WO2009021867A2 - Agents contenant des protéases - Google Patents

Agents contenant des protéases Download PDF

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Publication number
WO2009021867A2
WO2009021867A2 PCT/EP2008/060204 EP2008060204W WO2009021867A2 WO 2009021867 A2 WO2009021867 A2 WO 2009021867A2 EP 2008060204 W EP2008060204 W EP 2008060204W WO 2009021867 A2 WO2009021867 A2 WO 2009021867A2
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WIPO (PCT)
Prior art keywords
protease
amino acid
acid sequence
seq
proteases
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PCT/EP2008/060204
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German (de)
English (en)
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WO2009021867A3 (fr
WO2009021867A9 (fr
Inventor
Cornelius Bessler
Arnd Kessler
Susanne Tondera
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Henkel Ag & Co. Kgaa
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Publication of WO2009021867A2 publication Critical patent/WO2009021867A2/fr
Publication of WO2009021867A3 publication Critical patent/WO2009021867A3/fr
Publication of WO2009021867A9 publication Critical patent/WO2009021867A9/fr

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
    • C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
    • C12N9/52—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from bacteria or Archaea
    • C12N9/54—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from bacteria or Archaea bacteria being Bacillus
    • C—CHEMISTRY; METALLURGY
    • C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
    • C11D—DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
    • C11D3/00—Other compounding ingredients of detergent compositions covered in group C11D1/00
    • C11D3/16—Organic compounds
    • C11D3/38—Products with no well-defined composition, e.g. natural products
    • C11D3/386—Preparations containing enzymes, e.g. protease or amylase

Definitions

  • the invention relates to middle compositions, in particular detergents and cleaners, containing a first protease and a second protease. Furthermore, the invention relates to purification processes in which these agents are used, uses of these agents and purification processes and uses of these proteases.
  • proteases of the subtilisin type are particularly important, which are attributed to the serine proteases due to the catalytically active amino acids. They act as nonspecific endopeptidases, that is, they hydrolyze any acid amide linkages that are internal to peptides or proteins. Their pH optimum is usually in the clearly alkaline range.
  • Subtilases become natural formed by microorganisms; Of these, in particular, the subtilisins formed and secreted by Bacillus species are to be mentioned as the most important group within the subtilases.
  • Proteases are, in addition to other enzymes, established active ingredients of detergents and cleaners. They cause the breakdown of protein-containing stains on the items to be cleaned. At best, there are synergies between the enzymes and the remaining components of the funds concerned.
  • the detergent and detergent proteases subtilases occupy an outstanding position due to their favorable enzymatic properties such as stability or pH optimum. They are also suitable for a variety of other technical uses, for example as components of cosmetics or in the organic-chemical synthesis.
  • subtilisin-type proteases preferably used in detergents and cleaners are the subtilisins BPN 'and Carlsberg, the protease PB92, the subtilisins 147 and 309, the alkaline protease from Bacillus lentus, subtilisin DY and those the subtilases, but no longer Enzymes Thermitase, Proteinase K, and Proteases TW3 and TW7, which can be classified as subtilisins in the narrower sense.
  • subtilisin BPN ' which is derived from Bacillus amyloliquefaciens, or B. subtilis, is known from the work of Vasantha et al. (1984) in J. Bacteriol., Volume 159, pp. 811-819 and JA Wells et al. (1983) in Nucleic Acids Research, Volume 11, pp. 7911-7925.
  • Subtilisin BPN ' is used in particular with regard to the numbering of the positions as reference enzyme of Subtilisins.
  • Subtilisin Carlsberg is available in a further developed form under the trade name Alcalase® from Novozymes A / S, Bagsvaerd, Denmark. It is described in the publications of EL Smith et al.
  • protease PB92 is naturally derived from the alkaliphilic bacterium Bacillus nov. spec. 92 and was available under the trade name Maxacal® from Gist-Brocades, Delft, The Netherlands. In its original sequence, it is described in the patent application EP 283075 A2.
  • the subtilisins 147 and 309 are sold under the trade names Esperase®, and Savinase® by the company Novozymes.
  • Bacillus strains which are disclosed in the application GB 1243784 A.
  • the protease from Bacillus lentus DSM 5483 (WO 91/02792 A1) is derived from the variants described under the name BLAP®, which are described in particular in WO 92/21760 A1, WO 95/23221 A1, WO 02/088340 A2 and WO 03 / 038082 A2.
  • Subtilisin DY is originally from Nedkov et al. Chem., 1985, Biol. Chem. Hoppe-Seyler, Vol. 366, pp. 421-430.
  • proteases are, for example, those under the trade names Durazym®, Relase®, Everlase®, Nafizym, Natalase®, Kannase® and Ovozyme® from Novozymes, which are available under the trade names, Purafect®, Purafect® OxP, Purafect® Prime and Properase ® from Genencor, sold under the trade name Protosol® by Advanced Biochemicals Ltd., Thane, India, under the trade name Wuxi® by Wuxi Snyder Bioproducts Ltd., China, under the trade names Proleather® and Protease P From Amano Pharmaceuticals Ltd., Nagoya, Japan, and that available under the name Proteinase K-16 from Kao Corp., Tokyo, Japan.
  • proteases used in the compositions according to the invention are either originally derived from microorganisms, for example from microorganisms of the genera Bacillus, Streptomyces, Humicola or Pseudomonas, and / or are produced by biotechnological methods known per se by suitable microorganisms, for example by transgenic expression hosts of the genera Bacillus or by filamentous fungi.
  • proteases can be used to improve the washing or cleaning performance together with other enzymes, for example amylases, cellulases, hemicellulases, mannanases, ⁇ -glucosidases, oxidases, oxidoreductases or lipases.
