EP2300587B1 - Washing and cleaning agent - Google Patents

Description

The present invention relates to enzyme-containing detergents or cleaners which, in addition to conventional constituents, contain combinations of chlorophyllases and galactolipases.

In addition to the ingredients indispensable for the washing process, such as surfactants and builders, detergents generally contain further constituents, which can be summarized by the term washing aids and which comprise such different active ingredient groups as foam regulators, grayness inhibitors, bleaching agents and color transfer inhibitors. Such adjuvants also include substances which aid surfactant performance by the enzymatic degradation of soil soils. The same applies mutatis mutandis to cleaners for hard surfaces. In addition to the protein removal assisting proteases and the fat-splitting lipases, the amylases, which have the task of facilitating the removal of starch-containing stains by the catalytic hydrolysis of the starch polysaccharide, and the cellulases are of particular importance.

Other common ingredients of detergents and cleaners are agents that are designed to remove stained soils such as tea or red wine stains. For this purpose, inorganic peroxygen compounds, especially hydrogen peroxide and solid peroxygen compounds which dissolve in water to release hydrogen peroxide, such as sodium perborate and sodium carbonate perhydrate, have long been used as oxidizing agents for disinfecting and bleaching purposes. The oxidation effect of these substances in dilute solutions depends strongly on the temperature. At lower temperatures, the oxidation effect of the inorganic peroxygen compounds can be improved by the addition of so-called bleach activators. Here come above all else Compounds from the classes of N- or O-acyl compounds, for example, polyacylated alkylenediamines or carboxylic acid esters in question. By adding these substances, the bleaching action of aqueous peroxide liquors can be increased so much that even at temperatures below 60 ° C substantially the same effects as with the peroxide solution alone at 95 ° C occur. Relevant type are US 2008/0070291 and WO 2006/099866 ,

Frequently, enzymes and bleaching systems complement each other in their effect, often even synergistic effects are observed. A particularly stubborn group of soils are chlorophyll-containing soils, especially grass or leaf stains, which can not be adequately removed by either the enzymes used or the bleach system. It proves to be problematic here that the chlorophyll is not combined in free form but in combination with hydrophobic or protein-containing plant constituents, whereby the attack of the bleaching systems on the chromophore is made more difficult.

Chlorophyllases (EC 3.1.1.14), which catalyze the cleavage of chlorophyll or pheophytin into chlorophyllide or pheophorbide and phythol as hydrolytic enzymes, have been known for about 100 years. The reaction of chlorophyllase with chlorophyll substantially improves the water solubility of the chromophore system.
Lipases are now routinely used in detergent formulations to remove lipid or grease stains. These enzymes remove the fatty contaminants by hydrolysis of one or more ester bonds of triacylglycerides, as well as phospholipids. A specific group of lipases are the galacto-lipases, which exclusively or in addition to triacylglycerides and phospholipids cleave one or more ester bonds of galactolipids.

The use of galactolipases in commercial detergent formulations has not been described previously. In galactolipids, one or more galactose residues are linked to the sn-3 position of diacylglycerides. Galactolipids are the main components of photosynthetically active membranes and are therefore found mainly in plants and photosynthetically active bacteria. In these galactolipid membranes, the chlorophyll molecules are embedded. Galactolipases are found, for example, in plants, where they occur mainly in the chloroplasts. Further sources of galactolipolytic enzymes are lipases from the mammalian digestive tract and these activities have also been demonstrated in microorganisms.

Surprisingly, it has now been found that the combination of chlorophyllases and galactolipases leads to unexpected synergistic performance improvements on chlorophyll-containing stains, so that this enzyme combination is particularly suitable for use in detergents and cleaners.

The invention relates to detergents or cleaners containing a combination of a chlorophyllase and a hydrolase which is a galactolipase.

By using this combination of a chlorophyllase and at least one other hydrolase, which is a galactolipase, the cleaning performance of detergents and cleaners is increased, especially toward colored, chlorophyll-based soiling, especially in aqueous washing and cleaning solutions containing a peroxygen compound. The term cleaning performance against colored stains is to be understood in its broadest meaning and includes both the bleaching of dirt located on the textile, the bleaching of befindlichem in the wash liquor, detached from the textile dirt as well as the oxidative destruction of themselves in the wash liquor Textile paints that detach under the washing conditions of textiles before they can be applied to differently colored textiles. Also when used in cleaning solutions for hard surfaces is under this term both the Bleaching understood on the hard surface befindendem dirt, especially tea, as well as the bleaching of befindlichem in the dishwashing liquor, detached from the hard surface dirt.

As one component, the washing or cleaning agent according to the invention contains a chlorophyllase. As chlorophyllases mainly vegetable enzymes are used, preferably enzymes from orange (Citrus sinensis) or from wheat (Triticum aestivum). For use in the products according to the invention, the enzymes can be produced recombinantly for example in Escherichia coli or Pichia pastoris and subsequently purified from the cytoplasmic crude extract or the culture supernatant by standard methods.

As a further constituent, the washing or cleaning agent according to the invention contains a galactolipase. The galactolipases used may on the one hand be of prokaryotic origin, for example from Pseudomomas sp. or Chromobacter sp. On the other hand, eukaryotic galactolipases from yeasts, fungi, as well as from plant or animal sources can be used, for example from Candida sp., Beans (Phaseolus vulgaris), potatoes (Solanum tuberosum) guinea pigs (Cavia porcellus), horse (Equus caballus) or human (Homo sapiens). For use in the products according to the invention, the enzymes can also be produced recombinantly, for example in Escherichia coli or Pichia pastoris, and purified from the cytoplasmic crude extract or the culture supernatant by standard methods.

