⁷ ) -

X-SO ₃ H.

Further particularly preferred copolymers consist of i) one or more unsaturated carboxylic acids from the group of acrylic acid,

Methacrylic acid and / or maleic acid ii) one or more sulfonic acid group-containing monomers of the formulas:

H ₂ C = CH-X-SO ₃ H

H ₂ C = C (CH ₃ ) -X-SO ₃ H

HO ₃ SX (R ⁶⁾ C = C (R ⁷⁾ -X-SO ₃ H

in which R ⁶ and R ^{7 are} independently selected from -H, -CH ₃ , -CH ₂ CH ₃ , -CH ₂ CH ₂ CH ₃ , -CH (CH ₃ ) ₂ and X is an optional spacer group which is selected from - (CHz) _n - with n = 0 to 4, -COO- (CH ₂ ) _k - with k = 1 to 6, -C (O) -NH-C (CHs) ₂ - and -C ( O) -NH-CH (CH ₂ CH ₃ ) - iii) optionally further ionogenic or nonionic monomers.

The copolymers may contain the monomers from groups i) and ii) and, if appropriate, iii) in varying amounts, it being possible for all representatives from group i) to be combined with all representatives from group ii) and all representatives from group iii). Particularly preferred polymers have certain structural units, which are described below.

For example, copolymers which are structural units of the formula are preferred

- [CH ₂ -CHCOOH] _m - [CH ₂ -CHC (O) -Y-SO ₃ H] _p -

in which m and p are each an integer between 1 and 2,000 and Y is a spacer group selected from substituted or unsubstituted aliphatic, aromatic or substituted aromatic hydrocarbon radicals having 1 to 24 carbon atoms, wherein spacer groups in which Y. for -O- (CH ₂ ) _n - with n = 0 to 4, for -O- (C ₆ H ₄ ) -, for -NH-C (CHs) ₂ - or -NH-CH (CH ₂ CH ₃ ) - stands, are preferred.

These polymers are prepared by copolymerization of acrylic acid with a sulfonic acid-containing acrylic acid derivative. When the acrylic acid derivative containing sulfonic acid groups is copolymerized with methacrylic acid, another polymer is obtained whose use is likewise preferred. The corresponding copolymers contain the structural units of the formula

- [CH ₂ -C (CH ₃ ) COOH] _m - [CH ₂ -CHC (O) -Y-SO ₃ H] _p -

in which m and p are each an integer between 1 and 2000 and Y is a spacer group selected from substituted or unsubstituted aliphatic, aromatic or substituted aromatic hydrocarbon radicals having 1 to 24 carbon atoms, wherein O- (CH ₂ ) _n - where n = 0 to 4, -O- (C ₆ H ₄ ) -, -NH-C (CHs) ₂ - or -NH-CH (CH ₂ CH ₃ ) - , are preferred.

Acrylic acid and / or methacrylic acid can also be copolymerized completely analogously with methacrylic acid derivatives containing sulfonic acid groups, as a result of which the structural units in the molecule are changed. Thus, copolymers which are structural units of the formula

- [CH ₂ -CHCOOH] _m - [CH ₂ -C (CH ₃ ) C (O) -Y-SO ₃ H] _p - in which m and p are each an integer between 1 and 2,000 and Y is a spacer group selected from substituted or unsubstituted aliphatic, aromatic or substituted aromatic hydrocarbon radicals having 1 to 24 carbon atoms, wherein spacer groups in which Y. for -O- (CH ₂ ) _n - with n = 0 to 4, for -O- (C ₆ H ₄ ) -, for -NH-C (CH ₃ ) ₂ - or -NH-CH (CH ₂ CH ₃ ) -, are particularly preferred, just as preferred as copolymers, the structural units of the formula

- [CH ₂ -C (CH ₃ ) COOH] ^ [CH ₂ -C (CH ₃ ) C (O) -Y-SO ₃ H] _P -

Instead of acrylic acid and / or methacrylic acid or in addition thereto, maleic acid can also be used as a particularly preferred monomer from group i). This gives way to inventively preferred copolymers, the structural units of the formula

- [HOOCCH-CHCOOH] _m - [CH ₂ -CHC (O) -Y-SO ₃ H] _p -

in which m and p are in each case an integer from 1 to 2000 and Y is a spacer group selected from substituted or unsubstituted aliphatic, aromatic or araliphatic hydrocarbon radicals having from 1 to 24 carbon atoms, wherein spacer groups in which Y represents -O- (CH ₂ ) _n - with n = 0 to 4, for -O- (C ₆ H ₄ ) -, for -NH-C (CH ₃ ) ₂ - or -NH-CH (CH ₂ CH ₃ ) - stands, are preferred. Also preferred according to the invention are copolymers which contain structural units of the formula

- [HOOCCH-CHCOOH] _m - [CH ₂ -C (CH ₃ ) C (O) OY-SO ₃ H] _p -

in which m and p are each an integer between 1 and 2,000 and Y is a spacer group selected from substituted or unsubstituted aliphatic, aromatic or substituted aromatic hydrocarbon radicals having 1 to 24 carbon atoms, wherein spacer groups in which Y. for -O- (CH ₂ ) _n - with n = 0 to 4, for -O- (C ₆ H ₄ ) -, for -NH-C (CHs) ₂ - or -NH-CH (CH ₂ CH ₃ ) - stands.

In the polymers, the sulfonic acid groups may be wholly or partially in neutralized form, ie that the acidic acid of the sulfonic acid group in some or all sulfonic acid groups against metal ions, preferably alkali metal ions and in particular against Sodium ions, can be replaced. The use of partially or fully neutralized sulfonic acid-containing copolymers is preferred according to the invention.

The monomer distribution of the copolymers preferably used according to the invention in the case of copolymers which contain only monomers from groups i) and ii) is preferably in each case from 5 to 95% by weight i) or ii), particularly preferably from 50 to 90% by weight monomer from group i) and from 10 to 50% by weight of monomer from group ii), in each case based on the polymer.

In the case of terpolymers, particular preference is given to those containing from 20 to 85% by weight of monomer from group i), from 10 to 60% by weight of monomer from group ii) and from 5 to 30% by weight of monomer from group iii) ,

The molar mass of the sulfo copolymers preferably used according to the invention can be varied in order to adapt the properties of the polymers to the desired end use. Preferred washing or cleaning agents are characterized in that the copolymers have molar masses of 2000 to 200,000 gmol ^"1 , preferably from 4000 to 25,000 gmol ^'1 and in particular from 5000 to 15,000 gmol ^{" 1} .

bleach

The bleaching agents are a particularly preferred washing or cleaning substance. Among the compounds serving as bleaches in water H ₂ O ₂ , sodium percarbonate, sodium perborate tetrahydrate and sodium perborate monohydrate are of particular importance. Other useful bleaching agents are, for example, peroxypyrophosphates, citrate perhydrates and H ₂ O ₂ -forming peracidic salts or peracids, such as perbenzoates, peroxophthalates, diperazelaic acid, phthaloiminoperacid or diperdodecanedioic acid.

Furthermore, bleaching agents from the group of organic bleaching agents can also be used. 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 (a) the peroxybenzoic acid and its ring-substituted derivatives, such as alkylperoxybenzoic acids, but also peroxy-α-naphthoic acid and magnesium monoperphthalate, (b) the aliphatic or substituted aliphatic peroxyacids, such as peroxylauric acid, peroxystearic acid, ε-phthalimidoperoxycaproic acid [phthaliminoperoxyhexanoic acid (PAP)] , o- Carboxybenzamidoperoxycaproic acid, N-Nonenylamidoperadipinsäure and N-Nonenylamidopersuccinate, and (c) aliphatic and araliphatic Peroxyd icarbonsäuren, such as 1, 12-diperoxycarboxylic acid, 1, 9-Diperoxyazelainsäure, Diperocysebacinsäure, Diperoxybrassylic acid, the diperoxyphthalic acids, 2-decyldiperoxybutane-1,4-diacid, N, N-terephthaloyl-di (6-aminopercapronate).

As a bleaching agent and chlorine or bromine releasing substances can be used. Among the suitable chlorine or bromine releasing materials are for example heterocyclic N-bromo- and N-chloroamides, for example trichloroisocyanuric acid, tribromoisocyanuric acid, dibromoisocyanuric acid and / or dichloroisocyanuric acid (DICA) and / or their salts with cations such as potassium and sodium into consideration. Hydantoin compounds, such as 1,3-dichloro-5,5-dimethylhydantoin are also suitable.