  • enzymes for example amylases, cellulases, hemicellulases, mannanases, ⁇ -glucosidases, oxidases, oxidoreductases or lipases.
  • proteases in detergents in combination with other active ingredients such as bleaching agents or soil release agents is known in the art.
  • corresponding agents in particular washing and cleaning agents, may contain a plurality of proteases.
  • agents in particular detergents and cleaners
  • the performance of agents is significantly improved with regard to their proteolytic activity, if mixtures of at least two specific proteases with different activity spectrum and / or different sequence are used in these agents.
  • different proteases results in a synergistic effect, ie a better performance compared to the individual performances of the respective protease in one-component systems (ie agents that contain only each of these proteases) and also to the sum of Individual performances of the proteases, ie the sum of, for example, two one-component systems.
  • the selected combination of particular proteases is another way to improve the performance of agents, especially detergents, with regard to their proteolytic activity. It is therefore an object of the present invention to provide agents which have improved proteolytic activity.
  • another object of the present invention is to provide means having improved detergency performance relative to at least one proteinaceous soiling, preferably with respect to multiple proteinaceous soils.
  • Another particular object of the present invention is to provide agents which have at least a constant, preferably improved, washing or cleaning performance with respect to at least one proteinaceous soiling, preferably with respect to a plurality of proteinaceous soils, but a reduced content Protease enzyme have.
  • the proteases contained develop a synergistic cleaning performance, so that the agents effect a better removal of proteinaceous soils compared to agents containing either only one protease type or compared to the expected cleaning performance of a two-protease agent due to mere addition the jewiligen individual contributions of the contained proteases to the cleaning performance of the agent.
  • the selected combination of such proteases constitutes an essential aspect of the invention which has a synergistic effect on the removal of proteinaceous residues or soils in compositions of the invention.
  • An object of the invention thus represent means comprising a first protease and a second protease, which are characterized in that the first protease is selected from a) a protease comprising an amino acid sequence corresponding to the amino acid sequence given in SEQ ID NO.1 at least 80% identical; b) a protease comprising an amino acid sequence which is at least 80% identical to the amino acid sequence given in SEQ ID NO. 2; c) a protease which comprises an amino acid sequence which is at least 80% identical to the amino acid sequence given in SEQ ID NO. 2 and additionally has at least one additional amino acid in comparison with SEQ ID NO.
  • the second protease comprises an amino acid sequence which is at least 80% identical to the amino acid sequence given in SEQ ID NO.3. It has surprisingly been found that in particular the combination of a protease, as defined above under a) to c) (hereinafter collectively referred to as "BLAP-type protease” or as the first protease) with a protease corresponding to the one described in SEQ ID NO.3 indicated amino acid sequence is at least 80% identical (hereinafter referred to as protease of the type "alkaline protease from Bacillus gibsonii (DSM 14391)" or second protease), in an agent according to the invention leads to a synergistic cleaning performance of the two proteases.
  • the synergistic effect of these two types of proteases in an agent according to the invention is based on the fact that the effects of the first and the second protease complement one another in such a way that an overall increased proteolytic effect is achieved.
  • This can be made possible, for example, by the fact that the proteolytic cleavage of peptide bonds by the first protease makes substrates for the second protease more accessible and, in turn, further substrates for the first protease become accessible by the proteolytic cleavage of the peptide bonds by the second protease.
  • the accessibility of a cleavage site for a protease is often a limiting factor.
  • a different spectrum of action of the two proteases therefore shows, for example, in a different substrate specificity.
  • a different spectrum of action may be caused by a different proteolytic activity under defined conditions, in particular with regard to temperature, pH, ionic strength, stability towards oxidizing compounds (for example bleaching agents), etc.
  • a synergism according to the invention therefore occurs when the first protease differs from the second protease. These differences manifest themselves in the amino acid sequence of the respective proteolytically active enzyme.
  • the first protease it is preferably selected from
  • a protease which comprises an amino acid sequence which is more preferably at least 82.5%, 85%, 87.5%, 90%, 91%, 92%, 93% to the amino acid sequence given in SEQ ID NO.1, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% and most preferably 100% identical.
  • a protease comprising an amino acid sequence which is more preferably at least 82.5%, 85%, 87.5%, 90%, 91%, 92%, 93%, of the amino acid sequence given in SEQ ID NO. 94%, 95%, 96%, 97%, 98%, 99%, 99.5% and most preferably 100% identical.
  • a protease comprising an amino acid sequence which is more preferably at least 82.5%, 85%, 87.5%, 90%, 91%, 92%, 93% to the amino acid sequence given in SEQ ID NO. Is 94%, 95%, 96%, 97%, 98%, 99%, 99.5% and most preferably 100% identical and has at least one additional amino acid compared to SEQ ID NO.2.
  • the agent is characterized in that the additional amino acid of the first protease is inserted after one of the following positions in the counting manner according to SEQ ID NO.2: 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 95, 96, 97, 98, 99, 100, 101, 102, 103, 125, 126, 127, 126, 129, 130, 131, 132, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 202, 203, 204, 218, 219.
  • proteases which are used according to the invention as the first protease in an agent, have a corresponding insertion of at least one amino acid or of a plurality of amino acids, preferably at one or more of said positions on.
  • proteases which are described in the European patent application EP 1 032 655. These have, for example, at least one additional amino acid in the counting method according to SEQ ID NO. 2 (that is to say in the counting method of Savinase®, referred to in the named application as BLSAVI), in positions 33 to 43, 95 to 103, 125 to 132 , 153 to 173, 181 to 195, 202 to 204 and / or position 218 to 219.
  • proteases which have an insertion of at least one amino acid and can be used according to the invention as the first protease, are derived from the international patent application WO 00/37599 and the international patent application WO 01/44452.