A washing or cleaning agent according to the invention generally contains in each case 0.0001 to 10 mg, preferably in each case 0.001 mg to 1.0 mg, in particular in each case 0.02 to 0.3 mg of chlorophyllase and further hydrolase per gram of the washing and cleaning agent.

The detergents and cleaners according to the invention, which are in the form of pulverulent solids, in densified particle form, as homogeneous solutions or Suspensions may contain, in addition to the enzymes mentioned in principle all known and customary in such agents ingredients. The compositions of the invention may include surfactants, builders, bleaches based on organic and / or inorganic peroxygen compounds, additional bleach activators, bleach catalysts, water-miscible organic solvents, additional enzymes, sequestering agents, electrolytes, pH regulators, and other adjuvants such as optical brighteners, grayness inhibitors, dye transfer inhibitors , Foam regulators, silver corrosion inhibitors and dyes and fragrances. The compositions according to the invention may comprise one or more surfactants, in particular anionic surfactants, nonionic surfactants and mixtures thereof, but also cationic, zwitterionic and amphoteric surfactants.

Suitable nonionic surfactants are in particular alkyl glycosides and ethoxylation and / or propoxylation of alkyl glycosides or linear or branched alcohols each having 12 to 18 carbon atoms in the alkyl moiety and 3 to 20, preferably 4 to 10 alkyl ether groups. Also suitable are ethoxylation and / or propoxylation products of N-alkylamines, vicinal diols, fatty acid esters and fatty acid amides, which correspond to said long-chain alcohol derivatives with respect to the alkyl moiety, and of alkylphenols having from 5 to 12 carbon atoms in the alkyl radical.

The nonionic surfactants used are preferably alkoxylated, advantageously ethoxylated, in particular primary, alcohols having preferably 8 to 18 carbon atoms and on average 1 to 12 moles of ethylene oxide (EO) per mole of alcohol, in which the alcohol radical can be linear or preferably methyl-branched in the 2-position or may contain linear and methyl-branched radicals in the mixture, as they are usually present in Oxoalkoholresten. In particular, however, alcohol ethoxylates with linear radicals of alcohols of native origin having 12 to 18 carbon atoms, for. From coconut, palm, tallow or oleyl alcohol, and on average from 2 to 8 EO per mole of alcohol. Among the preferred ethoxylated alcohols include, for example, C ₁₂ -C ₁₄ -alcohols with 3 EO or 4 EO, C ₉ -C ₁₁ -alcohols with 7 EO, C ₁₃ -C ₁₅ -alcohols with 3 EO, 5 EO, 7 EO or 8 EO, C ₁₂ -C ₁₈ alcohols with 3 EO, 5 EO or 7 EO and mixtures of these, such as mixtures of C ₁₂ -C ₁₄ -alcohol with 3 EO and C ₁₂ -C ₁₈ -alcohol with 7 EO. The degrees of ethoxylation given represent statistical means which, for a particular product, may be an integer or a fractional number. Preferred alcohol ethoxylates have a narrow homolog distribution (narrow range ethoxylates, NRE). In addition to these nonionic surfactants, fatty alcohols with more than 12 EO can also be used. Examples include (tallow) fatty alcohols with 14 EO, 16 EO, 20 EO, 25 EO, 30 EO or 40 EO. In particular, in detergents for use in automatic dishwashing processes usually extremely low-foam compounds are used. These include preferably C ₁₂ -C ₁₈ -Alkylpolyethylenglykolpolypropylenglykolether each with at 8 mol ethylene oxide and propylene oxide in the molecule. However, it is also possible to use other known low-foam nonionic surfactants, such as, for example, C ₁₂ -C ₁₈ -alkyl polyethylene glycol-polybutylene glycol ethers having in each case up to 8 mol of ethylene oxide and butylene oxide units in the molecule and end-capped alkylpolyalkylene glycol mixed ethers.

Also particularly preferred are the hydroxyl-containing alkoxylated alcohols, so-called hydroxy mixed ethers. The nonionic surfactants also include 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 represents a glycose unit having 5 or 6 C atoms, preferably glucose. The degree of oligomerization x, which indicates the distribution of monoglycosides and oligoglycosides, is an arbitrary number - which, as a variable to be determined analytically, can also assume a fractional value - between 1 and 10; preferably x is 1.2 to 1.4. Also suitable are polyhydroxy fatty acid amides.

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

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.

Other suitable surfactants are so-called gemini surfactants. These are generally understood as meaning those compounds which have two hydrophilic groups per molecule. These groups are usually separated by a so-called "spacer". This spacer is typically a carbon chain that should be long enough for the hydrophilic groups to be spaced sufficiently apart for them to act independently of each other. In exceptional cases, the term gemini surfactants not only such "dimer", but also corresponding to "trimeric" surfactants understood. Suitable gemini surfactants are, for example, sulfated hydroxy mixed ethers or dimer alcohol bis and trimer alcohol tris sulfates and ether sulfates. End-capped dimeric and trimeric mixed ethers are characterized in particular by their bi- and multi-functionality. However, it is also possible to use gemini-polyhydroxy fatty acid amides or poly-polyhydroxy fatty acid amides.

