Recombinant polyester-containing agent

Abstract

The present invention relates to the field of enzyme technology, particularly to the anti-pilling effect of enzymes used, for example, in detergents or cleaning agents. The present invention relates to an agent containing a polyesterase as defined herein, particularly a detergent or cleaning agent. The present invention further relates to a method for cleaning textiles and the use of the agent according to the present invention for removing stains. The present invention also relates to the use of a polyesterase as described herein in an agent, preferably a detergent or cleaning agent, for reducing the pilling effect and preventing graying.

Description

Technology Field

[0001] The present invention relates to the field of enzyme technology, particularly to the anti-pilling effect of enzymes used, such as in detergents or cleaning agents. The present invention relates to an agent containing a polyesterase as defined herein, particularly a detergent or cleaning agent. The present invention further relates to a method for cleaning textiles and the use of the agent according to the present invention for removing stains. Furthermore, the present invention relates to the use of a polyesterase as described herein in an agent, preferably a detergent or cleaning agent, for reducing the pilling effect and preventing graying. Background Technology

[0002] All types of textiles will develop pilling over time after being washed multiple times. Pilling refers to the formation of nodules or lints within the fabric. These small pieces of lint are common, particularly in staple fiber fabrics. However, pilling occurs less frequently in long fibers and twisted fibers. Generally, these nodules are caused by loose fibers within the fabric or fibers that have loosely escaped from the fabric. Synthetic fibers tend to pill more than natural fibers because they have smoother surfaces, allowing them to escape from the fabric more quickly than coarser natural fibers. In the case of wool fabrics, these fibers form nodules on the surface by "tangling" primarily due to mechanical friction.

[0003] The primary impact of pilling is a negative visual effect. Due to the formation of nodules on the surface, the fabric appears worn at first glance and looks older than it actually is. Additionally, colored textiles appear less vibrant. In contrast, the functionality of the fabric is hardly or completely impaired. Pilling occurs particularly in areas subjected to high mechanical stress, typically around the shoulders and waist. In these stress-affected areas, the fabric continues to thin, posing a risk of holes or even tearing. Due to undesirable pilling, correspondingly damaged textiles are rejected and discarded by consumers much faster than necessary relative to their functionality.

[0004] Moreover, textiles tend to turn gray when washed. This is because both dirt and detached pigments are released from colored clothes during the washing process. Although attempts have been made to retain these dirt and pigments in the washing solution using various detergent components, it is often impossible to prevent the dirt / pigment from adhering to the clothes and remaining thereon. This is known as the so-called graying effect. This is particularly pronounced in some synthetic fibers, such as polyamides as well as polyesters.

[0005] Technical solutions to reduce pilling effects have so far only been available for cotton textiles. Cellulase is used in detergents to reduce pilling effects (DE 69632910 T3). This means that since cellulase is used in detergents to exhibit anti-pilling or anti-graying effects, it ensures that clothes look new for a longer period of time. However, cellulase functions only on cotton textiles. For other textiles, such as polyester textiles, there is no comparable method to reduce pilling. Therefore, in order to keep clothes looking new for as long as possible—that is, to keep colors vibrant, preserve shape, and maintain a smooth and undamaged surface—a solution to reduce pilling in textiles, especially those containing synthetic fibers such as polyester, is desirable and required.

[0006] Surprisingly, the inventors of the present invention have discovered that fusing a polyesterase with a specific peptide anchor molecule having a binding affinity for polyesters can significantly enhance the performance of the polyesterase. While this effect has been demonstrated using the exemplary polyesterases described herein, it may also be applicable to other enzymes. Thermomonospora curvata, used and described herein ( Thermomonospora curvata ) from DSM 43183 Polyesterase is active under washing process conditions and possesses various nutrient-providing properties for PET textiles. The polyesterase used exhibits rapid PET degradation at 40°C. The enzyme prevents pilling in new polyester textiles or, when combined with cellulase, promotes this effect in polyester / cotton blend textiles. Additionally, already formed pilling can be reduced, meaning that a "restoration" effect can occur. Polyesterase also prevents graying of white laundry and fading / graying of colored laundry. Furthermore, all these positive washing properties are achieved without significantly damaging the fibers when used in appropriate amounts. The present invention relates to the fact that the concentration used can be significantly reduced without loss of performance by enhancing the performance of said enzyme through fusion of this type of enzyme with a peptide having an affinity for polyester fibers. Specific details for implementing the invention

[0007] Therefore, in a first aspect, the present invention relates to an agent, particularly a detergent or cleaning agent, characterized by containing a polyesterase covalently bonded to at least one heterogeneous peptide sequence having an affinity for polyester. The peptide sequence having an affinity for polyester is not naturally linked to the polyesterase, that is, it is heterogeneous to the polyesterase sequence. Therefore, the enzyme described herein is not a naturally occurring enzyme, but is preferably a fusion protein comprising a polyesterase and a peptide sequence having an affinity for polyester.