  • the agent is characterized in that the second protease comprises an amino acid sequence which is more preferably at least 82.5% to the amino acid sequence given in SEQ ID NO. 85%, 87.5%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5%, and most preferably 100% identical is.
  • a synergistic proteolytic performance of a composition according to the invention is found, in particular, when a first protease of the BLAP type is combined with a second protease of the type "alkaline protease from Bacillus gibsonii (DSM 14391)."
  • a first protease of the BLAP type is combined with a second protease of the type "alkaline protease from Bacillus gibsonii (DSM 14391).
  • DSM 14391 Bacillus gibsonii
  • an enzyme is to be understood as meaning a protein which has a specific biocatalytic function.
  • protease is understood as meaning an enzyme which catalyzes the hydrolysis of peptide bonds and is thereby able to cleave peptides or proteins.
  • a protein is a largely linear composition composed of the natural amino acids and usually performs one of its functions three-dimensional structure accepting polypeptide.
  • a peptide consists of amino acids that are covalently linked to each other via peptide bonds.
  • 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.
  • the proteinogenic, naturally occurring L-amino acids are designated by the internationally used 1- and 3-letter codes.
  • pre-proteins ie together with a signal peptide.
  • 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.
  • the signal peptide is cleaved under natural conditions by a signal peptidase from the rest of the protein, 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.
  • the mature, ie mature, peptides, ie the enzymes processed after their preparation are preferred over the preproteins.
  • the SEQ ID NO. 1, SEQ ID NO. 2 and SEQ ID NO. The sequences can therefore be modified from the cells producing them after the 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.
  • 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.
  • alignments are created using computer programs, such as the algorithms FASTA or BLAST; This procedure is described, for example, by DJ Lipman and WR Pearson (1985) in Science, Vol. 227, pp. 1435-1441.
  • sequence comparisons and determinations of homology and / or identity values were performed using the computer program Vector NTI ® Suite 7.0, available from InforMax, Inc., Bethesda, USA with the preset default parameters.
  • 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 represented in percent identity, that is the proportion of identical nucleotides or amino acid residues at the same or in an alignment corresponding 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.
  • Proteases or enzymes in general can be prepared by various methods, e.g. targeted genetic modification by mutagenesis, further developed and optimized for specific uses or specific properties such as catalytic activity, stability, etc.
  • 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.
  • 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.
  • the purpose of such recombination may be to induce or modify a particular enzymatic function using the fused protein portion.
  • it is irrelevant whether such a chimeric protein consists of a single polypeptide chain or several subunits on which different functions can be distributed.
  • 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 from those only in the size ratio of the unchanged protein part to the size of the entire protein. In such insertionsmut elected proteins, the proportion of foreign protein is lower than in chimeric proteins.
  • Inversion mutagenesis ie a partial sequence reversal
  • 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.
  • ком ⁇ онентs 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.
  • a compound may, for example, also be other proteins which are bound, for example via bifunctional chemical compounds, to proteins according to the invention.
  • 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.
  • derivatization is understood to mean covalent attachment to a macromolecular carrier, as well as noncovalent inclusion in suitable macromolecular cage structures.
  • the agent is thus characterized in that the first protease is present in the agent as fragment, deletion variant, chimeric protein or derivative and / or the second protease is present in the agent as a fragment, deletion variant, chimeric protein or derivative, wherein the first and second proteases are still catalytically active.
  • compositions according to the invention include all types of agents, in particular mixtures, formulations, solutions, etc., whose applicability is improved by addition of the proteases described above.
  • these may be, for example, solid mixtures, for example powders with freeze-dried or encapsulated proteins, or gel or liquid agents.
  • Preferred formulations contain, for example, buffer substances, stabilizers, reaction partners and / or cofactors of the proteases and / or other ingredients synergistic with the proteases. In particular, this appropriation is to be understood as the areas of application set out below. Further fields of application emerge from the prior art and are described, for example, in the manual "Industrial Enzymes and their Applications" by H.
  • an agent according to the invention is characterized in that it comprises a detergent, hand washing detergent, dishwashing detergent, hand dishwashing detergent, machine dishwashing detergent, cleaning agent, denture or contact lens care agent, rinse aid, disinfectant, cosmetic agent, pharmaceutical agent or a means for treating filter media, textiles , Furs, paper, skins or leather, especially a laundry detergent or dishwashing detergent.
  • This invention includes all conceivable types of detergents or cleaners, both concentrates and undiluted agents to be used on a commercial scale, in the washing machine or in hand washing or cleaning.
  • detergents or cleaners include, for example, detergents for textiles, carpets, or natural fibers, for which according to the present invention the term laundry detergent is used.
  • laundry detergent includes, 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.
  • An agent according to the invention can be either a means for large consumers or technical users as well as a product for the private consumer, wherein all types of detergents and cleaning agents established in the prior art also constitute embodiments of the present invention.
  • the detergents or cleaning agents according to the invention may contain, in addition to the active ingredient used according to the invention - a protease combination according to the invention - in principle all known and conventional ingredients in such agents, wherein at least another ingredient is present in the agent.
  • the agents according to the invention may in particular be builders, surface-active surfactants, bleaches based on organic and / or inorganic peroxygen compounds, bleach activators, water-miscible organic solvents, enzymes, sequestering agents, electrolytes, pH regulators and other auxiliaries such as optical brighteners, grayness inhibitors, foam regulators and dyes and fragrances and combinations thereof.
  • a combination of the protease combination with one or more other ingredients of the compositions proves to be advantageous since such an agent has an improved cleaning performance by resulting synergisms, in particular between a protease or the protease combination and the further ingredient.
  • the agent provides improved removal of stains, such as proteinaceous Stains caused in comparison with an agent which either contains only one of the two components, or even compared to the expected cleaning performance of an agent with both components due to the mere addition of the respective individual contributions of these two components to the cleaning performance of the agent.