Suitable anionic surfactants are in particular soaps and those which contain sulfate or sulfonate groups. As surfactants of the sulfonate type are preferably C ₉ -C ₁₃ - alkylbenzenesulfonates, olefinsulfonates, that is mixtures of alkene and hydroxyalkanesulfonates and disulfonates, as they are for example, from C ₁₂ -C ₁₈ monoolefins having terminal or irmenständiger double bond obtained by sulfonation with gaseous sulfur trioxide and subsequent alkaline or acidic hydrolysis of the sulfonation, into consideration. Also suitable are alkanesulfonates which are obtained from C ₁₂ -C ₁₈ -alkanes, for example by sulfochlorination or sulfoxidation with subsequent hydrolysis or neutralization. Also suitable are the esters of alpha-sulfo fatty acids (ester sulfonates), for example the alpha-sulfonated methyl esters of hydrogenated coconut, palm kernel or tallow fatty acids obtained by alpha-sulfonation of the methyl esters of fatty acids of plant and / or animal origin with 8 to 20 C -Atomen in the fatty acid molecule and subsequent neutralization to water-soluble mono-salts are prepared, into consideration. These are preferably the alpha-sulfonated esters of hydrogenated coconut, palm, palm kernel or tallow fatty acids, with sulfonation of unsaturated fatty acids, such as oleic acid, in small amounts, preferably in amounts not above about 2 to 3 wt. %, can be present. In particular, alpha-sulfofatty acid alkyl esters are preferred which have an alkyl chain with not more than 4 C atoms in the ester group, for example methyl ester, ethyl ester, propyl ester and butyl ester. With particular advantage, the methyl esters of the alpha-sulfo fatty acids (MES), but also their saponified disalts are used.

Other suitable anionic surfactants are sulfated fatty acid glycerol esters, which are mono-, di- and triesters and mixtures thereof, as in the preparation by esterification by a monoglycerol with 1 to 3 moles of fatty acid or in the transesterification of triglycerides with 0.3 to 2 moles of glycerol to be obtained. Examples of alk (en) ylsulfates are the alkali metal salts and, in particular, the sodium salts of the sulfuric monoesters 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 ₂₀ oxo alcohols and those half-esters of secondary alcohols of this chain length are preferred.

Also preferred are alk (en) ylsulfates of said chain length, which are a synthetic, petrochemical-based straight-chain Alkyl radical containing an analogous degradation behavior as the adequate compounds based on oleochemical raw materials. Of washing-technical interest, C ₁₂ -C ₁₆ -alkyl sulfates and C ₁₂ -C ₁₅ -alkyl sulfates and C ₁₄ -C ₁₅ -alkyl sulfates are particularly preferred. Also 2,3-alkyl sulfates are suitable anionic surfactants. Also suitable are the sulfuric acid monoesters of straight-chain or branched C ₇ -C ₂₁ -alcohols ethoxylated with from 1 to 6 mol of ethylene oxide, such as 2-methyl-branched C ₉ -C ₁₁ -alcohols having on average 3.5 mol of ethylene oxide (EO) or C ₁₂ - C ₁₈ -fatty alcohols with 1 to 4 EO. The preferred anionic surfactants also include the salts of alkylsulfosuccinic acid, which are also referred to as sulfosuccinates or as sulfosuccinic acid esters, and the monoesters and / or diesters of sulfosuccinic acid with alcohols, preferably fatty alcohols and in particular ethoxylated fatty alcohols. Preferred sulfosuccinates contain C ₈ to C ₁₈ 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.

Suitable further anionic surfactants are fatty acid derivatives of amino acids, for example N-methyltaurine (Tauride) and / or N-methylglycine (sarcosides). Particularly preferred are the sarcosides or the sarcosinates and here especially sarcosinates of higher and optionally monounsaturated or polyunsaturated fatty acids such as oleyl sarcosinate. As further anionic surfactants are particularly soaps into consideration. Particularly 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. Together with these soaps or as a substitute for soaps, it is also possible to use the known alkenylsuccinic acid salts.

The anionic surfactants, including 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.

Surfactants are present in detergents according to the invention in proportions of preferably from 5% by weight to 50% by weight, in particular from 8% by weight to 30% by weight, whereas agents for cleaning hard surfaces, in particular for machine cleaning of crockery , Lower surfactant levels of up to 10 wt .-%, in particular up to 5 wt .-%, and preferably in the range of 0.5 wt .-% to 3 wt .-% have.

An agent according to the invention preferably contains at least one water-soluble and / or water-insoluble, organic and / or inorganic builder. The water-soluble organic builders include polycarboxylic acids, especially citric acid and sugar acids; monomeric and polymeric aminopolycarboxylic acids, in particular methylglycinediacetic acid, nitrilotriacetic acid and ethylenediaminetetraacetic acid and polyaspartic acid, polyphosphonic acids, in particular aminotris (methylenephosphonic acid), ethylenediaminetetrakis (methylenephosphonic acid) and 1-hydroxyethane-1,1-diphosphonic acid, polymeric hydroxy compounds such as dextrin and also polymeric (poly) carboxylic acids, in particular the accessible by oxidation of polysaccharides or dextrins polycarboxylates, polymeric acrylic acids, methacrylic acids, maleic acids and copolymers thereof, which also small. Particles of polymerizable substances without carboxylic acid functionality may contain polymerized. The molecular weight of the homopolymers of unsaturated carboxylic acids is generally between 3000 and 200,000, and of the copolymers between 2000 and 200,000, preferably 30,000 to 120,000, in each case based on the free acid. A particularly preferred acrylic acid-maleic acid copolymer has a molecular weight of from 30,000 to 100,000 on. Commercially available products are, for example, Sokalan ™ CP 5, CP 10 and PA 30 from BASF. Suitable, although less preferred, compounds of this class are copolymers of acrylic or methacrylic acid with vinyl ethers, such as vinylmethyl ethers, vinyl esters, ethylene, propylene and styrene, in which the acid content is at least 50% by weight. It is also possible to use terpolymers which contain two unsaturated acids and / or salts thereof as monomers and also vinyl alcohol and / or an esterified vinyl alcohol or a carbohydrate as the third monomer as water-soluble organic builder substances. The first acidic monomer or its salt is derived from a monoethylenically unsaturated C ₃ -C ₈ -carboxylic acid and preferably from a C ₃ -C ₄ -monocarboxylic acid, in particular from (meth) -acrylic acid. The second acidic monomer or its salt may be a derivative of a C ₄ -C ₈ -dicarboxylic acid, with maleic acid being particularly preferred, and / or a derivative of an allylsulfonic acid which is substituted in the 2-position by an alkyl or aryl radical. Such polymers generally have a molecular weight between 1000 and 200,000. Further preferred copolymers are those which preferably have as monomers acrolein and acrylic acid / acrylic acid salts or vinyl acetate. The organic builder substances can be used, in particular for the preparation of liquid agents, in the form of aqueous solutions, preferably in the form of 30 to 50 percent by weight aqueous solutions. All of the acids mentioned are generally used in the form of their water-soluble salts, in particular their alkali metal salts.