[0008] In a further aspect, the present invention relates to a method for cleaning a textile, characterized by using an agent according to the present invention in at least one method step. The textile is preferably a polyester-containing textile or is made of polyester.

[0009] In another aspect, the present invention further relates to the use of an agent as described herein for removing stains, preferably a detergent or cleaning agent, particularly preferably a liquid detergent.

[0010] Additionally, further aspects of the present invention include the use of the polyester agent described herein for reducing the pilling effect and / or enhancing the anti-graying effect of an agent containing the polyester agent, preferably a detergent or cleaning agent, particularly preferably a liquid detergent.

[0011] As used herein, “at least one” means one or more, e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or more. With respect to a component or compound, unless otherwise noted, such expression does not mean the absolute number of molecules, but rather the number of molecules of different types corresponding to the relevant definition of the component or compound. Thus, “at least one polyesterase” means that at least one type of polyesterase is contained, not that at least one polyesterase molecule is contained.

[0012] With respect to peptide sequences, the term "heterogeneous" as used herein means that a peptide sequence having an affinity for polyester does not naturally occur in combination with polyesterase. Therefore, the enzyme described herein is a non-natural hybrid of polyesterase and a peptide not combined with polyesterase.

[0013] As used herein, "affinity for polyester" means that the corresponding peptide sequence binds to polyester, in particular polyester fibers, such as polyester fibers used in textiles, under suitable conditions, i.e., typically non-denaturing conditions, where the binding affinity is greater than the affinity of a reference sequence that does not have a specific affinity for polyester. In various embodiments, the dissociation constant K dThis affinity, expressed as, is at least 10 -3 mol / L, preferably at least 10 -4 , at least 10 -5 , at least 10 -6 or at least 10 -7 It can be mol / L.

[0014] When various combined or individual amino acid sequences are referred to herein, unless otherwise noted, such sequences are always expressed in an orientation from the N-terminus to the C-terminus. Furthermore, unless otherwise noted, individual amino acids or amino acid sequences are joined to each other by peptide bonds. Thus, in the expression SEQ ID NO:1-SEQ ID NO:6-SEQ ID NO:2, the hyphen indicates, for example, that these three sequences are fused to each other by peptide bonds.

[0015] In various embodiments of the present invention, a peptide sequence having an affinity for polyester is directly covalently linked to a polyesterase, that is, the first and / or last amino acid of the polyesterase is linked to the terminal amino acid of the peptide sequence by a peptide bond. Alternatively, the linkage may also be achieved by a linker, particularly a peptide linker. Such linkage by a peptide linker is preferred. Suitable linkers are known in the prior art and may be static / rigid or flexible. These characteristics are determined by the secondary structure of the linker; for example, a rigid linker may have an alpha helix as a secondary structure. In various embodiments, the peptide linker sequence is flexible and has no secondary structure or has only short secondary structure elements.

[0016] Typically, this type of peptide linker has a length of 1 to 100 amino acids, preferably 2 to 30 amino acids, more preferably 5 to 25 amino acids. In various embodiments of the present invention, the linker sequence is AEAAAKEAAAKEAAAKA (SEQ ID NO:6) or PPGGNRGTTTTRRPATTTGSSPGP (SEQ ID NO:7), and at least 70%, preferably 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 90.5%, 91%, 91.5%, 92%, 92.5%, 93%, 93.5%, 94%, 94.5%, 95%, 95.5%, 96%, 96.5%, 97%, with respect to the described reference sequence. It is selected from its congeners having sequence identity of 97.5%, 98%, 98.5%, 98.8%, 99.0%, 99.2%, 99.4%, or 99.6%.

[0017] In various embodiments of the present invention, such linker sequences may also contain sequences recognized by proteases as specific interfaces. This type of protease interface makes it possible to separate the individual components of polyesterase from one another, that is, to separate them into peptide sequences having an affinity for the enzyme and polyester.

[0018] A peptide sequence having an affinity for polyester may be located at the N- and / or C-terminus relative to the polyester sequence. It is preferable that the peptide sequence be linked to the polyester sequence at the C- or N-terminus; the peptide sequence is particularly preferably linked to the C-terminus of the polyester, particularly by a peptide linker.

[0019] In various embodiments of the present invention, a peptide sequence having an affinity for polyester has a length of 5 to 200 amino acids, preferably 10 to 100 amino acids, more preferably 15 to 60 amino acids. The sequence may include sequence elements that are repeated multiple times.