  • Such a synergism is achieved, in particular, by the combination of a protease or protease combination according to the invention with one of the surfactants and / or builders and / or bleaches described below.
  • 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. Furthermore, corresponding ethoxylation and / or propoxylation of N-alkyl-amines, 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 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.
  • EO ethylene oxide
  • 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 EO per mole of alcohol are preferred.
  • Preferred ethoxylated alcohols include, for example, Ci 2 - C 4 alcohols containing 3 EO or 4 EO, C 9 -C i-alcohols containing 7 EO, C 3 -C 5 alcohols containing 3 EO, 5 EO, 7 EO or 8 EO, Ci 2 -Ci 8 alcohols with 3 EO, 5 EO or 7 EO and mixtures of these, such as mixtures of Ci 2 -Ci 4 alcohol with 3 EO and Ci 2 -Ci 8 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 rank ethoxylates, NRE).
  • fatty alcohols with more than 12 EO can also be used. Examples of these are (TaIg) fatty alcohols with 14 EO, 16 EO, 20 EO, 25 EO, 30 EO or 40 EO.
  • agents for use in mechanical processes usually extremely low-foam compounds are used. These preferably include C 2 -C 8 -Alkylpolyethylenglykol- polypropylene glycol ethers, each containing up to 8 moles of ethylene oxide and propylene oxide units in the molecule.
  • low-foam nonionic surfactants such as, for example, C 1 -C 6 -alkylpolyethyleneglycol-polybutylene glycol ethers having up to 8 mol of ethylene oxide and butylene oxide units in the molecule and end-capped alkylpolyalkylene glycol mixed ethers.
  • C 1 -C 6 -alkylpolyethyleneglycol-polybutylene glycol ethers having up to 8 mol of ethylene oxide and butylene oxide units in the molecule and end-capped alkylpolyalkylene glycol mixed ethers.
  • hydroxyl-containing alkoxylated alcohols as described in European Patent Application EP 0 300 305, 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, may also assume fractional values - between 1 and 10; preferably x is 1, 2 to 1, 4.
  • polyhydroxy fatty acid amides of the formula (III) in which R 1 CO is an aliphatic acyl radical having 6 to 22 carbon atoms, R 2 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 preferably derived from reducing sugars having 5 or 6 carbon atoms, in particular from glucose.
  • the group of polyhydroxy fatty acid amides also includes compounds of the formula (IV)
  • R 3 is a linear or branched alkyl or alkenyl radical having 7 to 12 carbon atoms
  • R 4 is a linear, branched or cyclic alkylene radical or an arylene radical having 2 to 8 carbon atoms
  • R 5 is a linear, branched or cyclic alkyl radical or a Aryl radical or an oxy-alkyl radical having 1 to 8 carbon atoms, wherein dC 4 alkyl or phenyl radicals are preferred
  • [Z] is a linear polyhydroxyalkyl radical whose alkyl chain is substituted with at least two hydroxyl groups, or alkoxylated, preferably ethoxylated or propoxylated derivatives this rest stands.
  • [Z] is also here preferably by reductive amination of a sugar such as glucose, fructose, maltose, lactose, galactose, mannose or Get xylose.
  • a sugar such as glucose, fructose, maltose, lactose, galactose, mannose or Get xylose.
  • the N-alkoxy- or N-aryloxy-substituted compounds can then be converted into the desired polyhydroxy fatty acid amides, for example by reaction with fatty acid methyl esters in the presence of an alkoxide as catalyst.
  • 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.
  • nonionic 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. Such surfactants are generally characterized by an unusually low critical micelle concentration and the ability to greatly reduce the surface tension of the water. 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.
  • the end-capped surfactants mentioned have good wetting properties and are low foaming, so that they are particularly suitable for use in machine washing or cleaning processes.
  • gemini-polyhydroxy fatty acid amides or poly-polyhydroxy fatty acid amides it is also possible to use gemini-polyhydroxy fatty acid amides or poly-polyhydroxy fatty acid amides.
  • Schwefelkladoester the ethoxylated with 1 to 6 moles of ethylene oxide, linear or branched C 7 -C 2 i-alcohols such as 2-methyl-branched C 9 -CN alcohols containing on average 3.5 mol ethylene oxide (EO) or C 2 - Ci 8 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.
  • 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 particularly important prefers.
  • 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).
  • sarcosides or the sarcosinates and here especially sarcosinates of higher and optionally monounsaturated or polyunsaturated fatty acids such as oleyl sarcosinate.
  • 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 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 inventive compositions in proportions of preferably 5 wt .-% to 50 wt .-%, in particular from 8 wt .-% to 30 wt .-%.
  • 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 builder substances include polycarboxylic acids, in particular citric acid and sugar acids, monomeric and polymeric aminopolycarboxylic acids, in particular methylglycine diacetic 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 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 amounts of polymerizable substances without carboxylic acid functionality
  • the molecular weight of the homopolymers of unsaturated carboxylic acids is generally between 3,000 and 200,000, of the copolymers between 2,000 and 200,000, preferably 30,000 to 120,000, each based on the free acid.
  • a particularly preferred acrylic acid-maleic acid copolymer has a molecular weight of from 30,000 to 100,000.
  • 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 the Acrylic acid or methacrylic acid with vinyl ethers, such as vinylmethyl ethers, vinyl esters, ethylene, propylene and styrene, in which the proportion of the acid is at least 50% by weight.
  • 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 substituted in the 2-position with an alkyl or aryl radical.
  • Such polymers generally have a molecular weight between 1,000 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.
  • 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.
  • 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, tetra sodium 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.
  • suitable aluminosilicates have no particles with a particle size greater than 30 .mu.m and preferably consist of at least 80% by weight of particles having a size of less than 10 .mu.m.