If desired, such organic builder substances may be present in amounts of up to 40% by weight, in particular up to 25% by weight and preferably from 1% by weight to 8% by weight. Quantities close to the stated upper limit are preferably used in paste-form or liquid, in particular water-containing, agents according to the invention.

Suitable water-soluble inorganic builder materials are, in particular, alkali metal silicates, alkali metal carbonates and alkali metal phosphates, which may be in the form of their alkaline, neutral or acidic sodium or potassium salts. Examples of these are trisodium phosphate, tetrasodium diphosphate, disodium dihydrogen diphosphate, pentasodium triphosphate, so-called sodium hexametaphosphate, oligomeric trisodium phosphate with degrees of oligomerization of from 5 to 1000, in particular from 5 to 50, and the corresponding potassium salts or mixtures of sodium and potassium salts. Crystalline or amorphous alkali metal aluminosilicates, in amounts of up to 50% by weight, preferably not more than 40% by weight, and in liquid agents, in particular from 1% by weight to 5% by weight, are particularly suitable as water-insoluble, water-dispersible inorganic builder materials. used. Among these, the crystalline sodium aluminosilicates in detergent quality, more particularly zeolite A, P and optionally X, alone or in mixtures, for example in the form of a co-crystals of zeolites A and X (VEGOBOND ^® TM AX, a commercial product of Condea Augusta SpA), prefers. Amounts near the above upper limit are preferably used in solid, particulate agents. In particular, suitable aluminosilicates have no particles with a particle size greater than 30 μm and preferably consist of at least 80% by weight of particles having a size of less than 10 μm. Their calcium binding capacity is usually in the range of 100 to 200 mg CaO per gram.

Suitable substitutes or partial substitutes for the said aluminosilicate are crystalline alkali silicates which may be present alone or in a mixture with amorphous silicates. The alkali metal silicates useful as builders in the compositions according to the invention preferably have a molar ratio of alkali oxide to SiO ₂ below 0.95, in particular from 1: 1.1 to 1:12, and may be present in amorphous or crystalline form. Preferred alkali metal silicates are the sodium silicates, in particular the amorphous sodium silicates, with a molar ratio of Na 2 O: SiO 2 of from 1: 2 to 1: 2.8. The crystalline silicates which may be present alone or in admixture with amorphous silicates, are crystalline layer silicates with the general formula Na ₂ SIXO ₂ x + 1.y H ₂ O used in which x, the so-called module, a number from 1.9 to 22, in particular 1.9 to 4 and y is a number from 0 to 33 and preferred values for x are 2, 3 or 4. Preferred crystalline layered silicates are those in which x in the aforementioned general Formula takes 2 or 3. In particular, _(y H ₂ O Na ₂ Si ₂ O _5.) Are both beta and delta-sodium preferred. Also prepared from amorphous alkali silicates, practically anhydrous crystalline alkali silicates of the abovementioned general formula in which x is a number from 1.9 to 2.1, can be used in inventive compositions. In a further preferred embodiment of the composition according to the invention, a crystalline sodium layer silicate with a modulus of 2 to 3 is used. Crystalline sodium silicates with a modulus in the range from 1.9 to 3.5 are used in a further preferred embodiment of compositions according to the invention. Crystalline layered silicates of the above formula (I) are sold by Clariant under the tradename Na-SKS, e.g. Na-SKS-1 (Na ₂ Si ₂₂ O ₄₅ .xH ₂ O, Kenyaite), Na-SKS-2 (Na ₂ Si ₁₄ O ₂₉ .xH ₂ O, magadiite), Na-SKS-3 (Na ₂ Si ₈ O ₁₇ .xH ₂ O) or Na-SKS-4 (Na ₂ Si ₄ O ₉ .xH ₂ O, makatite). Of these, especially Na-SKS-5 (alpha-Na ₂ Si ₂ O ₅ ), Na-SKS-7 (beta-Na ₂ Si ₂ O ₅ , natrosilite), Na-SKS-9 (Na ₂ Si ₂ O ₅ .3H ₂ O), Na-SKS-10 (NaHSi ₂ O ₅ .3H 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 (delta-Na ₂ Si ₂ O ₅ ). In a preferred embodiment of inventive compositions is a granular compound of crystalline layered silicate and citrate, of crystalline layered silicate and the above-mentioned (co) polymeric polycarboxylic acid or alkali metal silicate and alkali metal carbonate, as available for example under the name Nablon ^® ™ 15 commercially is.

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

Suitable peroxygen compounds are, in particular, hydrogen peroxide and inorganic salts which release hydrogen peroxide under the washing conditions, which include the alkali perborates, percarbonates, persilicates and / or persulphates such as caroate, but also organic peracids or persalts of organic acids such as phthalimidopercaproic acid, perbenzoic acid or salts the diperdodecanedioic acid, into consideration. If solid peroxygen compounds are to be used, they can be used in the form of powders or granules, which can also be enveloped in a manner known in principle. Peroxygen compounds are present in amounts of preferably up to 50% by weight, more preferably from 5% to 30% and most preferably from 8% to 25% by weight. The addition of small amounts of known bleach stabilizers such as phosphonates, borates or metaborates and metasilicates and magnesium salts such as magnesium sulfate may be useful.