[0020] In various embodiments, the peptide sequence having an affinity for polyester is an antimicrobial or antifungal peptide, in particular those having a β-sheet structure, e.g., tacitatin, androctonin, tanatin, and bacterial antimicrobial peptide, e.g., Bacillus subtilis ( B. subtilis It is selected from LCIs from ). Examples of suitable peptide sequences include at least one of the sequences given in SEQ ID NO:2 to SEQ ID NO:5 or their functional homologues, or are composed thereof.

[0021] "Functional homologues" used in this context are at least 70%, preferably 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 90.5%, 91%, 91.5%, 92%, 92.5%, 93%, 93.5%, 94%, 94.5%, 95%, 95.5%, 96%, 96.5%, 97%, 97.5%, 98%, 98.5%, 98.8%, 99.0%, 99.2%, with respect to the described reference sequence. It relates to a sequence having 99.4% or 99.6% the same function and having an affinity for polyester, that is, at least 50%, preferably at least 75%, of the affinity of the reference sequence.

[0022] In various embodiments, the polyesterase is linked at the C-terminus to one of the peptide linkers described above, in particular SEQ ID NO:7 or its homologue, and the peptide linker is also linked at the C-terminus to a peptide sequence having an affinity for polyester, in particular one of the sequences of SEQ ID NO:2 to SEQ ID NO:5 or its functional homologue.

[0023] The following combination, expressed as an orientation from the N-terminus to the C-terminus, is a desirable combination:

[0024] Polyesterase SEQ ID NO:7-SEQ ID NO:2

[0025] Polyesterase SEQ ID NO:7-SEQ ID NO:3

[0026] Polyesterase SEQ ID NO:7-SEQ ID NO:4

[0027] Polyesterase SEQ ID NO:7-SEQ ID NO:5

[0028] Polyesterase SEQ ID NO:6-SEQ ID NO:2

[0029] Polyesterase SEQ ID NO:6-SEQ ID NO:3

[0030] Polyesterase SEQ ID NO:6-SEQ ID NO:4

[0031] Polyesterase SEQ ID NO:6-SEQ ID NO:5

[0032] In various preferred embodiments of the present invention, the polyesterase is a polyesterase having at least 70% sequence identity with the amino acid sequence given in SEQ ID NO:1, 11, or 12 over its entire length. In further preferred embodiments, the polyesterase contained in the agent according to the present invention comprises, or is substantially composed of, or is composed of the amino acid sequence given in SEQ ID NO:1, 11, or 12. In various embodiments of the present invention, the present invention also comprises a polyesterase derived from the amino acid sequence according to SEQ ID NO:1, 11, or 12, for example by mutagenesis.

[0033] In various embodiments of the present invention, the polyesterase comprises, over its entire length, the amino acid sequence given in SEQ ID NO:1, 11 or 12 and at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 90.5%, 91%, 91.5%, 92%, 92.5%, 93%, 93.5%, 94%, 94.5%, 95%, 95.5%, 96%, 96.5%, 97%, 97.5%, 98%, 98.5%, It contains or consists of 98.8%, 99.0%, 99.2%, 99.4%, or 99.6% identical amino acid sequences.

[0034] In various embodiments, the present invention relates to the following enzyme (oriented from the N-terminus to the C-terminus):

[0035] SEQ ID NO:1-SEQ ID NO:7-SEQ ID NO:2 (SEQ ID NO:9)

[0036] SEQ ID NO:1-SEQ ID NO:7-SEQ ID NO:3

[0037] SEQ ID NO:1-SEQ ID NO:7-SEQ ID NO:4

[0038] SEQ ID NO:1-SEQ ID NO:7-SEQ ID NO:5

[0039] SEQ ID NO:1-SEQ ID NO:6-SEQ ID NO:2 (SEQ ID NO:8)