  • Your calcium binding capacity which can be determined according to the specifications of German Patent DE 24 12 837, is generally 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 metal oxide to SiO 2 of less than 0.95, in particular of 1: 1, 1 to 1: 12, and may be amorphous or crystalline.
  • 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 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 Si x O y are used 2x + 1 H 2 O, in which x, known as the modulus, an integer of 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 phyllosilicates are those in which x in the abovementioned general formula assumes the values 2 or 3. In particular, both ⁇ - and ⁇ -sodium disilicates (Na 2 Si 2 O 5 y H 2 O) are preferred.
  • 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.
  • a crystalline sodium layer silicate with a modulus of 2 to 3 is used, as can be prepared from sand and soda. Crystalline sodium silicates with a modulus in the range of 1.9 to 3.5 are used in a further preferred embodiment of compositions according to the invention.
  • Crystalline layer-form silicates of formula (I) given above are sold by Clariant GmbH under the trade name Na-SKS, eg 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).
  • Na-SKS eg 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
  • Na-SKS-4 Na 2 Si 4 O 9 XH 2 O, makatite
  • Na-SKS-5 OC-Na 2 Si 2 O 5
  • Na-SKS-7 ⁇ -Na 2 Si 2 0 5 , natrosilite
  • Na-SKS-9 NaHSi 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 0 5
  • Na-SKS-13 NaHSi 2 O 5
  • Na-SKS-6 5-Na 2 Si 2 O 5 .
  • composition according to the invention a granular compound of crystalline phyllosilicate and citrate, of crystalline phyllosilicate and of the above-mentioned (co-) polymeric polycarboxylic acid, or of alkali silicate and alkali metal carbonate, such as, for example, commercially available under the name Nabion® 15, is used ,
  • Builder substances are preferably present in the compositions according to the invention in amounts of up to 75% by weight, in particular 5% by weight to 50.
  • Suitable peroxygen compounds for use in compositions according to the invention are, in particular, organic peracids or persistent salts of organic acids, such as phthalimidopercaproic acid, perbenzoic acid or salts of diperdodecanedioic acid, hydrogen peroxide and under the washing conditions hydrogen peroxide-releasing inorganic salts, which include perborate, percarbonate, persilicate and / or persulfate such as caroate into consideration.
  • organic peracids or persistent salts of organic acids such as phthalimidopercaproic acid, perbenzoic acid or salts of diperdodecanedioic acid, hydrogen peroxide and under the washing conditions hydrogen peroxide-releasing inorganic salts, which include perborate, percarbonate, persilicate and / or persulfate such as caroate into consideration.
  • solid peroxygen compounds are to be used, they can be used in the form of powders or granules, which can also be
  • an agent according to the invention contains peroxygen compounds, they are present in amounts of preferably up to 50% by weight, in particular from 5% by weight to 30% by weight.
  • bleach stabilizers such as phosphonates, borates or metaborates and metasilicates and magnesium salts such as magnesium sulfate may be useful.
  • 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.
  • 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 esters
  • TAED
  • hydrophilic substituted acyl acetals and the acyl lactams are also preferably used.
  • Combinations of conventional bleach activators can also be used.
  • Such bleach activators can, in particular in the presence of the abovementioned hydrogen peroxide-supplied bleach, in the usual amount range, preferably in amounts of from 0.5 wt .-% to 10 wt .-%, in particular 1 wt .-% to 8 wt .-%, based on However, total agent, be included, missing when using percarboxylic acid as the sole bleach, preferably completely.
  • 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 -Ato- men, in particular ethylene glycol and propylene glycol, and mixtures thereof and derived from the said classes of compounds ethers.
  • 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.
  • the compositions according to the invention may contain system and environmentally acceptable 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.
  • 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.
  • water-soluble polyamides containing acidic groups are suitable for this purpose.
  • starch derivatives can be used, for example aldehyde starches.
  • 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 ,
  • Detergents according to the invention may contain, for example, derivatives of diaminostilbenedisulfonic acid or their alkali metal salts as optical brighteners, although they are preferably free of optical brighteners for use as color detergents.
  • optical brighteners for use as color detergents.
  • 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 are used instead of the morpholino Group carry a diethanolamino group, a methylamino group, an anilino group or a 2-methoxyethylamino group.
  • brighteners of the substituted diphenylstyrene type may be present, for example, the alkali salts of 4,4'-bis (2-sulfostyryl) -diphenyl, 4,4'-bis (4-chloro-3-sulfostyryl) -diphenyl, or 4 - (4-chlorostyryl) -4 '- (2-sulfostyryl).
  • Mixtures of the aforementioned optical brightener can be used.
  • 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 bound to a granular, water-soluble or dispersible carrier substance.
  • mixtures of paraffins and bistearylethylenediamide are preferred.
  • the proteases are combined, for example, with one or more of the following ingredients: nonionic, anionic and / or cationic surfactants, (optionally further) bleaches, bleach activators, bleach catalysts, builders and / or cobuilders, Acids, alkaline substances, hydrotropes, solvents, thickeners, sequestering agents, electrolytes, optical brighteners, grayness inhibitors, corrosion inhibitors, in particular silver protectants (silver corrosion inhibitors), disintegration aids, soil release agents, color transfer (or transfer) inhibitors, foam inhibitors, abrasives, Dyes, fragrances, perfumes, antimicrobial agents, UV protectants or absorbents, antistatic agents, pearlescers and skin protection agents, enzymes such as protease, amylase, cellulase, hemicellulase, mannanase, tannase, xylanase, xanthine
  • nonionic, anionic and / or cationic surfactants
  • an agent according to the invention is therefore characterized in that it contains at least one further component selected from the group consisting of surfactants, builders, acids, alkaline substances, hydrotropes, solvents, thickeners, bleaching agents, dyes, perfumes, corrosion inhibitors , Sequestering agents, electrolytes, optical brighteners, grayness inhibitors, silver corrosion inhibitors, color transfer inhibitors, foam inhibitors, disintegration aids, abrasives, UV absorbers, solvents, antistatic agents, pearlescers and skin protection agents.