As bleach activators it is possible, in particular, 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, 2,5-diacetoxy- 2,5-dihydrofuran and enol ester, as well as acetylated sorbitol and Mannitol, acylated sugar derivatives, in particular pentaacetylglucose (PAG), pentaacetyl fructose, tetraacetylxylose and octaacetyl lactose as well as acetylated, optionally N-alkylated glucamine and gluconolactone, and / or N-acylated lactams, for example N-benzoylcaprolactam. Hydrophilic substituted acyl acetals and acyl lactams are also preferably used.

Such bleach activators may be present in the customary amount range, preferably in amounts of from 0.5% by weight to 10% by weight, in particular from 1% by weight to 8% by weight, based on the total agent. In addition to the conventional bleach activators listed above or in their place, sulfone imines and / or bleach-enhancing transition metal salts or transition metal complexes can also be present as so-called bleach catalysts. Among the candidate transition metal compounds include in particular manganese, iron, cobalt, ruthenium or molybdenum-salene complexes and their known N-analogues, manganese, iron, cobalt, ruthenium or molybdenum carbonyl complexes, manganese, iron , Cobalt, ruthenium, molybdenum, titanium, vanadium and copper complexes with nitrogenous tripod ligands, cobalt, iron, copper and ruthenium ammine complexes. Bleach-enhancing transition metal complexes, in particular having the central atoms Mn, Fe, Co, Cu, Mo, V, Ti and / or Ru, are used in customary amounts, preferably in an amount of up to 1% by weight, in particular 0.0025% by weight. % to 0.25 wt .-% and particularly preferably from 0.01 wt .-% to 0.1 wt .-%, each based on the total agent used.

Furthermore, detergents and cleaners according to the invention may contain proteases, further lipases, amylases and / or cellulases. Suitable proteases include those of microorganisms, especially bacteria or fungi, recoverable enzymes with a pH optimum in the alkaline range. Protease is preferably used in the agent according to the invention in amounts such that the finished composition is 100 PE / g to 7500 PE / g (protease units per gram, determined by the method described in Tenside 7, 125 (1970)), in particular 125 PE / g to 5000 PE / g and more preferably 150 PE / g to 4500 PE / g. Useful proteases are in the Commercially available, for example, under the names BLAP ™, Savinase ™, Esperase ™, Maxatase ™, Optimase ™, Alcalase ™, Durazym ™, Everlase ™, Maxapem® &, or Purafect ™ OxP.

The amylases which can be used in the compositions according to the invention, which are preferably used in combination with at least one further enzyme, include the enzymes which can be obtained from bacteria or fungi and which have a pH optimum preferably in the alkaline range up to about pH 10. Useful commercial products include Termamyl ™, Maxamyl ™, Duramyl ™, or Purafect ™ OxAm. Amylase is preferably used in the composition according to the invention in amounts such that the finished product is 0.01 KNU / g to 2 KNU / g ("kilo-novo units" per gram according to the Novo standard method, with 1 KNU the amount of enzyme which degrades 5.26 g of starch at pH 5.6 and 37 ° C based on the method described by P. Bemfeld in SP Colowick and ND Kaplan, Methods in Enzymology, Vol. 1, 1955, page 149), especially 0.015 KNU / g up to 1.8 KNU / g and more preferably 0.03 KNU / g to 1.6 KNU / g. If the agent according to the invention contains an amylase, it is preferably selected from the genetically modified amylases.

The additional lipase optionally additionally present in the agent according to the invention is an enzyme which can be obtained from microorganisms, in particular bacteria or fungi. Lipase is preferably employed in the composition of the invention in amounts such that the finished composition has a lipolytic activity in the range of 10 LU / g to 10,000 LU / g ("lipase activity units" per gram as determined by the enzymatic hydrolysis of tributyrin at 30 ° C and pH 7 according to the method mentioned in EP 258 068), in particular 80 LU / g to 5000 LU / g and more preferably 100 LU / g to 1000 LU / g. Commercially available lipases are, for example, Lipolase ™, Lipomax ™, Lumafast ™ and Lipozym ™.

Likewise, the cellulase useful in the invention belongs to the enzymes obtainable from bacteria or fungi, which preferably have a pH optimum in the near neutral to slightly alkaline pH range from 6 to 9.5. They are preferably used in the composition according to the invention in amounts such that the finished composition has a cellulolytic activity of from 0.05 IU / g to 1.5 IU / g ("International Units" per gram, based on the enzymatic hydrolysis of Na-carboxymethylcellulose) pH 9.0 and 40 ° C, as in Agric. Biol. Chem. 53, 1275 (1989) by S. Ito et al. in particular 0.07 IU / g to 1.4 IU / g and more preferably 0.1 IU / g to 1.3 IU / g. Suitable commercial products are, for example, Celluzyme ™ from the manufacturer Novo Nordisk or KAC ™ from Kao.

Since a plurality of enzymes are to be used in the composition according to the invention, this can be carried out by incorporating the two or more separate or in a known manner separately prepared enzymes or by two or more together in a granule ready-made enzymes.

Suitable enzymes which can additionally be used in the compositions are those from the class of cutinases, pullulanases, hemicellulases, oxidases, laccases and peroxidases and mixtures thereof. Particularly suitable are from fungi or bacteria, such as Bacillus subtilis, Bacillus licheniformis, Bacillus lentus, Streptomyces griseus, Humicola lanuginosa, Humicola insolens, Pseudomonas pseudoalcaligenes, Pseudomonas cepacia or Coprinus cinereus derived enzymatic agents. The enzymes may be adsorbed to carriers and / or embedded in encapsulants to protect against premature inactivation. They are preferably present in the detergents or cleaners according to the invention in amounts of up to 5% by weight, in particular from 0.2% by weight to 4% by weight.