[0040] SEQ ID NO:1-SEQ ID NO:6-SEQ ID NO:3

[0041] SEQ ID NO:1-SEQ ID NO:6-SEQ ID NO:4

[0042] SEQ ID NO:1-SEQ ID NO:6-SEQ ID NO:5

[0043] SEQ ID NO:11-SEQ ID NO:7-SEQ ID NO:2

[0044] SEQ ID NO:11-SEQ ID NO:7-SEQ ID NO:3

[0045] SEQ ID NO:11-SEQ ID NO:7-SEQ ID NO:4

[0046] SEQ ID NO:11-SEQ ID NO:7-SEQ ID NO:5

[0047] SEQ ID NO:11-SEQ ID NO:6-SEQ ID NO:2

[0048] SEQ ID NO:11-SEQ ID NO:6-SEQ ID NO:3

[0049] SEQ ID NO:11-SEQ ID NO:6-SEQ ID NO:4

[0050] SEQ ID NO:11-SEQ ID NO:6-SEQ ID NO:5

[0051] SEQ ID NO:12-SEQ ID NO:7-SEQ ID NO:2

[0052] SEQ ID NO:12-SEQ ID NO:7-SEQ ID NO:3

[0053] SEQ ID NO:12-SEQ ID NO:7-SEQ ID NO:4

[0054] SEQ ID NO:12-SEQ ID NO:7-SEQ ID NO:5

[0055] SEQ ID NO:12-SEQ ID NO:6-SEQ ID NO:2

[0056] SEQ ID NO:12-SEQ ID NO:6-SEQ ID NO:3

[0057] SEQ ID NO:12-SEQ ID NO:6-SEQ ID NO:4

[0058] SEQ ID NO:12-SEQ ID NO:6-SEQ ID NO:5

[0059] In various other embodiments,

[0060] (a) The polyesterase can be obtained by single or multiple conservative amino acid substitution from the polyesterase defined above as a starting molecule and / or;

[0061] (b) The polyesterase can be obtained by fragmentation or deletion, insertion or substitution mutagenesis from the polyesterase defined above as a starting molecule and comprises an amino acid sequence that matches the starting molecule over a length of at least 210, 220, 230, 240, 245, 250, 255, 256, 257, 258, 259, 260 or 261 consecutive amino acids.

[0062] The agent according to the present invention contains polyesterase in each case, preferably in an amount of 0.00001 to 1 wt.% based on the active protein, more preferably in an amount of 0.0001 to 0.5 wt.%, and particularly preferably in an amount of 0.001 to 0.1 wt.%.

[0063] The identity of nucleic acid or amino acid sequences is determined by sequence comparison. Such sequence comparison is based on the BLAST algorithm, which is established in the prior art and is commonly used (see, for example, [Altschul et al. (1990): "Basic local alignment search tool," J. Mol. Biol. 215:403-410] and [Altschul et al. (1997): "Gapped BLAST and PSI-BLAST: a new generation of protein database search programs;" Nucleic Acids Res. 25:3389-3402]), and in principle occurs by associating similar sequences of nucleotides or amino acids within the nucleic acid or amino acid sequence. The assignment of related positions, represented in a table, is referred to as alignment. Another algorithm available in the prior art is the FASTA algorithm. Sequence comparison (alignment), particularly multiple sequence comparison, is performed using computer programs. For example, the Clustal series (see, e.g., Chenna et al. (2003) "Multiple sequence alignment with the Clustal series of programs," Nucleic Acid Res. 31:3497-3500), T-Coffee (see, e.g., Notredame et al. (2000) "T-Coffee: A novel method for multiple sequence alignments," J. Mol. Biol. 302:205-217), or programs based on these programs or algorithms are frequently used. A computer program Vector NTI® Suite 10, equipped with an AlignX module based on ClustalW for sequence comparison and with predetermined basic parameters.Sequence comparison (alignment) using 3 (Invitrogen Corporation, 1600 Faraday Avenue, Carlsbad, California, USA) is also possible.

[0064] Such comparisons also allow for drawing conclusions about the similarity of the compared sequences. This is typically given as percent identity, that is, the ratio of identical nucleotide or amino acid residues in the sequence or in alignment at corresponding positions. A broader concept of homology considers conserved amino acid exchanges in the case of amino acid sequences—namely, amino acids with similar chemical activity—because they typically exert similar chemical activity within proteins. Therefore, the similarity between compared sequences can also be expressed as percent homology or percent similarity. Information on identity and / or homology can be provided for the entire polypeptide or gene, or for just individual regions. Thus, homologous or identity regions of different nucleic acid or amino acid sequences are defined by matching in the sequence. These regions often have the same function. They can be small and contain only a few nucleotides or amino acids. Often, these small regions perform functions essential to the overall activity of the protein. Therefore, it may be convenient to relate sequence matching to only arbitrarily small individual regions. However, unless otherwise noted, identity or homology information in this application relates to the total length of the specific nucleic acid or amino acid sequence described.

[0065] In various embodiments, the polyesterase comprises an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 90.5%, 91%, 91.5%, 92%, 92.5%, 93%, 93.5%, 94%, 94.5%, 95%, 95.5%, 96%, 96.5%, 97%, 97.5%, 98%, 98.5%, 98.8%, 99.0%, 99.2%, 99.4%, or 99.6% homology with the amino acid sequence given in SEQ ID NO:1, 11, or 12 over its entire length.