  • 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. So are usual Temperatures for detergents and cleaning agents in areas of 1O 0 C for manual compositions over 4O 0 C and 6O 0 C to 95 ° for machine agents or industrial applications. Since the temperature is usually infinitely adjustable in modern washing machines and dishwashers, all intermediate stages of the temperature are included.
  • the ingredients of the respective agents are coordinated. Synergies in terms of cleaning performance are preferred. Particularly preferred in this regard are synergies resulting from the interaction of the two proteases in the composition according to the invention with at least one or more further ingredients. In a further preferred embodiment, these synergies are present in a temperature range between 2O 0 C and 6O 0 C, as well as the proteases contained in the inventive compositions are catalytically active in this temperature range.
  • an agent according to the invention in particular a washing or cleaning agent, further comprises
  • grayness inhibitor 0.01 to 5% by weight of grayness inhibitor and / or
  • the agent may further comprise optical brighteners, preferably from 0.01% to 5% by weight.
  • compositions according to the invention presents no difficulties and can be carried out in a known manner, for example by spray-drying or granulation, enzymes and possibly other thermally sensitive ingredients such as, for example, bleaching agents optionally being added separately later.
  • inventive compositions having an increased bulk density in particular in the range from 650 g / l to 950 g / l, a process comprising an extrusion step is preferred.
  • 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 with compressive forces in the range of about 50 to 100 kN, preferably at 60 to 70 kN.
  • a tablet produced in this way has a weight of 10 g to 50 g, in particular 15 g up 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. Round tablets preferably have a diameter of 30 mm to 40 mm.
  • 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 26x36 mm or 24x38 mm.
  • Liquid or pasty compositions according to the invention in the form of customary 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.
  • Embodiments of the present invention thus comprise all such solid, powdered, liquid, gelatinous or paste-like administration forms of the agents, which if appropriate can also consist of several phases and can be present in compressed or uncompressed form.
  • a further embodiment of the invention therefore represents agents which are characterized in that they are present as a one-component system. Such means preferably consist of one phase. Of course, means according to the invention may also consist of several phases.
  • the washing or cleaning agent is therefore characterized in that it is divided into several components.
  • the solid dosage forms according to the invention also include extrudates, granules, tablets or pouches, which may be present both in large packages and in portions.
  • the agent is present as a free-flowing powder, in particular with a bulk density of 300 g / l to 1200 g / l, in particular 500 g / l to 900 g / l or 600 g / l to 850 g / l.
  • the agent in particular the washing or cleaning agent, is in liquid, gel or pasty 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 agent according to the invention in particular washing or cleaning agent, can be packaged in a container, preferably an air-permeable container, from which it is released shortly before use or during the washing process.
  • a container preferably an air-permeable container, from which it is released shortly before use or during the washing process.
  • at least one, preferably both, of the proteases contained in the agent and / or further ingredients of the agent may be coated with a substance impermeable to the enzyme (s) at room temperature or in the absence of water, which under application conditions of the agent is permeable to the enzyme / the enzymes becomes.
  • Such an embodiment of the invention is therefore characterized in that at least one protease is coated with a substance that is impermeable to the protease at room temperature or in the absence of water.
  • an agent according to the invention is characterized in that it contains the proteases in an amount of from 0.1 ⁇ g to 100 mg, preferably from 1 ⁇ g to 50 mg, more preferably from 10 ⁇ g to 50 mg, particularly preferably from 25 ⁇ g to 25 mg and most preferably from 50 ⁇ g to 20 mg per g of the agent.
  • the agent contains the proteases in an amount of 0.00001 weight percent to 10 weight percent, preferably 0.0001 weight percent to 5 weight percent, more preferably 0.001 weight percent to 5 weight percent, most preferably 0.00025 weight percent to 2.5 Percent by weight and most preferably from 0.005% to 2% by weight.
  • the quantities shown contain the first protease and the second protease.
  • compositions according to the invention may contain exclusively the two proteases described above. Alternatively, they may also contain other proteases or other enzymes in a concentration effective for the effectiveness of the agent.
  • a further subject of the invention thus represents agents which further comprise one or more further enzymes, wherein in principle all enzymes established in the prior art for these purposes can be used.
  • enzymes preferably used are all enzymes that can develop a catalytic activity in the agent of the invention, in particular proteases, amylases, cellulases, hemicellulases, mannanases, tannases, xylanases, xanthanases, ß-glucosidases, carrageenases, oxidases, oxidoreductases, pectin-degrading Enzymes (pectinases) or lipases, and preferably mixtures thereof.
  • proteases amylases, cellulases, hemicellulases, mannanases, tannases, xylanases, xanthanases, ß-glucosidases, carrageenases, oxidases, oxidoreductases, pectin-degrading Enzymes (pectinases) or lipases, and preferably mixtures thereof.
  • pectinases pec
  • compositions according to the invention preferably contain enzymes in total amounts of 1 ⁇ 10 -8 to 5 percent by weight, based on active protein.
  • the enzymes are from 0.00001 to 5 Wt .-%, more preferably from 0.0001 to 2.5 wt .-%, even more preferably from 0.0001 to 1 wt .-% and particularly preferably from 0.0001 to 0.072 wt .-% in inventive compositions , Wherein each enzyme contained can be present in the stated proportions.
  • the protein concentration can be determined by known methods, for example, the BCA method (bicinchoninic acid, 2,2'-biquinolyl-4,4'-dicarboxylic acid) or the biuret method (AG Gornall, CS Bardawill and MM David, J. Biol. Chem., 177 (1948), pp. 751-766).