According to the invention, washing or cleaning agents, in particular automatic dishwashing agents, are preferred which contain from 1 to 35% by weight, preferably from 2.5 to 30% by weight, particularly preferably from 3.5 to 20% by weight and in particular from 5 to 15% by weight % Bleach, preferably sodium percarbonate.

The active oxygen content of the washing or cleaning agents, in particular the automatic dishwashing agents, in each case based on the total weight of the composition, preferably between 0.4 and 10 wt .-%, particularly preferably between 0.5 and 8 wt .-% and in particular between 0.6 and 5 wt .-%. Particularly preferred compositions have an active oxygen content above 0.3 wt .-%, preferably above 0.7 wt .-%, more preferably above 0.8 wt .-% and in particular above 1, 0 wt .-% to.

Bleach activators

Bleach activators are used in detergents or cleaners, for example, to achieve an improved bleaching effect when cleaning at temperatures of 60 ⁰ C and below. 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 tetraacetylglycoluril (TAGU), N- Acylimides, in particular N-nonanoylsuccinimide (NOSI), acylated phenolsulfonates, in particular n-nonanoyl or isononanoyloxybenzenesulfonate (n- or iso-NOBS), carboxylic anhydrides, in particular phthalic anhydride, acylated polyhydric alcohols, in particular triacetin, ethylene glycol diacetate and 2,5-diacetoxy- 2,5-dihydrofuran, n-methyl-morpholinium-acetonitrile-methyl sulfate (MMA) and acetylated sorbitol and mannitol or mixtures thereof (SORMAN), acylated sugar derivatives, in particular Pentaacetyl glucose (PAG), pentaacetyl fructose, tetraacetylxylose and octaacetyl lactose, and acetylated, optionally N-alkylated glucamine and gluconolactone, and / or N-acylated lactams, for example N-benzoyl caprolactam. Hydrophilic substituted acyl acetals and acyl lactams are also preferably used. Combinations of conventional bleach activators can also be used.

These bleach activators are preferably used in amounts of up to 10% by weight, in particular 0.1% by weight to 8% by weight, especially 2 to 8% by weight and more preferably 2 to 6% by weight, based in each case on the total weight of bleach activator-containing agents.

Further bleach activators preferably used in the context of the present application are compounds from the group of cationic nitriles, in particular cationic nitriles of the formula

Ri

R2-N- (CH ₂₎ -cnx ^"

R3

in which R ¹ is -H, -CH _3, a C ₂ - ₂₄ alkyl or alkenyl group, a substituted C _{_2-24} -alkyl or -alkenyl radical having at least one substituent from the group -Cl, -Br, - OH, -NH _2, -CN, an alkyl or alkenylaryl radical with a CI_ ₂₄ alkyl group, or a substituted alkyl- or alkenylaryl radical with a CI_ ₂₄ alkyl group and at least one further substituent on the aromatic ring, R ² and R ³ are independently selected from -CH ₂ -CN, -CH ₃ , -CH ₂ -CH ₃ , -CH ₂ -CH ₂ -CH ₃ , -CH (CH ₃ ) -CH ₃ , -CH ₂ -OH, -CH ₂ -CH ₂ -OH, -CH (OH) -CH ₃ , -CH ₂ -CH ₂ -CH ₂ -OH, -CH ₂ -CH (OH) -CH ₃ , -CH (OH) -CH ₂ -CH ₃ , - (CH ₂ CH ₂ -O) _n H where n = 1, 2, 3, 4, 5 or 6 and X is an anion.

Particularly preferred is a cationic nitrile of the formula

in which R ⁴ , R ⁵ and R ^{6 are} independently selected from -CH ₃ , -CH ₂ -CH ₃ , -CH ₂ -CH ₂ -CH ₃ , -CH (CH ₃ ) -CH ₃ , wherein R ⁴ additionally also -H may be and X is an anion, preferably R ⁵ = R ⁶ = -CH ₃ and in particular R ⁴ = R ⁵ = R ⁶ = -CH ₃ and compounds of the formulas (CH ₃ ) ₃ N ⁽⁺⁾ CH ₂ -CN X ^" , (CH ₃ CH ₂ ) ₃ N ⁽⁺⁾ CH ₂ -CN X ^" , (CH ₃ CH ₂ CH ₂ ) ₃ N ⁽⁺⁾ CH ₂ -CN X ^" , (CH ₃ CH ( CH ₃ )) ₃ N ⁽⁺⁾ CH ₂ -CN X ^" , or (HO-CH ₂ -CH ₂ ) ₃ N ⁽⁺⁾ CH ₂ -CN X ^{" are} particularly preferred, wherein from the group of these substances turn the cationic Nitrile of the formula (CH ₃ ) ₃ N ⁽⁺⁾ CH ₂ -CN X ^' in which X ^{' is} an anion selected from the group consisting of chloride, bromide, iodide, hydrogensulfate, methosulfate, p-toluenesulfonate (tosylate) or xylenesulfonate is particularly preferred.

bleach catalysts

In addition to the conventional bleach activators or in their place, so-called bleach catalysts can also be used. These substances are bleach-enhancing transition metal salts or transition metal complexes such as, for example, Mn, Fe, Co, Ru or Mo saline 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 can also be used as bleach catalysts.

Bleach-enhancing transition metal complexes, in particular having the central atoms Mn, Fe, Co, Cu, Mo, V, Ti and / or Ru, preferably selected from the group of the manganese and / or cobalt salts and / or complexes, particularly preferably the cobalt (ammine) Complexes, the cobalt (acetate) complexes, the cobalt (carbonyl) complexes, the chlorides of cobalt or manganese, of the manganese sulfate are in conventional amounts, preferably in an amount up to 5 wt .-%, in particular of 0.0025 Wt .-% to 1 wt .-% and particularly preferably from 0.01 wt .-% to 0.25 wt .-%, each based on the total weight of the inventive agents used. In special cases, however, more bleach activator can also be used.

It is particularly preferred to use complexes of manganese in the oxidation state II, IM, IV or IV, which preferably contain one or more macrocyclic ligands with the donor functions N, NR, PR, O and / or S. Preferably, ligands are used which have nitrogen donor functions. It is particularly preferred to use bleach catalyst (s) in the compositions of the invention, which as macromolecular ligands 1, 4,7-trimethyl-1, 4,7-triazacyclononan (Me-TACN), 1, 4,7-triazacyclononane (TACN ), 1, 5,9-trimethyl-1, 5,9-triazacyclododecane (Me-TACD), 2-methyl-1, 4,7-trimethyl-1, 4,7-triazacyclononane (Me / Me-TACN) and or 2-methyl-1, 4,7-triazacyclononane (Me / TACN). Suitable manganese complexes are, for example, [Mn ^m ₂ (μ-O) i (μ-OAc) ₂ (TACN) ₂ ] (CIO ₄ ) ₂ , [Mn ^m Mn ' ^v (μ-O) ₂ (μ-OAc) ₁ ( TACN) ₂ ] (BPh ₄ ) Z, [Mn ^IV 4 (μ-O) ₆ (TACN) 4] (Clθ4) 4, [Mn ^I " ₂ (μ-O) ₁ (μ-OAc) ₂ (MeOH) TACN) ₂ ] (CIO ₄ ) 2, [Mn " ¹ Mn ^lv (μ-O) ₁ (μ-OAc) ₂ (Me-TACN) ₂ ] (CIO ₄ ) 3, [Mn ' ^v ₂ (μ-O ) ₃ (Me-TACN) _2] (PF ₆₎ ₂ and [Mn ^'v ₂ (μ- O) ₍₃ (Me / Me-TACN) _2] PF ₆₎ ₂ (OAc = OC (O) CH ₃₎ ,

To increase the washing or cleaning performance of detergents or cleaners, the detergents and cleaners according to the invention may contain further enzymes contain. These include in particular proteases, amylases, cellulases, hemicellulases, oxidoreductases, preferably oxidases and perhydrolases and lipases, and preferably mixtures thereof. These enzymes are basically of natural origin; Starting from the natural molecules, improved variants are available for use in detergents and cleaners, which are preferably used accordingly. Detergents or cleaning agents contain enzymes preferably in total amounts of 1 × 10 ^-6 to 5 wt .-% based on active protein. The protein concentration can be determined by known methods, for example the BCA method or the biuret method. Methods for determining the protein concentration are well known to the person skilled in the art.