[0066] In a further embodiment of the present invention, the polyesterase is characterized in that its anti-pilling performance is not significantly reduced compared to that of a polyesterase comprising an amino acid sequence corresponding to the amino acid sequence given in SEQ ID NO:1, 11 or 12, i.e., at least 70%, 75%, 80%, 85%, 90%, or 95% of the reference anti-pilling performance. The anti-pilling performance may be determined in a washing system containing a detergent and polyesterase in an amount of 4.5 to 7.0 grams per liter of washing liquid, wherein the polyesterase to be compared is used at the same concentration (based on active protein) and the anti-pilling performance is determined as described herein. For example, the washing process may be performed for 60 minutes at a temperature of 60° and the water may have a water hardness (German hardness) of 15.5° to 16.5°. The concentration of polyesterase in the detergent intended for this laundry system is 0.00001 to 1 wt.%, preferably 0.0001 to 0.5 wt.%, and particularly preferably 0.001 to 0.1 wt.% based on the purified active protein.

[0067] A preferred liquid detergent for such a laundry system is composed as follows (all figures are expressed in wt.%): 4.4% alkylbenzenesulfonic acid, 5.6% anionic surfactant, 2.4% C 12-C 18 Na salt of fatty acid, 4.4% non-ionic surfactant, 0.2% phosphonate, 1.4% citric acid, 0.95% NaOH, 0.01% antifoaming agent, 2% glycerol, 0.08% preservative, 1% ethanol, 1.6% enzyme mixture (protease, amylase, cellulase, mannanse), and the remainder being deionized water. Preferably, the amount of liquid detergent used is 4.5 to 6.0 grams per liter of laundry liquid, for example, 4.7, 4.9, or 5.9 grams per liter of laundry liquid. Washing is preferably performed in a pH range of pH 8 to pH 10.5, preferably pH 8 to pH 9.

[0068] In the context of the present invention, the anti-pilling performance is determined at 60°C using a liquid detergent as described above, and the washing process is preferably performed for 60 minutes.

[0069] Pilling prevention performance can be tracked using visual sampling. In this case, a group of testers assigns a grade of 1 to 5 to the laundry to be inspected. A value of 1 indicates very severely pilled laundry, and a value of 5 is associated with non-pilling laundry.

[0070] The use of activity-equivalents for related polyesterases ensures that their respective enzymatic properties, such as anti-peeling performance, are considered similar even if the ratio of active substance to total protein (inactivity value) varies. Generally, lower inactivity can be compensated for by adding a larger amount of protein.

[0071] Proteins may be included in a group of immunologically related proteins through reaction with antiserum or specific antibodies. Members of this group are characterized by the fact that they possess the same epitope that is recognized by antibodies. Therefore, they are structurally very similar and are recognized by antiserum or specific antibodies. Accordingly, the present invention further relates to a polyesterase characterized by having at least one, more preferably two, three, or four epitope that matches the polyesterase used in the agent according to the present invention. Due to their immunological similarity, it can be assumed that such a polyesterase is structurally very similar to the polyesterase used in the agent according to the present invention and has a similar function.

[0072] Additional polyesterases used in the agent according to the present invention may undergo additional amino acid changes, particularly amino acid substitutions, insertions, or deletions, compared to the polyesterases described in SEQ ID NO:1, 11, or 12. For example, such polyesterases are developed by targeted genetic modification, i.e., mutagenic methods, and are optimized for specific applications or in relation to specific characteristics (e.g., in relation to their catalytic activity, stability, etc.). Furthermore, the nucleic acid encoding the polyesterase used may be introduced into a recombinant approach and thus can be used to generate a completely new type of polyesterase or other polypeptide.

[0073] The objective is to introduce targeted mutations, such as substitutions, insertions, or deletions, into known molecules to improve the cleaning performance of the enzyme according to the present invention, for example. For this purpose, the surface charge and / or isoelectric point of the molecule and, therefore, the interaction between it and the substrate may be altered. For example, the net charge of the enzyme may be altered to affect substrate binding, particularly when used in laundry and detergents. Alternatively or additionally, the cleaning performance of the polyesterase may be improved by further enhancing its stability through one or more corresponding mutations. The advantageous properties of individual mutations, for example, individual substitutions, may complement each other. Therefore, polyesterases already optimized with respect to specific properties, for example, activity and / or anti-pilling performance, may also be developed within the scope of the present invention.