  • the further enzymes particularly preferably support the effect of the agent, for example the cleaning performance of a washing or cleaning agent, with regard to certain stains or stains. Most preferably, the enzymes exhibit synergistic effects on their action against certain soils or stains, i. the enzymes contained in the middle composition mutually support each other in their cleaning performance.
  • the agent according to the invention is therefore characterized in that it contains at least one further enzyme which comprises a protease, amylase, cellulase, hemicellulase, mannanase, tannase, xylanase, xanthanase, ⁇ -glucosidase, carrageenase, oxidase, oxidoreductase , Pectin degrading enzyme or lipase.
  • at least one further enzyme which comprises a protease, amylase, cellulase, hemicellulase, mannanase, tannase, xylanase, xanthanase, ⁇ -glucosidase, carrageenase, oxidase, oxidoreductase , Pectin degrading enzyme or lipase.
  • protease activity in such agents can be determined by the method described in Tenside, Vol. 7 (1970), pp. 125-132. It is given in PE (protease units) accordingly.
  • the enzymes used in the compositions according to the invention are either originally derived from microorganisms, for example the genera Bacillus, Streptomyces, Humicola or Pseudomonas, and / or are prepared by suitable biotechnological processes Microorganisms produced, for example, by transgenic expression hosts of the genera Bacillus or by filamentous fungi.
  • a separate subject of the invention is the use of an above-described agent according to the invention for the removal of protease-sensitive stains on textiles or hard surfaces, i. for cleaning textiles or hard surfaces.
  • agents according to the invention can be used, in particular in accordance with the properties described above, to remove proteinaceous impurities from textiles or from hard surfaces.
  • Embodiments include, for example, hand washing, manual removal of stains from fabrics or hard surfaces, or use in conjunction with a machine process.
  • the relevant agents according to the invention preferably detergents or cleaning agents, are provided according to one of the above-described embodiments.
  • a further subject of the invention are processes for the cleaning of textiles or of hard surfaces, in which an agent according to the invention is used at least in one of the process steps.
  • the process for the cleaning of textiles or hard surfaces is accordingly characterized in that an agent according to the invention is used in at least one process step.
  • Methods for cleaning textiles are generally distinguished by the fact that various cleaning-active substances are applied to the items to be cleaned in a plurality of process steps and washed off after the action time, or that the items to be cleaned are otherwise treated with a detergent or a solution of this agent.
  • a single substep of such a process may be for cleaning, in particular for machine cleaning, textiles or hard surfaces therein
  • an enzyme mixture according to the invention is applied as sole cleaning-active component. This represents a particularly simplified embodiment of the present invention.
  • Another object of the invention are methods for the purification of textiles or hard surfaces, which are characterized in that in at least one method step, a first protease and a second protease are proteolytically active, wherein the first protease is selected from a) a protease, the an amino acid sequence which is at least 80% identical to the amino acid sequence given in SEQ ID NO.1; b) a protease comprising an amino acid sequence which is at least 80% identical to the amino acid sequence given in SEQ ID NO. 2; c) a protease which comprises an amino acid sequence which is at least 80% identical to the amino acid sequence given in SEQ ID NO. 2 and additionally has at least one additional amino acid in comparison with SEQ ID NO. 2; and the second protease comprises an amino acid sequence which is at least 80% identical to the amino acid sequence given in SEQ ID NO.3.
  • the first protease is selected from a) a protease, the an amino acid sequence which is at least 80%
  • the first protease is selected from a) a protease which comprises an amino acid sequence which is increasingly preferably at least 82.5%, 85%, 87.5% to the amino acid sequence given in SEQ ID NO. 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5%, and most preferably 100% identical.
  • a protease comprising an amino acid sequence which is more preferably at least 82.5%, 85%, 87.5%, 90%, 91%, 92%, 93%, of the amino acid sequence given in SEQ ID NO. 94%, 95%, 96%, 97%, 98%, 99%, 99.5% and most preferably 100% identical.
  • a protease comprising an amino acid sequence which is more preferably at least 82.5%, 85%, 87.5%, 90%, 91%, 92%, 93% to the amino acid sequence given in SEQ ID NO. Is 94%, 95%, 96%, 97%, 98%, 99%, 99.5% and most preferably 100% identical and has at least one additional amino acid compared to SEQ ID NO.2.
  • the method is characterized in that the additional amino acid of the first protease is inserted in one of the following positions in the counting manner according to SEQ ID NO.
  • proteases which are used according to the invention as the first protease in a corresponding method, have a corresponding insertion of at least one amino acid or of a plurality of amino acids, preferably at one or more of said positions on.
  • agents according to the invention - for example for proteases, which are described in the European patent application EP 1 032 655.
  • These have, for example, at least one additional amino acid in the counting method according to SEQ ID NO.
  • proteases which have an insertion of at least one amino acid and can be used according to the invention as the first protease, are derived from the international patent application WO 00/37599 and the international patent application WO 01/44452.
  • the method is characterized in that the second protease comprises an amino acid sequence which is increasingly preferably at least 82.5% to the amino acid sequence given in SEQ ID NO. 85%, 87.5%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5%, and most preferably 100% identical is.
  • the proteases are preferably used in an amount of from 40 ⁇ g to 4 g, preferably from 50 ⁇ g to 3 g, particularly preferably from 100 ⁇ g to 2 g and very particularly preferably from 200 ⁇ g to 1 g per application.
  • the quantities shown contain the first protease and the second protease.
  • methods for textile raw materials, fibers or textiles with natural components are preferred, and especially for those with wool or silk.
  • These may be, for example, processes in which materials for processing in textiles are prepared, for example for anti-fungal finishing, or, for example, for processes which enrich the cleaning of worn textiles with a nourishing component.
  • they are processes for the treatment of textile raw materials, fibers or textiles with natural constituents, in particular with wool or silk.