Among the agents according to the invention, in particular when they are in liquid or pasty form, organic solvents which can be used in addition to water, include alcohols having 1 to 4 C atoms, in particular methanol, ethanol, isopropanol and tert-butanol, diols having 2 to 4 C -Atomen, in particular ethylene glycol and propylene glycol, and mixtures thereof and from the mentioned compound classes derivable ether. Such water-miscible solvents are preferably present in the compositions according to the invention in amounts of not more than 30% by weight, in particular from 6% by weight to 20% by weight.

In addition, the agents may contain other ingredients customary in detergents and cleaners. These optional ingredients include, in particular, enzyme stabilizers, grayness inhibitors, color transfer inhibitors, foam inhibitors, and optical brighteners, as well as dyes and fragrances. To effect silver corrosion protection, silver corrosion inhibitors can be used in dishwashing detergents according to the invention. In addition, a hard surface cleaning agent according to the invention may contain abrasive constituents, in particular from the group comprising quartz flours, wood flours, plastic flours, chalks and glass microspheres and mixtures thereof. Abrasive substances are preferably not more than 20% by weight, in particular from 5% by weight to 15% by weight, in the cleaning agents according to the invention.

In order to establish a desired pH, which does not result from the mixture of the other components, the compositions according to the invention can contain system- and environmentally compatible acids, in particular citric acid, acetic acid, tartaric acid, malic acid, lactic acid, glycolic acid, succinic acid, glutaric acid and / or adipic acid. but also mineral acids, in particular sulfuric acid, or bases, in particular ammonium or alkali metal hydroxides. Such pH regulators are present in the compositions according to the invention in amounts of preferably not more than 20% by weight, in particular from 1.2% by weight to 17% by weight.

The color transfer inhibitors which are suitable for use in textiles according to the invention include in particular polyvinylpyrrolidones, polyvinylimidazoles, polymeric N-oxides such as poly (vinylpyridine-N-oxide) and copolymers of vinylpyrrolidone with vinylimidazole. Graying inhibitors have the task of keeping suspended from the textile fiber dirt suspended in the fleet. Water-soluble colloids of mostly organic nature are suitable for this purpose, for example starch, glue, gelatin, salts of ether carboxylic acids or ether sulfonic acids of starch or of cellulose or salts of acidic sulfuric acid esters of cellulose or starch. Also, water-soluble polyamides containing acidic groups are suitable for this purpose. Furthermore, other than the above-mentioned starch derivatives can be used, for example aldehyde starches. Preference is given to using cellulose ethers, such as carboxymethylcellulose (Na salt), methylcellulose, hydroxyalkylcellulose and mixed ethers, such as methylhydroxyethylcellulose, methylhydroxypropylcellulose, methylcarboxymethylcellulose and mixtures thereof, for example in amounts of from 0.1 to 5% by weight, based on the compositions.

To enhance the cleaning performance, in particular the detergents according to the invention may comprise soil-release polymers, which are generally composed of carboxylic acid units and optionally polymeric diol units and contain, for example, ethylene terephthalate and polyoxyethylene terephthalate groups. Other monomer units, for example, propylene glycol, polypropylene glycol, alkylene or alkenylene dicarboxylic acids, isophthalic acid, carboxy- or sulfo-substituted phthalic acid isomers may be included in the soil release polymer. End-capped derivatives, that is, polymers having neither free hydroxyl groups nor free carboxyl groups, but, for example C ₁ - ₄ alkyl groups or carry with monobasic carboxylic acids, for example benzoic acid or sulfobenzoic acid, esterified terminal, can be used. Also suitable are polyesters which, in addition to oxyethylene groups and terephthalic acid units, contain 1,2-propylene, 1,2-butylene and / or 3-methoxy-1,2-propylene groups and also glycerol units and are end-capped with C ₁ - to C ₄ -alkyl groups , the ethylene-terephthalate and polyethylene oxide terephthalate soil-release polymers having a molecular weight of 900 to 9000, wherein the polyethylene glycol units have molecular weights of 300 to 3000 and the molar ratio of ethylene terephthalate to Polyethylene terephthalate is from 0.6 to 0.95, at least in part by C ₁ - ₄ alkyl or acyl groups end-capped polyester with polypropylene terephthalate and polyoxyethylene terephthalate units, the sulfoethylendgruppenverschlossenen terephthalate-containing soil release polyester prepared by sulfonation of unsaturated end groups Soil-release polyester with terephthalate-, alkylene glycol- and poly-C ₂ - ₄ - glycol units cationic soil release polyester with amine, ammonium and / or amine oxide groups and the cationic soil release polyester with ethoxylated quaternized morpholine units. Also suitable are polymers of ethylene terephthalate and polyethylene terephthalate in which the polyethylene glycol units have molecular weights of from 750 to 5,000 and the molar ratio of ethylene terephthalate to polyethylene oxide terephthalate is from 50:50 to 90:10, and polymers of molecular weight from 15,000 to 50,000 of ethylene terephthalate and polyethylene oxide terephthalate, wherein the polyethylene glycol units have molecular weights of 1,000 to 10,000 and the molar ratio of ethylene terephthalate to polyethylene oxide terephthalate is 2: 1 to 6: 1.

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

In particular, when used in mechanical processes, it may be advantageous to add conventional foam inhibitors to the compositions. As foam inhibitors are, for example, soaps of natural or synthetic origin, which have a high proportion of C ₁₈ -C ₂₄ fatty acids. Suitable non-surfactant Foam inhibitors are, for example, organopolysiloxanes and mixtures thereof with microfine, optionally silanized silica and paraffins, waxes, microcrystalline waxes and mixtures thereof with silanated silicic acid or bis-fatty acid alkylenediamides. It is also advantageous to use mixtures of various foam inhibitors, for example those of silicones, paraffins or waxes. The foam inhibitors, in particular silicone- and / or paraffin-containing foam inhibitors, are preferably bound to a granular, water-soluble or dispersible carrier substance. In particular, mixtures of paraffins and bistearylethylenediamide are preferred.