[0074] Therefore, another object of the present invention is an agent containing a polyesterase characterized by being obtainable by single or multiple conservative amino acid substitutions from the polyesterase described above as a starting molecule. The term "conservative amino acid substitution" means exchanging (substituting) one amino acid residue with another amino acid residue without causing a change in polarity or charge at the position of the exchanged amino acid, for example, exchanging a nonpolar amino acid residue with another nonpolar amino acid residue. Within the scope of the present invention, conservative amino acid substitutions include, for example, the following: G=A=S, I=V=L=M, D=E, N=Q, K=R, Y=F, S=T, G=A=I=V=L=M=Y=F=W=P=S=T. The homology of the polyesterase modified in this way to the polyesterase having SEQ ID NO:1, 11, or 12 is preferably as defined above.

[0075] Alternatively or additionally, the polyesterase is obtainable by fragmentation, deletion, insertion, or substitution mutagenesis from the polyesterase contained in the agent according to the present invention as a starting molecule, and is characterized by comprising an amino acid sequence that matches the starting molecule over a length of at least 210, 220, 230, 240, 245, 250, 255, 256, 257, 258, 259, 260, or 261 consecutive amino acids.

[0076] In various embodiments, the polyesterase obtainable in this manner also has at least 70% sequence identity with the sequence according to SEQ ID NO:1, 11 or 12 even after mutagenesis / substitution.

[0077] For example, it is possible to cause the deletion of individual amino acids at the end or loop of the enzyme without the hydrolytic activity being lost or reduced in the process. Furthermore, such fragmentation or deletion, insertion or substitution mutagenesis can also, for example, reduce the allergenicity of the relevant enzyme and thus improve its overall applicability. Advantageously, the enzyme retains its hydrolytic activity after mutagenesis, that is, its hydrolytic activity corresponds to at least that of the starting enzyme, i.e., in a preferred embodiment, the hydrolytic activity is at least 80%, preferably at least 90%, of the activity of the starting enzyme. Other substitutions may also produce advantageous effects. Both single and multiple consecutive amino acids can be exchanged for other amino acids.

[0078] In various embodiments, the polyesterase may have one or more additional amino acids in addition to the N- or C-terminus of the sequence specified in SEQ ID NO:1, 11, or 12. In certain embodiments, this N-terminal peptide may be a naturally occurring signal peptide for the polyesterase or a single methionine residue. In this type of embodiment, the polyesterase has, for example, the amino acid sequence given in SEQ ID NO:10. In the context of a mature polyesterase according to SEQ ID NO:1, 11, or 12, all embodiments disclosed above are also applicable to the polyesterase of the sequence according to SEQ ID NO:10.

[0079] The object of the present invention is an agent characterized by containing a polyester agent as defined herein. The agent is preferably a laundry agent or a cleaning agent.

[0080] Unless otherwise expressly stated, all percentages mentioned in relation to the compositions / agents described herein relate to wt.% based on the relevant mixture / related agent in each case.

[0081] Within the scope of the present invention, unless otherwise noted, fatty acids and / or fatty alcohols and / or their derivatives preferably represent branched or unbranched carboxylic acids and / or alcohols and / or their derivatives having 6 to 22 carbon atoms. In particular, oxo-alcohols or their derivatives that can be obtained, for example, from a Roelen oxo-synthesis reaction may also be used correspondingly.

[0082] Whenever an alkaline earth metal is mentioned below as the counterion of a monovalent anion, this means that the alkaline earth metal exists naturally in only half the amount of substance—enough to balance the charge—of the anion.

[0083] The object of the present invention is an agent characterized by containing a polyester agent as defined herein. The agent is preferably a laundry agent or a cleaning agent.

[0084] The subject matter of the present invention comprises all conceivable types of detergents or cleaning agents, including both concentrated and undiluted agents, for use on a commercial scale, in washing machines, or in manual washing. This includes, for example, detergents for textiles, carpets, or natural fibers, for which the term detergent is used. In the context of the present invention, detergents and cleaning agents also include auxiliary detergents added to the actual detergent when washing textiles manually or mechanically to achieve additional effects. Furthermore, detergents and cleaning agents according to the present invention also include textile pretreatment agents and posttreatment agents, namely agents that come into contact with the laundry item before the actual washing cycle to loosen, for example, stubborn stains, and agents that provide additional desirable properties to the laundry, such as a pleasant feel, anti-wrinkle, or low static electricity, in a stage after the actual textile washing. In particular, softeners are included in the agents mentioned last.

[0085] The detergent or cleaning agent according to the present invention, which may exist in the form of a powdered solid, additionally compressed particulates, a homogeneous solution, a gel, or a suspension, may contain all known components conventional to such agents in addition to the polyester agent described above, and preferably at least one other component is present in the agent. The agent according to the present invention may contain, in particular, surfactants, builders, bleaches, in particular peroxy compounds, or bleaching activators. They may also contain water-miscible organic solvents, additional enzymes, sequestering agents, electrolytes, pH adjusters and / or additional auxiliary agents, such as optical brighteners, graying inhibitors, foaming agents, as well as dyes and fragrances, and combinations thereof.