  • a further inventive subject matter is the use of a first protease and a second protease for the purification of textiles or hard surfaces, characterized in that the first protease is selected from a) a protease comprising an amino acid sequence corresponding to the amino acid sequence shown in SEQ ID NO. 1 amino acid sequence is at least 80% identical; b) a protease comprising an amino acid sequence which is at least 80% identical to the amino acid sequence given in SEQ ID NO. 2; c) a protease which comprises an amino acid sequence which is at least 80% identical to the amino acid sequence given in SEQ ID NO. 2 and additionally has at least one additional amino acid in comparison with SEQ ID NO. 2; and the second protease comprises an amino acid sequence which is at least 80% identical to the amino acid sequence given in SEQ ID NO.3.
  • combinations of proteases according to the invention can be used, in particular because of the properties described above, to eliminate proteinaceous impurities from textiles or from hard surfaces.
  • Embodiments include, for example, hand washing, manual removal of stains from fabrics or hard surfaces, or use in conjunction with a machine process.
  • the subject combinations of proteases of this invention are provided according to any of the above recipes.
  • the first protease is selected from a) a protease which comprises an amino acid sequence which is more preferably at least 82.5%, 85%, 87.5% to the amino acid sequence given in SEQ ID NO. 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5%, and most preferably 100% identical.
  • a protease comprising an amino acid sequence which is more preferably at least 82.5%, 85%, 87.5%, 90%, 91%, 92%, 93% to the amino acid sequence given in SEQ ID NO. Is 94%, 95%, 96%, 97%, 98%, 99%, 99.5% and most preferably 100% identical and has at least one additional amino acid compared to SEQ ID NO.2.
  • the use is characterized in that the additional amino acid of the first protease is inserted in one of the following positions in the counting manner according to SEQ ID NO.2: 33, 34, 35, 36, 37, 38, 39, 40 , 41, 42, 43, 95, 96, 97, 98, 99, 100, 101, 102, 103, 125, 126, 127, 126, 129, 130, 131, 132, 153, 154, 155, 156, 157 , 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 181, 182, 183, 184, 185, 186, 187, 188, 189 , 190, 191, 192, 193, 194, 195, 202, 203, 204, 218, 219.
  • proteases which are useful according to the invention as the first protease, have a corresponding insertion of at least one amino acid.
  • examples of such enzymes which have a corresponding insertion are already discussed above in the present application for the agents according to the invention and the methods according to the invention.
  • the use is characterized in that the second protease comprises an amino acid sequence which is increasingly preferably at least 82.5% to the amino acid sequence given in SEQ ID NO. , 85%, 87.5%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% and most preferably 100% is identical.
  • a preferred use of a combination of proteases according to the invention is characterized in that the proteases are present in an amount of from 40 ⁇ g to 4 g, preferably from 50 ⁇ g to 3 g, more preferably from 100 ⁇ g to 2 g and most preferably from 200 ⁇ g to 1 g per application are used.
  • the quantities shown contain the first protease and the second protease.
  • Example 1 Synergistic Performance of the Protease Variant S99SD + S99A According to WO 01/44452 and the Alkaline Protease from Bacillus Gibsonii (DSM 14391)
  • the pure enzyme granules and mixtures of the protease disclosed as Subtilase 309 (also referred to as Subtilisin 309 or Savinase®) variant S99SD + S99A are disclosed in international patent application WO 01/44452 (see especially Examples 1 and 2 of WO 01/44452) as a representative of the first protease and the alkaline protease from Bacillus gibsonii (DSM 14391) as a representative of the second protease.
  • the mixture was calculated in proportion to the insertions of the single enzymes.
  • the granules of the alkaline protease from Bacillus gibsonii (DSM 14391) were prepared according to WO 97/40128 A1 and coated according to WO 98/26037 A2.
  • Table 1 Results of the performance comparison of a mixture of proteases compared to the pure proteases in a machine dishwashing detergent.
  • the pure enzyme granules and mixtures of the alkaline protease from Bacillus lentus F49 were incorporated as a representative of the first protease and the alkaline protease from Bacillus gibsonii (DSM 14391) as a representative of the second protease.
  • the mixture was calculated in proportion to the insertions of the single enzymes.
  • the granules of the alkaline protease from Bacillus gibsonii (DSM 14391) were prepared according to WO 97/40128 A1 and coated according to WO 98/26037 A2.
  • Table 2 Results of the performance comparison of higher concentrated granules with low concentrated granules in a machine dishwashing detergent.
  • Amylase 0 - 0.072 pure protein
  • formulations of solid detergents with this framework formulation were produced within the scope of the invention:
  • Amylase 0 - 0.072 pure protein
  • Pectinase 0 - 0.072 pure protein
  • Linear alkylbenzene sulfonic acid 0-20%
  • Amylase 0 - 0.072 pure protein
  • Pectinase 0 - 0.072 pure protein
  • Amylase 0 - 0.072 pure protein
  • Pectinase 0 - 0.072 (pure protein) Balance water, optical brightener, salts

Abstract

L'invention concerne des compositions d'agents, notamment des agents de lavage et de nettoyage, qui contiennent une première protéase du type = BLAP = (protéase alcaline issue de Bacillus lentus) et une deuxième protéase du type = protéase alcaline issue de Bacillus gibsonii (DSM 14391) =. L'invention concerne en outre desprocédés de nettoyage dans lesquels ces agents sont employés, des utilisations de ces agents et de ces procédés de nettoyage, et des utilisations de ces protéases.
PCT/EP2008/060204 2007-08-10 2008-08-04 Agents contenant des protéases WO2009021867A2 (fr)

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DE200710038031 DE102007038031A1 (de) 2007-08-10 2007-08-10 Mittel enthaltend Proteasen

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