Among the proteases, those of the subtilisin type are preferable. Examples thereof are the subtilisins BPN 'and Carlsberg, the protease PB92, the subtilisins 147 and 309, the alkaline protease from Bacillus lentus, subtilisin DY and the enzymes thermitase, proteinase K and the subtilases, but not the subtilisins in the narrower sense Proteases TW3 and TW7. Subtilisin Carlsberg is available in a further developed form under the trade name Alcalase® from Novozymes A / S, Bagsvaerd, Denmark. The subtilisins 147 and 309 are sold under the trade names Esperase®, and Savinase® by the company Novozymes. From the protease from Bacillus lentus DSM 5483 derived under the name BLAP® protease variants derived.

Further useful proteases are, for example, those under the trade names Durazym®, Relase®, Everlase®, Nafizym, Natalase®, Kannase® and Ovozymes® from Novozymes, sold under the trade name, Purafect®, Purafect® OxP and Properase® by the company Genencor, sold under the trade name Protosol® by the company Advanced Biochemicals Ltd., Thane, India, under the trade name Wuxi® by Wuxi Snyder Bioproducts Ltd., China, under the trade names Proleather® and Protease P® by the company Amano Pharmaceuticals Ltd., Nagoya, Japan, and the enzyme available under the name Proteinase K-16 from Kao Corp., Tokyo, Japan.

Examples of amylases are the α-amylases from Bacillus licheniformis, from B. amyloliquefaciens or from B. stearothermophilus and their further developments for use in detergents or cleaners. The B. licheniformis enzyme is available from Novozymes under the name Termamyl® and from Genencor under the name Purastar®ST. Further development products of this α-amylase are available from Novozymes under the trade names Duramyl® and Termamyl®ultra, from Genencor under the name Purastar®OxAm, and from Daiwa Seiko Inc., Tokyo, Japan, as Keistase®. B. amyloliquefaciens α-amylase is sold by Novozymes under the name BAN® and derived variants of B. stearothermophilus α-amylase under the names BSG® and Novamyl®, likewise from Novozymes.

Furthermore, the α-amylase from Bacillus sp. A 7-7 (DSM 12368) and the cyclodextrin glucanotransferase (CGTase) from B. agaradherens (DSM 9948). Further, the amylolytic enzymes disclosed in International Patent Applications WO 03/002711 A2 and WO 03/054177 A2 can be used. Likewise, fusion products of all the molecules mentioned can be used.

In addition, the further developments of the α-amylase from Aspergillus niger and A. oryzae available under the trade names Fungamyl® from Novozymes are suitable. Further usable commercial products are, for example, the Amylase-LT® and Stainzyme® or Stainzyme ultra® or Stainzyme plus®, the latter also from Novozymes. Variants of these enzymes which can be obtained by point mutations can also be incorporated into agents according to the invention.

Agents of the invention may further contain lipases or cutinases, in particular because of their triglyceride-cleaving activities, but also to generate in situ peracids from suitable precursors. Examples of such enzymes are the lipases originally obtainable or further developed from Humicola lanuginosa (Thermomyces lanuginosus), in particular those with the amino acid exchange D96L. They are sold for example by the company Novozymes under the trade names Lipolase®, Lipolase®Ultra, LipoPrime®, Lipozyme® and Lipex®. Furthermore, for example, the cutinases can be used, which were originally isolated from Fusarium solani pisi and Humicola insolens. Lipases which are likewise useful are sold by Amano under the names Lipase CE®, Lipase P®, Lipase B® or Lipase CES®, Lipase AKG®, Bacillis sp. Lipase®, Lipase AP®, Lipase M-AP® and Lipase AML®. By Genencor, for example, the lipases or cutinases can be used, the initial enzymes were originally isolated from Pseudomonas mendocina and Fusarium solanii. Other important commercial products are the preparations M1 Lipase.RTM. And Lipomax.RTM. Originally sold by Gist-Brocades and the enzymes marketed by Meito Sangyo KK, Japan, under the name Lipase MY-30®, Lipase OF® and Lipase PL® to mention also the product Lumafast® from the company Genencor.

Cellulases, often also referred to as endoglucanases, can be used according to the invention, in particular if they are intended for the treatment of textiles, depending on the purpose as pure enzymes, as enzyme preparations or in the form of mixtures in which the individual components advantageously with regard to their different performance aspects complete. To These performance aspects include, in particular, contributions to the primary washing performance, the secondary washing performance of the composition (anti-redeposition effect or graying inhibition) and softening (fabric effect), up to the exercise of a "stone washed" effect.

A useful fungal, Endoglucanase (EC) -rich cellulase preparation, or their further developments are offered by the company Novozymes under the trade name Celluzyme®. The products Endolase® and Carezyme®, which are also available from Novozymes, are based on the 50 kD EG or the 43 kD EG from H. insolens DSM 1800. Other commercially available products of this company are Cellusoft® and Renozyme®. Further, for example, the 20 kD-EG from Melanocarpus, which is available from AB Enzymes, Finland, under the trade names Ecostone® and Biotouch®. Further commercial products of AB Enzymes are Econase® and Ecopulp®. Other suitable cellulases are from Bacillus sp. CBS 670.93 and CBS 669.93, those derived from Bacillus sp. CBS 670.93 from the company Genencor under the trade name Puradax® is available. Further commercial products of Genencor are "Genencor detergent cellulase L" and lndiAge®Neutra.

Examples of further enzymes, which are summarized by the term hemicellulases, are mannanases, xanthan lyases, pectin lyases (= pectinases), pectin esterases, pectate lyases, xyloglucanases (= xylanases), pullulanases and β-glucanases. Suitable enzymes for this purpose are, for example, under the name Gamanase® and Pektinex AR® from Novozymes, under the name Rohapec® B1 L from AB Enzymes and under the name Pyrolase® from Diversa Corp., San Diego, CA, USA available. The β-glucanase obtained from Bacillus subtilis is available under the name Cereflo® from Novozymes. Hemicellulases which are particularly preferred according to the invention are mannanases which are sold, for example, under the trade names Mannaway (R) by the company Novozymes or Purabrite (R) by the company Genencor.

Particular preference is given to using perhydrolases in the compositions according to the invention or enzymes which have a perhydrolase activity.

To increase the bleaching effect, further oxidoreductases, for example further oxidases, oxygenases, catalases, peroxidases, such as halo, chloro, bromo, lignin, glucose or manganese peroxidases, dioxygenases or laccases (phenol oxidases, polyphenol oxidases) can be used according to the invention , Advantageously, it is additionally preferred to add organic, particularly preferably aromatic, compounds which interact with the enzymes in order to enhance the activity of the relevant oxidoreductases (enhancers) or to ensure the flow of electrons (mediators) at greatly varying redox potentials between the oxidizing enzymes and the soils. The enzymes can be used in any form known in the art. These include, for example, the solid preparations obtained by granulation, extrusion or lyophilization or, especially in the case of liquid or gel-form detergents, solutions of the enzymes, advantageously as concentrated as possible, sparing in water and / or added with stabilizers.

Alternatively, the enzymes may be encapsulated for both the solid and liquid dosage forms, for example by spray-drying or extruding the enzyme solution together with a preferably natural polymer or in the form of capsules, for example those in which the enzymes are entrapped as in a solidified gel or in those of the core-shell type, in which an enzyme-containing core is coated with a water, air and / or chemical impermeable protective layer. In deposited layers, further active ingredients, for example stabilizers, emulsifiers, pigments, bleaches or dyes, may additionally be applied. Such capsules are applied by methods known per se, for example by shaking or rolling granulation or in fluid-bed processes. Advantageously, such granules, for example by applying polymeric film-forming agent, low in dust and storage stable due to the coating.

Furthermore, it is possible to assemble two or more enzymes together so that a single granule has several enzyme activities.

A protein and / or enzyme may be particularly protected during storage against damage such as inactivation, denaturation or degradation, such as by physical influences, oxidation or proteolytic cleavage. In microbial recovery of proteins and / or enzymes, inhibition of proteolysis is particularly preferred, especially if the agents also contain proteases. Detergents may contain stabilizers for this purpose; the provision of such means constitutes a preferred embodiment of the present invention.