[0086] In particular, it is advantageous to combine the agent according to the present invention with one or more additional components, because in a preferred embodiment according to the present invention, such an agent has improved cleaning performance due to the resulting synergy. In particular, such synergy can be achieved by combining the agent according to the present invention with a surfactant and / or builder and / or peroxygen compound and / or bleaching activator.

[0087] The advantageous components of the agent according to the present invention are disclosed in the International Patent Application WO 2009 / 121725, starting from the second-to-last paragraph on page 5 and ending after the second paragraph on page 13. Such disclosures are explicitly referenced and are incorporated into this patent application.

[0088] A person skilled in the art will become aware of these and other aspects, features, and advantages of the present invention by reviewing the following detailed description and claims. Any feature of one aspect of the present invention may be used in any other aspect of the present invention. Furthermore, it will be readily understood that the embodiments included herein are intended to describe and illustrate the present invention but are not intended to limit the present invention, and in particular, that the present invention is not limited to these embodiments. Unless otherwise stated, all percentages described are weight percentages based on the total weight of the composition. Numerical ranges described in the form of "x to y" also include the mentioned values. Where multiple preferred numerical ranges are described in this form, it will be readily understood that all ranges created by combining various endpoints are also included.

[0089] In addition to polyester agents, the agent according to the present invention preferably also contains at least one compound selected from the class of surfactants, particularly anionic and non-ionic, as well as cationic, zwitterionic, or amphoteric surfactants.

[0090] Suitable surfactants are, for example, anionic surfactants of chemical formula (I):

[0091] R-SO3 - Y + (I).

[0092] In this chemical formula (I), R represents a linear or branched, unsubstituted alkyl aryl functional group. Y + represents the 1 / n portion of a monovalent cation or n-valent cation, and in this case, Na + or K + Alkali metal ions including are preferred, and Na + It is most desirable. Additional cation Y + is NH4 + , ½ Zn 2+ , ½ Mg 2+ , ½ Ca 2+ , ½ Mn 2+ It can be selected from , and a mixture thereof.

[0093] As used herein, "alkyl aryl" refers to an organic functional group composed of an alkyl functional group and an aromatic functional group. Typical examples of this type of functional group include, but are not limited to, alkylbenzene functional groups, such as benzyl, butylbenzene functional groups, nonylbenzene functional groups, decylbenzene functional groups, undecylbenzene functional groups, dodecylbenzene functional groups, tridecylbenzene functional groups.

[0094] In various embodiments, this type of surfactant is selected from linear or branched alkylbenzene sulfonates of formula A-1:

[0095]

[0096] Here, R' and R" together contain 9 to 19, preferably 11 to 15, and particularly 11 to 13 C atoms. A very particularly preferred representative can be represented by the formula A-1a:

[0097]

[0098] In various embodiments, the compound of formula (I) is preferably a sodium salt of a linear alkylbenzene sulfonate.

[0099] In the agent according to the present invention, at least one compound from the class of anionic surfactants of formula (I) is contained in the detergent or cleaning agent in an amount of 0.001 to 30 wt.%, preferably 0.001 to 10 wt.%, more preferably 2 to 6 wt.%, and even more preferably 3 to 5 wt.% based on the total weight of the cleaning agent in each case.

[0100] In various embodiments, the agent according to the present invention preferably contains at least one anionic surfactant of the following formula:

[0101] R 1 -O-(AO) n -SO3 - X + (II).

[0102] In this chemical formula (II), R 1 represents a linear or branched, substituted or unsubstituted alkyl, aryl, or alkyl-aryl functional group, preferably a linear, unsubstituted alkyl functional group, particularly preferably a fatty alcohol functional group. Preferred functional group R 1 It is selected from decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecyl, octadecyl, nonadecyl, eicosyl functional groups and mixtures thereof, and a representative one having an even number of C atoms is preferred. Particularly preferred functional group R 1 C 12 -C 18Fatty alcohols, e.g., coconut fatty alcohol, tallow fatty alcohol, lauryl, myristyl, cetyl, or stearyl alcohol or C 10 -C 20 It is derived from oxo-alcohols.