The production of solid compositions according to the invention can be carried out in a known manner, for example by spray-drying or granulation, the enzymes and any further thermally sensitive ingredients such as, for example, bleaching agents optionally being added separately later. For the preparation of compositions according to the invention having an increased bulk density, in particular in the range from 650 g / l to 950 g / l, a known process comprising an extrusion step is preferred. Another preferred preparation by means of a granulation process is in the European patent specification EP 0 642 576 described.

For the preparation of compositions according to the invention in tablet form, which may be monophasic or multiphase, monochromatic or multicolor and in particular consist of one or more layers, in particular two layers, the procedure is preferably such that all constituents - if appropriate one per layer - in one Mixer mixed together and the mixture by means of conventional tablet presses, such as eccentric or rotary presses pressed. Particularly in the case of multilayer tablets, it can be advantageous if at least one layer is pre-compressed. Preferably, a tablet produced in this way has a weight of from 10 g to 50 g, in particular from 15 g to 40 g. The spatial form of the tablets is arbitrary and can be round, oval or angular, with intermediate forms are also possible. Corners and edges are advantageously rounded. Point round tablets preferably a diameter of 30 mm to 40 mm. In particular, the size of rectangular or cuboid-shaped tablets, which are introduced predominantly via the metering device, for example the dishwasher, is dependent on the geometry and the volume of this metering device. Exemplary preferred embodiments have a base area of (20 to 30 mm) x (34 to 40 mm), in particular of 26 x 36 mm or 24 x 38 mm.

Liquid or pasty detergents or cleaners according to the invention in the form of conventional solvent-containing solutions are generally prepared by simply mixing the ingredients, which can be added in bulk or as a solution in an automatic mixer.

The preparation of chlorophyllase according to the invention by cloning is known from the literature (IntEnz Enzyme Nomenclature EC 3.1.1.1.14), z. B. according to Tsuchiya et al., Proc. Natl. Acad. Sci. USA 96, 15362-15367 (1999 ). Examples of chlorophyllases are known under Arabidopsis thaliana AT1G19670 or AT5G43860. Galactolipases are known under EC 3.1.1.26, CAS registry number: 37278-40-3.

Examples 1. Expression of a recombinant chlorophyllase in E. coli

The chlorophyllase gene was amplified by the polymerase chain reaction from a cDNA sample with the oligonucleotides Citrus_CHL_fwd_Ndel and Citrus_CHL_rev_Xhol as primers. For directed cloning into a corresponding expression vector, for example pET28a (Novagen), recognition sequences for restriction endonucleases (Ndel and Xhol) could be incorporated simultaneously into the oligonucleotides. The vector pET28a contained the bacteriophage T7 promoter system and encoded a C-terminal and / or N-terminal His-tag. The amplified DNA was mixed with the digested with both restriction endonucleases and fractionated and purified on an agarose gel, with the appropriate band being excised from the gel and extracted. The digested and gel-purified PCR product was ligated with the cut with the same restriction endonucleases and dephosphorylated expression vector. The ligated DNA could subsequently be used for the transformation of electrocompetent E. coli DH10B (Invitrogen). Positive transformants could be identified by colony PCR, restriction analysis and sequencing.

For expression of the chlorophyllase, E. coli tuner (DE3) pLacI (Novagen) could be transformed with a plasmid clone containing a verified insert. Cultures were grown in 2xYT medium supplemented with kanamycin (50 μg / ml) and chloramphenicol (34 μg / ml) at 37 ° C. At an optical density of about 0.8, expression was induced by the addition of 1mM isopropyl thiogalactoside (IPTG). Subsequently, the cultivation was continued at 30 ° C for four to six hours.

2. Purification of a Recombinant Chlorophyllase from E. coli

The cells were harvested by centrifugation and resuspended in 20 mM sodium phosphate buffer (pH 7.4) with 500 mM NaCl, 20 mM imidazole and 0.5 mg / ml lysozyme and incubated at room temperature. The cell disruption was carried out by freezing three times in liquid nitrogen with subsequent thawing at about 42 ° C or by ultrasound. The batch was centrifuged with the chlorophyllase as a soluble protein in the supernatant.

The chlorophyllase was then further purified by metal affinity chromatography. For this purpose, the metal affinity matrix was equilibrated with 20 mM sodium phosphate buffer (pH 8) with 500 mM NaCl, 20 mM imidazole and 10% glycerol, after sample application the matrix was washed with said buffer and then the chlorophyllase with 20 mM sodium phosphate buffer ( pH 8) eluted with 500 mM NaCl, 250 mM imidazole and 10% glycerol.

3. Measuring chlorophyllase activity

The activity of the expressed chlorophyllase was determined by the increased water solubility of the resulting reaction product chlorophyllide. The reaction was carried out in a 100 μl batch containing 100 μM chlorophyll of spinach (Fluka), 20% acetone (v / v), and 100 mM Na-MOPS pH 7.0. The reaction mixture was incubated for 60 min at 37 ° C. with shaking and then stopped by addition of 50 μl acetone, 50 μl n-hexane and 5 μl Tris-Cl (2 M, pH 9.0). The mixture was well homogenized by vigorous shaking and then the phase separation was accelerated by centrifugation for two minutes. By the reaction of chlorophyllase, the water-soluble chlorophyllide was in the lower aqueous and the unreacted chlorophyll in the upper organic phase. For quantification, 80 μl of the aqueous phase were admixed with 120 μl of methanol and the fluorescence of the excited chlorophyllide was measured (Ex 355 nm, Em 660 nm). The amount of chlorophyllide formed was determined using a standard calibration curve.