Glass corrosion inhibitors

Glass corrosion inhibitors prevent the occurrence of haze, streaks and scratches, but also iridescence of the glass surface of machine-cleaned glasses. Preferred glass corrosion inhibitors come from the group of magnesium and zinc salts and magnesium and zinc complexes.

The spectrum of the preferred zinc salts according to the invention, preferably organic acids, particularly preferably organic carboxylic acids, ranges from salts which are difficult or insoluble in water, ie a solubility below 100 mg / l, preferably below 10 mg / l, in particular below 0.01 mg / l, to those salts which have a solubility in water above 100 mg / l, preferably above 500 mg / l, more preferably above 1 g / l and especially above 5 g / l (all Solubilities at 2O ⁰ C water temperature). The first group of zinc salts includes, for example, the zinc nitrate, the zinc oleate and the zinc stearate, and the group of soluble zinc salts includes, for example, zinc formate, zinc acetate, zinc lactate and zinc gluconate.

With particular preference is used as the glass corrosion inhibitor at least one zinc salt of an organic carboxylic acid, more preferably a zinc salt from the group zinc stearate, zinc oleate, zinc gluconate, zinc acetate, zinc lactate and Zinkeitrat used. Zinc ricinoleate, zinc abietate and zinc oxalate are also preferred.

In the context of the present invention, the content of zinc salt in detergents or cleaning agents is preferably between 0.1 and 5 wt.%, Preferably between 0.2 and 4 wt.% And in particular between 0.4 and 3 wt. , or the content of zinc in oxidized form (calculated as Zn ²⁺ ) between 0.01 to 1 wt .-%, preferably between 0.02 to 0.5 wt .-% and in particular between 0.04 to 0, 2 wt .-%, each based on the total weight of the glass corrosion inhibitor-containing agent.

Corrosion inhibitors serve to protect the items to be washed or the machine, with particular silver protectants being of particular importance in the field of automatic dishwashing. It is possible to use the known substances of the prior art. In general, silver protectants selected from the group of triazoles, benzotriazoles, bisbenzotriazoles, aminotriazoles, alkylaminotriazoles and transition metal salts or complexes can be used in particular. Particularly preferred to use are benzotriazole and / or alkylaminotriazole. According to the invention, preference is given to using 3-amino-5-alkyl-1,2,4-triazoles or their physiologically tolerated salts, these substances being particularly preferably present in a concentration of 0.001 to 10% by weight, preferably 0.0025 to 2 Wt .-%, particularly preferably 0.01 to 0.04 wt .-% are used. Preferred acids for salt formation are hydrochloric acid, sulfuric acid, phosphoric acid, carbonic acid, sulphurous acid, organic carboxylic acids such as acetic, glycolic, citric, succinic acid. Especially effective are 5-pentyl, 5-heptyl, 5-nonyl, 5-undecyl, 5-isononyl, 5-versatic-10-alkyl-3-amino-1, 2,4-triazoles and mixtures of these substances.

In addition, cleaner formulations often contain active chlorine-containing agents which can markedly reduce the corrosion of the silver surface. In chlorine-free cleaners especially oxygen and nitrogen-containing organic redox-active compounds, such as di- and trihydric phenols, such as hydroquinone, pyrocatechol, hydroxyhydroquinone, gallic acid, phloroglucinol, pyrogallol or derivatives of these classes of compounds are used. Also salt and Complex inorganic compounds such as salts of the metals Mn, Ti, Zr, Hf, V, Co and Ce are frequently used. Preferred here are the transition metal salts, which are selected from the group of manganese and / or cobalt salts and / or complexes, particularly preferably the cobalt (amnine) -Konnplexe, the cobalt (acetate) complexes, the cobalt (carbonyl) - Complexes, the chlorides of cobalt or manganese and manganese sulfate. Also, zinc compounds can be used to prevent corrosion on the items to be washed.

Instead of or in addition to the silver protectants described above, for example the benzotriazoles, redox-active substances can be used. These substances are preferably inorganic redox-active substances from the group of manganese, titanium, zirconium, hafnium, vanadium, cobalt and cerium salts and / or complexes, wherein the metals preferably in one of the oxidation states II, IM, IV, V or VI are present.

The metal salts or metal complexes used should be at least partially soluble in water. The counterions suitable for salt formation include all conventional mono-, di-, or tri-negatively charged inorganic anions, e.g. Oxide, sulfate, nitrate, fluoride, but also organic anions such as e.g. Stearate.

Particularly preferred metal salts and / or metal complexes are selected from the group MnSO ₄ , Mn (II) citrate, Mn (II) stearate, Mn (II) acetylacetonate, Mn (II) - [1-hydroxyethane-1, 1- diphosphonate], V ₂ O ₅ , V ₂ O ₄ , VO ₂ , TiOSO ₄ , K ₂ TiF ₆ , K ₂ ZrF ₆ , CoSO ₄ , Co (NO ₃ ) ₂ , Ce (NO ₃ ) ₃ , and mixtures thereof, such that the metal salts and / or metal complexes are selected from the group MnSO ₄ , Mn (II) citrate, Mn (II) stearate, Mn (II) acetylacetonate, Mn (II) - [1-hydroxyethane-1, 1- diphosphonate], V ₂ O ₅ , V ₂ O ₄ , VO ₂ , TiOSO ₄ , K ₂ TiF ₆ , K ₂ ZrF ₆ , CoSO ₄ , Co (NO ₃ ) ₂ , Ce (NO ₃ ) _{3 are used} with particular preference ,

The inorganic redox-active substances, in particular metal salts or metal complexes are preferably coated, i. completely coated with a waterproof, but easily soluble in the cleaning temperatures material to prevent their premature decomposition or oxidation during storage. Preferred coating materials, which are applied by known methods, such as Sandwik from the food industry, are paraffins, microwaxes, waxes of natural origin such as carnauba wax, candellila wax, beeswax, higher melting alcohols such as hexadecanol, soaps or fatty acids.

The metal salts and / or metal complexes mentioned are contained in cleaning agents, preferably in an amount of 0.05 to 6 wt .-%, preferably 0.2 to 2.5 wt .-%, each based on the total agent. In order to facilitate the disintegration of preformed moldings, it is possible to incorporate disintegration aids, so-called tablet disintegrants, into these compositions in order to shorten the disintegration times. By tablet disintegrants or disintegrants are meant auxiliaries which ensure the rapid disintegration of tablets in water or other media and for the release of the active ingredients.

These substances, which are also referred to as "explosives" due to their effect, increase their volume upon ingress of water, on the one hand increasing the intrinsic volume (swelling), and on the other hand creating a pressure via the release of gases which can break the tablet into smaller particles decays. Well-known disintegration aids are, for example, carbonate / citric acid systems, although other organic acids can also be used. Swelling disintegration aids are, for example, synthetic polymers such as polyvinylpyrrolidone (PVP) or natural polymers or modified natural substances such as cellulose and starch and their derivatives, alginates or casein derivatives.

Disintegration aids are preferably used in amounts of from 0.5 to 10% by weight, preferably from 3 to 7% by weight and in particular from 4 to 6% by weight, based in each case on the total weight of the disintegration assistant-containing agent.

Preferred disintegrating agents are cellulosic disintegrating agents, so that preferred washing or cleaning agents comprise such cellulose-based disintegrants in amounts of from 0.5 to 10% by weight, preferably from 3 to 7% by weight and in particular from 4 to 6% by weight. % contain. Pure cellulose has the formal gross composition (C ₆ H ₁₀ Os) _n and is formally a β-1,4-polyacetal of cellobiose, which in turn is composed of two molecules of glucose. Suitable celluloses consist of about 500 to 5000 glucose units and therefore have average molecular weights of 50,000 to 500,000. Cellulose-based disintegrating agents which can be used in the context of the present invention are also cellulose derivatives obtainable by polymer-analogous reactions of cellulose. Such chemically modified celluloses include, for example, products of esterifications or etherifications in which hydroxy hydrogen atoms have been substituted. Celluloses in which the hydroxy groups have been replaced by functional groups which are not bonded via an oxygen atom can also be used as cellulose derivatives. The group of cellulose derivatives includes, for example, alkali metal celluloses, carboxymethylcellulose (CMC), cellulose esters and ethers, and aminocelluloses. The mentioned cellulose derivatives are preferably not used alone as disintegrating agents based on cellulose, but used in admixture with cellulose. The content of these mixtures of cellulose derivatives is preferably below 50 wt .-%, more preferably below 20 wt .-%, based on the Cellulose-based disintegrant. It is particularly preferred to use cellulose-based disintegrating agent which is free of cellulose derivatives.

The cellulose used as a disintegration aid is preferably not used in finely divided form, but converted into a coarser form, for example granulated or compacted, before it is added to the premixes to be tabletted. The particle sizes of such disintegrating agents are usually above 200 .mu.m, preferably at least 90 wt .-% between 300 and 1600 .mu.m and in particular at least 90 wt .-% between 400 and 1200 microns.

As a further disintegrating agent based on cellulose or as a component of this component microcrystalline cellulose can be used. This microcrystalline cellulose is obtained by partial hydrolysis of celluloses under conditions which attack and completely dissolve only the amorphous regions (about 30% of the total cellulose mass) of the celluloses, leaving the crystalline regions (about 70%) intact. Subsequent deaggregation of the microfine celluloses produced by the hydrolysis yields the microcrystalline celluloses which have primary particle sizes of about 5 μm and can be compacted, for example, into granules having an average particle size of 200 μm.

Preferred disintegration aids, preferably a disintegration aid based on cellulose, preferably in granular, cogranulated or compacted form, are present in the disintegrating agent-containing agents in amounts of from 0.5 to 10% by weight, preferably from 3 to 7% by weight and in particular from 4 to 6 wt .-%, each based on the total weight of the disintegrating agent-containing agent.

Furthermore, according to the invention, gas-evolving effervescent systems can furthermore be used as tablet disintegration auxiliaries. The gas-evolving effervescent system may consist of a single substance that releases a gas upon contact with water. Among these compounds, mention should be made in particular of magnesium peroxide, which liberates oxygen on contact with water. Usually, however, the gas-releasing effervescent system in turn consists of at least two constituents which react with one another to form gas. While a variety of systems are conceivable and executable here, which release, for example, nitrogen, oxygen or hydrogen, the bubble system used in detergents and cleaners can be selected both on the basis of economic and environmental considerations. Preferred effervescent systems consist of alkali metal carbonate and / or bicarbonate and an acidifying agent which is suitable for liberating carbon dioxide from the alkali metal salts in aqueous solution. Acidifying agents which release carbon dioxide from the alkali metal salts in aqueous solution include, for example, boric acid and alkali metal hydrogen sulfates, alkali metal dihydrogen phosphates and other inorganic salts. However, preference is given to using organic acidifying agents, the citric acid being a particularly preferred acidifying agent. Acidifying agents in the effervescent system from the group of organic di-, tri- and oligocarboxylic acids or mixtures are preferred.

As perfume oils or perfumes, within the scope of the present invention, individual fragrance compounds, e.g. the synthetic products of the ester, ether, aldehyde, ketone, alcohol and hydrocarbon type are used. 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, e.g. Pine, citrus, jasmine, patchouly, rose or ylang-ylang oil.

In order to be perceptible, a fragrance must be volatile, whereby besides the nature of the functional groups and the structure of the chemical compound, the molecular weight also plays an important role. For example, most odorants have molecular weights up to about 200 daltons, while molecular weights of 300 daltons and above are more of an exception. Due to the different volatility of fragrances, the smell of a perfume or fragrance composed of several fragrances changes during evaporation, whereby the odor impressions in "top note", "middle note" or "body note" ) and "base note" (end note or dry out). Since odor perception is also largely due to odor intensity, the top note of a perfume does not consist solely of volatile compounds, while the base note is largely made up of less volatile, i. adherent fragrances. For example, in the composition of perfumes, more volatile fragrances can be bound to certain fixatives, preventing them from evaporating too quickly. In the subsequent classification of the fragrances in "more volatile" or "adherent" fragrances so nothing about the olfactory impression and whether the corresponding fragrance is perceived as a head or middle note, nothing said.

The fragrances can be processed directly, but it can also be advantageous to apply the fragrances on carriers that provide a slower fragrance release for long-lasting fragrance. As such carrier materials, for example, cyclodextrins have been proven, the cyclodextrin-perfume complexes can be additionally coated with other excipients. dyes

Preferred dyes, the selection of which presents no difficulty for 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 the substrates to be treated with the dye-containing agents, such as textiles, glass, ceramics or plastic crockery do not stain them.

When choosing the colorant, it must be taken into account that the colorants have a high storage stability and insensitivity to light. At the same time, it should also be taken into account when choosing suitable colorants that colorants have different stabilities to oxidation. In general, water-insoluble colorants are more stable to oxidation than water-soluble colorants. Depending on the solubility and thus also on the sensitivity to oxidation, the concentration of the colorant in the detergents or cleaners varies. In the case of readily water-soluble colorants, colorant concentrations in the range of a few 10 ^-2 to 10 ³ % by weight are typically selected. By contrast, in the case of the pigment dyes preferred in particular but less readily water-soluble on account of their brilliance, the suitable concentration of the colorant in detergents or cleaners is typically about 10 ³ to 10 ^-4 % by weight.

Dyeing agents which can be oxidatively destroyed in the washing process and mixtures thereof with suitable blue dyes, so-called blue toners, are preferred. It has proved to be advantageous to use colorants which are soluble in water or at room temperature in liquid organic substances. Suitable are, for example, anionic colorants, e.g. anionic nitrosofarads.

In addition to the components described in detail so far, the detergents and cleaners can contain further ingredients which further improve the performance and / or aesthetic properties of these compositions. Preferred agents contain one or more of the group of electrolytes, pH adjusters, fluorescers, hydrotopes, foam inhibitors, silicone oils, anti redeposition agents, optical brighteners, grayness inhibitors, anti-shrinkage agents, anti-crease agents, color transfer inhibitors, antimicrobial agents, germicides, fungicides, antioxidants, antistatic agents, ironing aids , Phobic and impregnating agents, swelling and anti-slip agents and UV absorbers.

As electrolytes from the group of inorganic salts, a wide number of different salts can be used. Preferred cations are the alkali and alkaline earth metals, preferred anions are the halides and sulfates. Out From a manufacturing point of view, the use of NaCl or MgCl ₂ in the washing or cleaning agents is preferred.

In order to bring the pH of detergents or cleaners into the desired range, the use of pH adjusters may be indicated. Can be used here are all known acids or alkalis, unless their use is not for technical application or environmental reasons or for reasons of consumer protection prohibited. Usually, the amount of these adjusting agents does not exceed 1% by weight of the total formulation.

Suitable foam inhibitors are, inter alia, soaps, oils, fats, paraffins or silicone oils, which may optionally be applied to support materials. Suitable carrier materials are, for example, inorganic salts such as carbonates or sulfates, cellulose derivatives or silicates and mixtures of the abovementioned materials. In the context of the present application preferred agents include paraffins, preferably unbranched paraffins (n-paraffins) and / or silicones, preferably linear-polymeric silicones, which are constructed according to the scheme (R ₂ SiO) X and are also referred to as silicone oils. These silicone oils usually are clear, colorless, neutral, odorless, hydrophobic liquids having a molecular weight between 1000 and 150,000, and viscosities between 10 and 1,000,000 mPa.s.

Suitable anti-redeposition agents, which are also referred to as soil repellents, are, for example, nonionic cellulose ethers such as methylcellulose and methylhydroxypropylcellulose with a proportion of methoxy groups of 15 to 30% by weight and of hydroxypropyl groups of 1 to 15% by weight, based in each case on the nonionic cellulose ether and the known from the prior art polymers of phthalic acid and / or terephthalic acid or derivatives thereof, in particular polymers of ethylene terephthalates and / or

Polyethylene glycol terephthalates or anionic and / or nonionic modified derivatives of these. Especially preferred of these are the sulfonated derivatives of the phthalic and terephthalic acid polymers.

Optical brighteners (so-called "whiteners") may be added to laundry detergents or cleaners to remove graying and yellowing of the treated fabrics which will attract the fiber and cause lightening and fake bleaching by exposing invisible ultraviolet radiation to visible, longer wavelength light convert the absorbed from sunlight ultraviolet light is radiated as a pale blue fluorescence and the yellow shade of the grayed or yellowed laundry to yield pure white Suitable compounds originate for example from the substance classes of 4.4. ^'- ^diamino-2,2' - stilbenedisulfonic (flavonic), ^{4,4'-biphenylene} -Distyryl, Methylumbelliferone, coumarins, dihydroquinolinones, 1, 3-diaryl pyrazolines, naphthalimides, benzoxazole, Benzisoxazole and benzimidazole systems and substituted by heterocycles pyrene derivatives.