4. Expression of a recombinant galactolipase in Pichia pastoris

The synthetic gene cassette of human pancreatic lipase related protein 2, a galactolipase flanked by recognition sites for restriction endonucleases Kpnl and Pagl, was cloned into a Pichia expression vector such as pGK1 or pGAPZα. For this purpose, both the synthetic gene and the corresponding expression vector, which was subsequently dephosphorylated, digested with the restriction endonucleases according to the manufacturer's instructions. The digested nucleic acids were size fractionated and purified by agarose gel electrophoresis and then ligated with T4 DNA ligase according to the manufacturer's instructions. The ligated DNA was then used for the transformation of electrocompetent E. coli DH10B. Positive transformants were identified by colony PCR, restriction analysis and sequencing.

For expression of the lipase Pichia pastoris was transformed with a verified plasmid clone. The expression was on YPD plates, spiked with Tributyrin, detected due to halo formation with active lipase secretion. From lipase-active clones, liquid cultures were subsequently added in YPD medium mixed with zeocin (100 μg / l). The cultures were incubated for 48 to 72 hours at 30 ° C with shaking. Thereafter, the cells were separated from the culture supernatant in which the lipase was located by centrifugation. The culture supernatant was desalted by diafiltration and concentrated by lyophilization.

5. Measuring lipase activity

The activity of the lipase was monitored by increasing the absorbance at 405 nm during the hydrolysis of p-nitrophenyl butyrate pNP-C4 or p-nitrophenyl caprylate pNP-C8. The reaction was carried out in a 1 ml batch containing 2 mM of the p-nitrophenyl ester, 50 mM potassium phosphate buffer (pH 8) and 0.1% Triton X-100. The increase in absorbance at 405 nm was monitored continuously over a period of at least one minute. With the aid of the extinction coefficient of p-nitrophenol ε _{(pNP, 405 nm, pH 8)} = 16.05 mM ^-1 cm ^-1 , the volume activity was determined from the increase. One unit corresponds to the amount of enzyme which catalyzes the release of 1 μmol of p-nitrophenol in one minute in the test batch described above at 22 ° C.

6. Determination of chlorophyllide release from chloroplasts

Chloroplasts for this assay were isolated from spinach leaves. For this, fresh spinach leaves were ground with sea sand in a mortar and homogenized with 50 mM potassium phosphate buffer pH 8 and 0.33 M sucrose in a mortar. The suspension was filtered through eight layers of cheesecloth and the debris and remaining sand were removed by centrifugation at 200xg for one minute. The chloroplasts in the supernatant were then pelleted by centrifugation at 1000xg for 10 minutes. The chloroplast pellet was resuspended in 50 mM HEPES pH 7.6 with 2 mM EDTA, 1 mM MgCl ₂ and 0.33 M sorbitol. The chlorophyll content is determined by measuring the absorbance at 652 nm in 80% acetone with ε = 34.5 lg ^-1 cm ^-1 .

Chlorophyllide release from chloroplasts was determined by the increased water solubility of the reaction product. Chloroplasts corresponding to a chlorophyll content of 10 μg were dissolved in a 150 μl reaction mixture containing 50 mM potassium phosphate buffer pH 8 with different amounts of chlorophyllase (eg 250 ng) and / or lipase (eg 5 U pNP-). C4) for 60 min at 40 ° C. Subsequently, the chloroplasts were pelleted and the liberated chlorophyllide determined in the supernatant. The determination of chlorophyllide was carried out by measuring the absorbance at 652 nm or by measuring the fluorescence in 60% MeOH (Em 355 nm, Ex 655 nm).

Results: Control (without enzymes) Chlorophyllase (Example 2) Galactolipase (example 4) Chlorophyllase / galatolipase Released chlorophyllide in the supernatant 1.7% 3.7% 14.7% 25.2%

7. Removal of grass stains from textile surfaces.

To determine the detergency cotton fabric in Läppchenform (WFK 10A, Laundry Research Krefeld) was evenly dyed with freshly cut grass. For aging, the stains were stored dry for at least 3 days with exclusion of light.

Subsequently, the lobules (about 1 cm ² ) were first treated for 2 h at 37 ° C with 500 ul of enzyme solution containing ~3 ug chlorophyllase (Example 2) and ~7.5 U (pNP-C4) galactolipase (Example 4). The lobules were then washed with a detergent solution (Spee Color detergent, Henkel Dusseldorf) at 40 ° C by hand. The degree of removal of the chlorophyll-containing stain was then determined with a whiteness meter. The delta R value is shown as the difference between the treated and the untreated test tissue.

Washing results (% remission)

Tissue washed with Spee color solution 36.9% remission Spee Color solution plus chlorophyllase 44.6% remission Spee Color solution plus chlorophyllase plus galactolipase 54.5% remission

It can be seen that the inventive combination of a chlorophyllase with a hydrolase has a significantly improved cleaning performance, as the use of a chlorophyllase without the addition of a hydrolase.

Claims (5)

1. A washing and cleaning composition comprising a chlorophyllase and a further hydrolase which is a galactolipase.
2. The washing and cleaning composition as claimed in claim 1, which comprises 0.0001 mg to 10 mg of chlorophyllase and 0.0001 mg to 10 mg of hydroiase per gram.
3. The washing and cleaning composition as claimed in claim 1, which additionally comprises a protease.
4. The washing and cleaning composition as claimed in claim 1, which additionally comprises an amylase.
5. The use of a washing and cleaning composition as claimed in one or more of claims 1 to 4 for removing chlorophyll-containing stains.