Grayness inhibitors have the task of keeping the dirt detached from the fiber suspended in the liquor and thus preventing the dirt from being rebuilt. Water-soluble colloids of mostly organic nature are suitable for this purpose, for example the water-soluble salts of polymeric carboxylic acids, glue, gelatin, salts of ether sulfonic acids or 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, soluble starch preparations and other than the above-mentioned starch products can be used, e.g. degraded starch, aldehyde levels, etc. Also polyvinylpyrrolidone is useful. Cellulosic ethers such as carboxymethylcellulose (Na salt), methylcellulose, hydroxyalkylcellulose and mixed ethers such as methylhydroxyethylcellulose, methylhydroxypropylcellulose, methylcarboxymethylcellulose and mixtures thereof can furthermore be used as graying inhibitors.

Since textile fabrics, in particular of rayon, rayon, cotton and their mixtures, can tend to wrinkle because the individual fibers are sensitive to bending, buckling, pressing and crushing transversely to the fiber direction, synthetic anti-crease agents can be used. These include, for example, synthetic products based on fatty acids, fatty acid esters, fatty acid amides, alkylol esters, -alkylolamides or fatty alcohols, which are usually reacted with ethylene oxide, or products based on lecithin or modified phosphoric acid ester.

Phobic and impregnation processes are used to furnish textiles with substances that prevent the deposition of dirt or facilitate its leaching ability. Preferred repellents and impregnating agents are perfluorinated fatty acids, also in the form of their aluminum u. Zirconium salts, organic silicates, silicones, polyacrylic acid esters with perfluorinated alcohol component or polymerizable compounds coupled with perfluorinated acyl or sulfonyl radical. Antistatic agents may also be included. The antisoiling equipment with repellents and impregnating agents is often classified as an easy-care finish. The penetration of the impregnating agent in the form of solutions or emulsions of the active substances in question can be facilitated by adding wetting agents which reduce the surface tension. A further field of application of repellents and impregnating agents is the water-repellent finish of textiles, tents, tarpaulins, leather, etc., in which, in contrast to waterproofing, the fabric pores are not closed, so the fabric remains breathable (hydrophobing). The water repellents used for hydrophobizing coat textiles, leather, paper, wood, etc. with a very thin layer of hydrophobic groups, such as longer alkyl chains or siloxane groups. Suitable water repellents are, for example, paraffins, waxes, metal soaps, etc. with additions of aluminum or zirconium salts, quaternary Ammonium compounds with long-chain alkyl radicals, urea derivatives, fatty acid-modified melamine resins, chromium complex salts, silicones, tin organic compounds and glutaric dialdehyde and perfluorinated compounds. The hydrophobized materials do not feel greasy; nevertheless, similar to greasy substances, water droplets emit from them without moistening. For example, silicone-impregnated textiles have a soft feel and are water and dirt repellent; Stains from ink, wine, fruit juices and the like are easier to remove.

Antimicrobial agents can be used to combat microorganisms. Depending on the antimicrobial spectrum and mechanism of action, a distinction is made between bacteriostats and bactericides, fungistatics and fungicides, etc. Important substances from these groups are, for example, benzalkonium chlorides, alkylarylsulfonates, halophenols and phenolmercuric acetate, although it is entirely possible to do without these compounds.

To prevent undesirable changes to the detergents and cleaners and / or the treated fabrics caused by exposure to oxygen and other oxidative processes, the compositions may contain anti-oxidants. This class of compounds includes, for example, substituted phenols, hydroquinones, catechols and aromatic amines, as well as organic sulfides, polysulfides, dithiocarbamates, phosphites and phosphonates.

Increased comfort may result from the additional use of antistatic agents. Antistatic agents increase the surface conductivity and thus allow an improved drainage of formed charges. External antistatic agents are generally substances with at least one hydrophilic molecule ligand and give a more or less hygroscopic film on the surfaces. These mostly surface-active antistatic agents can be subdivided into nitrogen-containing (amines, amides, quaternary ammonium compounds), phosphorus-containing (phosphoric acid esters) and sulfur-containing (alkyl sulfonates, alkyl sulfates) antistatic agents. Lauryl (or stearyl) dimethylbenzylammonium chlorides are also suitable as antistatic agents for textiles or as an additive to detergents, wherein additionally a softening effect is achieved.

Silicone derivatives can be used to improve the water absorbency, rewettability of the treated fabrics, and ease of ironing the treated fabrics. These additionally improve the rinsing out of detergents or cleaning agents by their foam-inhibiting properties. Preferred silicone derivatives are, for example, polydialkyl or alkylaryl siloxanes in which the alkyl groups have one to five carbon atoms and are completely or partially fluorinated. Preferred silicones are polydimethylsiloxanes, which may optionally be derivatized and are then amino-functional or quaternized or have Si-OH, Si-H and / or Si-Cl bonds. Other preferred silicones are Polyalkylene oxide-modified polysiloxanes, ie polysiloxanes which comprise, for example, polyethylene glycols and the polyalkylene oxide-modified dimethylpolysiloxanes.

Finally, according to the invention, it is also possible to use UV absorbers which are absorbed by the treated textiles and improve the light resistance of the fibers. 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. Also suitable are substituted benzotriazoles, phenyl-substituted acrylates (cinnamic acid derivatives) in the 3-position, optionally with cyano groups in the 2-position, salicylates, organic Ni complexes and natural substances such as umbelliferone and the body's own urocanic acid.

Protein hydrolyzates are due to their fiber-care effect further in the context of the present invention preferred active substances from the field of detergents and cleaners. Protein hydrolysates are product mixtures obtained by acid, alkaline or enzymatically catalyzed degradation of proteins (proteins). According to the invention, protein hydrolysates of both vegetable and animal origin can be used. Animal protein hydrolysates are, for example, elastin, collagen, keratin, silk and milk protein hydrolysates, which may also be present in the form of salts. Preferred according to the invention is the use of protein hydrolysates of plant origin, e.g. Soy, almonds, rice, pea, potato and wheat protein hydrolysates. Although the use of the protein hydrolysates is preferred as such, amino acid mixtures or individual amino acids obtained otherwise, such as, for example, arginine, lysine, histidine or pyrroglutamic acid, may also be used in their place. Also possible is the use of derivatives of protein hydrolysates, for example in the form of their fatty acid condensation products.

Another object of the invention is a means for generating a light emission in a chemiluminescent assay (signal reagent) comprising an oxidase according to the invention, an oxidase substrate and a chemiluminescent reagent. The oxidase substrate is preferably different from choline. Preferably, the chemiluminescent reagent is luminol.

Chemiluminescent assays are of considerable importance in the field of medicine and the life sciences. Particularly important are immunoassays and DNA probe analyzes. Advantageously, oxidases of the present invention which generate oxygen-containing free radicals (eg, superoxide anion, hydroxyl radical) and peroxides (hydrogen peroxide) provide long-lived chemiluminescent detectable products upon reaction with chemiluminescent reagents such as lucigenin, luminol, and its derivatives. These oxidase systems are of particular use as so-called tracers for the detection of analytes in immunoassays, Immunoblotting or nucleotide probe analysis to provide long-lived light-emitting moieties after reaction with a chemiluminescent reagent.

Another embodiment of the invention comprises a kit containing an agent as described above. Chemiluminescent assays according to the invention are provided, for example, in sales units, so-called kits, which contain all the components necessary for carrying out the test, for example further reagents for a positive control, etc., as well as precise instructions for carrying out the test.

The following examples illustrate the invention without, however, limiting it to:

Example 1 :

Production of oxidases according to the invention by saturation mutagenesis of a choline oxidase as starting enzyme.

The A. nicotianae choline oxidase is present in the E. coli expression plasmid pET26b (+) of the company Novagen. The gene was amplified by polymerase chain reaction (PCR), according to the protocol of the "Herculase HotStart Polymerase" from Stratagene with the following oligonucleotides as primers:

a) "Forward_Ndel": δ ^{^{'-TTCCATATGAACATTGAAAAGAAGGACTT-S'}} b) 'Reverse_Xhol ": ^5' -CCGCTCGAGGGCTTCGCTAACCAGTTCGC-3 ^'

The amplification product obtained was inserted via the restriction sites Ndel and Xhol, so that the C-terminal histidine tag is used. The saturation mutagenesis of amino acid position E306 was performed by Stratagene's QuikChange Site-Directed Mutagenesis Kit, replacing the glutamate codon with codons for the 19 other proteinogenic amino acids of the modified enzymes using Novagen's "Overnight Express Autoinduction System", the cell lysates obtained by standard digestion using ultrasound were purified via nickel-NTA affinity chromatography and then examined for their biochemical characteristics. For this purpose, the cell lysates with Ni-NTA agarose from the company Qiagen were purified according to the protocol for the recovery of native proteins to the apparent homogeneity on the Coomassie-stained SDS-PAGE gel. Subsequently, among other things, the determination of the specific activity of the enzymes with various substrates. Example 2: Determination of Specific Activity

Calculation and definition:

The specific activity is obtained as a quotient of the activity and the protein content. The activity determination is based on the detection of hydrogen peroxide which is formed during the reaction catalyzed by the oxidase from the substrate and atmospheric oxygen. Here, the term one unit (E) (often also referred to as a "unit" (U)) refers to that amount of enzyme which produces one μmol of H ₂ O ₂ in one minute, indicating the catalytic activity concentration in units E / μl or mE / μl (corresponds to 10 ^{~ 3} U / μl) or as U / μl or mU / μl as activity per sample volume The protein concentration is determined with a commercially available test and expressed in mg / ml The standard curve is compared with bovine serum albumin (" bovine serum albumin ", BSA).

1. Solutions

To determine the oxidase activity, the following solutions are needed:

Peroxide reagent: 75 mM K ₂ HPO ₄ , 125 mM NaH ₂ PO ₄ , 12.5 mM chromotropic acid and 0.6 mM

aminoantipyrine

200 mM phosphate buffer pH = 6.5: 75 mM K ₂ HPO ₄ , 125 mM NaH ₂ PO ₄

Peroxidase solution: 52 U / mL horseradish peroxidase in phosphate buffer

Davis buffer: 25 mM citric acid x 1 H ₂ O, 25 mM NaB ₄ O ₇ x 1 H ₂ O, 25 mM KCl, 25 mM KH ₂ PO ₄ ,

25 mM tris- (hydroxymethyl) -aminomethane, pH 9.5

20 mM NaN ₃ in Davis buffer pH = 9.5

Substrate solution: 250 mM substrate in dist. H ₂ O. Substrates used: choline chloride, 3,3-

Dimethylbutanol, 3-methyl-1,3-butanediol, benzyl alcohol, 2-phenylethanol, isethionic acid (Na salt).

2nd implementation:

To 150 .mu.l of oxidase-containing solution (if necessary diluted accordingly with distilled H ₂ O) are added 30 .mu.l 20 mM NaN ₃ solution, 45 .mu.l Davis buffer pH = 9.5 and 150 .mu.L substrate solution and the mixture after mixing for Incubated for 30 min at 800 rpm and 37 ⁰ C. Subsequently, 525 μL peroxide reagent and 75 μL peroxidase solution are added and incubated at room temperature for 5 min. Finally, the absorbance of the solution is determined at a wavelength of λ = 600 nm.

The activity is determined by comparison with a calibration curve made with a suitable dilution series of hydrogen peroxide in 20 mM phosphate buffer (pH 6.5) under the conditions described above. The catalytic activity concentration in units (E) / μl is given as activity per sample volume.

The amount of protein per sample volume is determined with the BCA Protein Assay Kit (Pierce Biotechnology, Cat # 23227) according to the protocol proposed by the manufacturer and in mg / ml specified. The specific activity refers to the activity per mg protein and is given in E / mg or mE / mg = 10 ³ U / mg.

For comparison, the specific activity of the modified oxidase according to the invention with reference to a substrate other than choline is related to the specific activity of the enzyme with the reaction of choline. For this, the relative specific activities are formed as the quotients (hereinafter referred to as the substance coefficient) from the specific activity of the oxidase of the present invention with respect to a non-choline substrate and the choline-specific activity.

Example 3 Determination of the Substance Quotient and the Increase of the Mutant Quotient

The substance quotient is the quotient of the specific activity of an oxidase according to the invention on a substrate under consideration and the specific activity of precisely this oxidase on the original substrate choline. For example, if the specific activity of an oxidase according to the invention whose glutamic acid residue at position 306 has been changed (Glu306Xxx) is 10 U / mg on a new substrate (S), but only 5 U / mg on the substrate choline, then the corresponding substance ratio is 10 / 5 = 2. It can be used as a measure to check the selectivity of an oxidase according to the invention for a desired new substrate out.

Listed in Table 1 below is the amino acid at position 306 of the choline oxidase modified according to this position in accordance with SEQ ID NO: 1 in the first line.

Table 1: Representation of the substance quotient

The change of position 306 to Asp or Gly causes a very strong decrease of the undesired activity on choline with simultaneously lower decrease of the activity on desired substrates such as 3,3-dimethylbutanol, 3-methyl-1,3-butanediol, 2-phenylethanol and benzyl alcohol. The substrate specificity of the altered oxidases is therefore shifted away from choline to a non-choline substrate. The mutation to GIy is further characterized by the fact that it increases the activity on isethionic acid so that it is even higher than in the parent enzyme. At the same time, the activity on choline is greatly reduced.

The further changes Glu306 to GIn, Asn, Cys, Gly, Ala and Arg favor the change of choline to a new substrate, since the increase of the mutant quotient by 1 to 2 orders of magnitude higher than in the other investigated combinations. The mutant quotient is the ratio of the specific activity of an altered enzyme to the specific activity of the starting enzyme for an examined substrate. It thus indicates the extent to which a mutation has led to an improvement in specific activity relative to a particular, considered substrate or to what extent the specific activity of the starting enzyme relative to a substrate considered could be reduced by a change. The following calculation example for a fictitious substrate S serves to explain: The specific activity of the starting enzyme on S is 10 U / mg, whereas that of an altered enzyme Glu306Xxx is 15 U / mg on the substrate S. The mutant quotient is 15 U / mg divided by 10 U / mg = 1.5. By definition, the mutant quotient for the parent enzyme on all substrates = 1.

Listed in Table 2 below is the amino acid at position 306 of the choline oxidase modified according to this position in accordance with SEQ ID NO: 1 in the first line. The amino acid glutamic acid (Glu) originally present in the starting enzyme is marked as "wild-type" by "WT".

Table 2: Increase of the mutant quotient

A particularly desirable new substrate for new oxidases prepared in this way is 3-methyl-1, 3-butanediol, since it is favored by a particularly large number of different mutations, furthermore benzyl alcohol, followed by the other substrates investigated. Especially effective in the example considered is the change to isethionic acid in mutation to Cys or Gly and the change to benzyl alcohol by mutation to Cys. The mutation Glu306Cys for 3-methyl-1,3-butanediol, benzyl alcohol and 2-phenylethanol has particularly high selectivities for the new substrates investigated, since the substance quotients are greater than 1 here. The mutation Gly306Ala causes good selectivity for 3-methyl-1,3-butanediol.

Example 4:

In addition, according to the invention, oxidase enzymes can be produced and used in methods according to the invention whose activity towards a substrate other than choline is higher in comparison with the activity of the unaltered starting enzyme opposite this substrate. Thus, for example, an oxidase whose glutamic acid residue at position 306, based on SEQ ID NO: 1, has been replaced by a glycine residue (Glu 306 Gly) has an activity increased by the factor 1, 4 (the quotient of the activity of the modified enzyme Glu 306 Gly divided by the activity of the starting enzyme GIu 306 has a value of 1.4) with respect to the substrate isethionic acid compared with the starting enzyme, ie the oxidase with a glutamic acid residue at position 306. At the same time the activity on choline is greatly reduced (the quotient of the activity the altered enzyme Glu 306 Gly divided by the activity of the starting enzyme Glu 306 has a value of 0.01). Thus, according to the invention, it is also possible to obtain enzymes which not only exhibit a shift in substrate specificity towards non-choline substrates, but whose activity in relation to these substrates is also increased.

Description of the figure:

Figure 1: sequence comparison (alignment) of

N-terminal truncated choline oxidase from Arthrobacter nicotianae ("COD A.nicotianae shortened" SEQ ID NO: 1),

the choline oxidase from Arthrobacter nicotianae ("COD A.nicotianae", DSMZ-ID 96-878, accession number DSM 11234, SEQ ID NO: 3) and the

Arthrobacter globiformis choline oxidase ("COD A.globiformis"; SEQ ID NO: 2, database accession number AAP68832 of NCBI ("National Center for Biotechnology Information") protein database; UniProtKB / TrEMBL entry Q7X2H8) as published by Gadda et al , (Biochem Biophys., 421 (1), 149-158, 2004. "Cloning, sequence analysis, and purification of choline oxidase from Arthrobacter globiformis: a bacterial enzyme involved in osmotic stress tolerance").

From the sequence comparison, it is also clear that with regard to the method of counting the respective amino acid positions in the various polypeptide chains, for example in the truncated choline oxidase from Arthrobacter nicotianae ("COD A.nicotianae shortened", SEQ ID NO: 1), the missing amino acids must be taken into account accordingly ,

Claims

claims:

1. A process for the enzymatic production of hydrogen peroxide with an oxidase, characterized in that the amino acid sequence of the oxidase includes an amino acid sequence which is at least 96% identical to SEQ ID NO: 1, reacting a substrate which is different from choline.

A method according to claim 1, characterized in that the substrate other than choline is one having the structure R-CH 2 -OH, wherein R is 1-20 carbon atoms, 0-5 nitrogen atoms, 0-5 Oxygen atoms, 0-2 sulfur atoms, 0-2 phosphorus atoms and 0-10 halogen atoms.

A method according to claim 1 or 2, characterized in that the substrate other than choline is one having the structure R-CH 2 -OH, wherein R is selected from the group of -phenyl, -benzyl , -CH2-SO2-OH, -CH2-PO (OH) 2, -PO (OH) 2, -CH2-tert-butyl, -CH2-C (Me2, OH).

4. The method according to any one of claims 1 to 3, characterized in that it is the substrate, which is different from choline, 2-phenylethanol.

5. The method according to any one of claims 1 to 3, characterized in that it is the substrate, which is different from choline, 3-methyl-1, 3-butanediol.

6. The method according to any one of claims 1 to 3, characterized in that it is the substrate, which is different from choline, isethionic acid.

7. The method according to any one of claims 1 to 3, characterized in that it is the substrate, which is different from choline, is 3-benzyl alcohol.

8. The method according to any one of claims 1 to 7, characterized in that the substrate has a volume which is smaller than 2x 10 ^{~ 8} m ³ .

9. oxidase, characterized in that one or more amino acids at positions 55, 306, 325, 345, 351, 458, 459 and 460 are changed relative to SEQ ID NO: 1, wherein the amino acid at one of the positions 55, 325 , 345, 351, 458, 459 and 460 is selected from the Group of alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine and valine and differs from those described in SEQ ID NO: 1 is present at the corresponding position and the amino acid at position 306 is selected from the group of arginine, asparagine, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine , Threonine, tryptophan, tyrosine and valine.

10. oxidase according to claim 9, characterized in that it is more suitable for the oxidation of a substrate which is different from choline, as for the oxidation of choline.

11. oxidase according to claim 10, characterized in that it is the substrate other than choline, one of the structure R-CH2-OH, wherein R 1-20 carbon atoms, 0-5 nitrogen atoms, 0-5 Oxygen atoms, 0-2 sulfur atoms, 0-2 phosphorus atoms and 0-10 halogen atoms.

12. oxidase according to claim 10 or 11, characterized in that it is the substrate other than choline is one having the structure R-CH 2 -OH, wherein R is selected from the group of -phenyl, - benzyl , -CH2-SO2-OH, -CH2-PO (OH) 2, -PO (OH) 2, -CH2-tert-butyl, -CH2-C (Me2, OH).

13. oxidase according to any one of claims 10 to 12, characterized in that it is in the substrate, which is different from choline, 2-phenylethanol.

14. oxidase according to any one of claims 10 to 12, characterized in that it is in the substrate, which is different from choline, 3-methyl-1, 3-butanediol.

15. oxidase according to any one of claims 10 to 12, characterized in that it is the substrate, which is different from choline, isethionic acid.

16. oxidase according to any one of claims 10 to 12, characterized in that it is 3-benzyl alcohol in the substrate, which is different from choline.

17. oxidase according to any one of claims 10 to 16, characterized in that the substrate has a volume that is less than 2x 10 ^{~ 8} m ³ .

18. oxidase according to any one of claims 9 to 17, characterized in that the amino acid at position 306 is increasingly preferably arginine, glutamine, asparagine, aspartate or glycine, wherein the numbering of the sequence positions is based on SEQ ID NO: 1.

The oxidase of any one of claims 9 to 18, wherein the quotient formed from the specific activity of the oxidase using a substrate other than choline divided by the specific activity of the oxidase using a substrate which is choline, a value between 1 and 100,000, preferably a value between 1 and 1000, more preferably a value between 1 and 100 and particularly preferably a value between 1 and 10.

20. Nucleic acid molecule coding for an oxidase according to one of claims 9 to 18.

21. A vector containing a nucleic acid according to claim 20.

22. A vector according to claim 21, which is a cloning vector.

23. A vector according to claim 21, which is an expression vector.

24. A host cell which comprises an oxidase according to any one of claims 9 to 18 or a fragment thereof or which can be excited to produce them, preferably using an expression vector according to claim 23.

25. A host cell according to claim 24, characterized in that it secretes the oxidase or a fragment thereof in the medium surrounding the host cell.

26. A host cell according to claim 24 or 25, characterized in that it has a cell nucleus.

27. Host cell according to claim 24 or 25, characterized in that it is a bacterium.

28. A host cell according to claim 27, characterized in that the bacterium is selected from the group of the genera of Escherichia, Bacillus and Arthrobacter, Streptomyces and Pseudomonas.

29. A host cell according to claim 27 or 28, characterized in that the bacterium is selected from the group of Escherichia coli, Bacillus licheniformis, Bacillus amyloliquefaciens, Bacillus subtilis, Bacillus alcalophilus and Arthrobacter oxidans, Streptomyces lividans and Streptomyces coelicolor.

30. Use of an oxidase according to any one of claims 9 to 18 as a hydrogen peroxide generating agent.

31. Use of an oxidase according to any one of claims 9 to 18 for bleaching, for color transfer inhibition or for disinfection.

32. Composition containing an oxidase according to one of claims 9 to 18.

33. A composition according to claim 32, characterized in that it is a personal care product, shampoo, hair care products, oxidation, oral, dental or denture care, cosmetic, detergent, detergent, rinse aid, hand detergent, hand dishwashing detergent, machine dishwashing detergent, disinfectant, signal reagent or a means for bleaching or disinfecting treatment of filter media, textiles, furs, paper, skins or leather.

34. Composition according to one of claims 32 or 33, characterized in that it is present as a one-component system.

35. Composition according to one of claims 32 or 33, characterized in that it is divided into several components.

36. Composition according to one of claims 32 to 35, characterized in that it is present as free-flowing powder having a bulk density of 300 g / l to 1200 g / l, in particular 500 g / l to 900 g / l.

37. Composition according to one of claims 32 to 35, characterized in that it is present in pasty or liquid form.

38. A composition according to any one of claims 32 to 37, characterized in that the oxidase and / or its substrate is coated with a at room temperature or in the absence of water for the oxidase and / or its substrate impermeable substance.

39. Composition according to one of claims 32 to 38, further comprising

10% by weight to 65% by weight, in particular 12% by weight to 60% by weight of water-soluble or water-dispersible inorganic builder material, 0.5% by weight to 10% by weight, in particular 1% by weight to 8% by weight, of water-soluble organic builders,

0.01 to 15% by weight of solid inorganic and / or organic acids or acid salts,

0.01 to 5% by weight complexing agent for heavy metals,

0.01 to 5% by weight of grayness inhibitor,

0.01 to 5% by weight of color transfer inhibitor and

0.01 to 5% by weight foam inhibitor

40. Composition according to claim 39, further comprising from 0.01 to 5% by weight of optical brightener

41. Composition according to claim 39, further comprising from 0.01 to 5 wt .-% of one or more other enzymes.

42. A means for generating a light emission in a chemiluminescent assay comprising an oxidase according to any one of claims 9 to 18, further comprising an oxidase substrate and a chemiluminescent reagent.

43. The composition of claim 42, wherein the chemiluminescent reagent is luminol.

44. Kit containing agents according to claim 42 or 43.