[0103] AO represents an ethylene oxide (EO) group or a propylene oxide (PO) group, preferably an ethylene oxide group. The exponent n represents an integer from 1 to 50, preferably from 1 to 20, particularly from 2 to 10. Very particularly preferably, n represents a number of 2, 3, 4, 5, 6, 7, or 8. X + represents the 1 / n portion of a monovalent cation or n-valent cation, and in this case, Na + or K + Alkali metal ions including are preferred, and Na + is most desirable. Additional cation X + is NH4 + , ½ Zn 2+ , ½ Mg 2+ , ½ Ca 2+ , ½ Mn 2+ It can be selected from , and a mixture thereof.

[0104] Accordingly, in summary, in various embodiments, the agent contains at least one anionic surfactant selected from fatty alcohol ether sulfates of formula A-2:

[0105]

[0106] Here, k = 11 to 19 and n = 2, 3, 4, 5, 6, 7, or 8. A particularly preferred representative is Na-C having two EOs. 12-14 It is a fatty alcohol ether sulfate (k = 11-13, n = 2 in chemical formula A-2).

[0107] In various embodiments, the cleaning agent contains at least one anionic surfactant of formula (II) in an amount of 2 to 10 wt.%, preferably 3 to 8 wt.%, based on the total weight of the cleaning agent.

[0108] Other anionic surfactants that can be used are alkyl sulfates of the following chemical formula:

[0109] R 2 -O-SO3 - X + (III).

[0110] In this chemical formula (III), R 2 represents a linear or branched, substituted or unsubstituted alkyl functional group, preferably a linear, unsubstituted alkyl functional group, particularly preferably a fatty alcohol functional group. Preferred functional group R 2 is selected from decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecyl, octadecyl, nonadecyl, eicosyl functional groups and mixtures thereof, and a representative one having an even number of C atoms is preferred. Particularly preferred functional group R 2 is C 12 -C 18 Fatty alcohols, e.g., coconut fatty alcohol, tallow fatty alcohol, lauryl, myristyl, cetyl, or stearyl alcohol or C 10 -C 20 It is derived from oxo-alcohols. X + represents the 1 / n portion of a monovalent cation or n-valent cation, and in this case, Na + or K + Alkali metal ions including are preferred, and Na + is most desirable. Additional cation X + is NH4 + , ½ Zn 2+ , ½ Mg 2+ , ½ Ca 2+ , ½ Mn 2+ It can be selected from , and a mixture thereof.

[0111] In various embodiments, these surfactants are selected from fatty alcohol sulfates of formula A-3:

[0112]

[0113] Here, k = 11 to 19. A very particularly desirable representative is Na-C 12-14 It is a fatty alcohol sulfate (k = 11-13 in chemical formula A-3).

[0114] In various embodiments, the agent may contain at least one other surfactant in addition to the anionic surfactants described above, particularly surfactants of formulas (I) to (III), or alternatively. The alternative or additional surfactants are particularly additional anionic surfactants, non-ionic surfactants and mixtures thereof, as well as cationic, zwitterionic, and amphoteric surfactants.

[0115] In various embodiments, the agent comprises at least one non-ionic surfactant, in particular at least one fatty alcohol alkoxylate.

[0116] Suitable non-ionic surfactants are those of the following chemical formulas:

[0117] R 3 -O-(AO) m -H (IV),

[0118] Here, R 3 represents a linear or branched, substituted or unsubstituted alkyl functional group, AO represents an ethylene oxide (EO) or propylene oxide (PO) group, and m represents an integer from 1 to 50.

[0119] In the aforementioned chemical formula (IV), R 3 represents a linear or branched, substituted or unsubstituted alkyl functional group, preferably a linear, unsubstituted alkyl functional group, particularly preferably a fatty alcohol functional group. Preferred functional group R 2is selected from decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecyl, octadecyl, nonadecyl, eicosyl functional groups and mixtures thereof, and a representative one having an even number of C atoms is preferred. Particularly preferred functional group R 3 C 12 -C 18 Fatty alcohols, e.g., coconut fatty alcohol, tallow fatty alcohol, lauryl, myristyl, cetyl, or stearyl alcohol or C 10 -C 20 It is derived from oxo-alcohols.

[0120] AO represents an ethylene oxide (EO) group or a propylene oxide (PO) group, preferably an ethylene oxide group. The exponent m represents an integer from 1 to 50, preferably from 1 to 20, particularly from 2 to 10. Very particularly preferably, m represents a number of 2, 3, 4, 5, 6, 7, or 8.

[0121] In summary, the fatty alcohol alkoxylates preferably used are compounds of the following chemical formula:

[0122]

[0123] Here, k = 11 to 19 and m = 2, 3, 4, 5, 6, 7, or 8. A very particularly preferred representative is C having 7 EOs. 12-18 It is a fatty alcohol (k = 11-17, m = 7 in chemical formula (V)).

[0124] Additional non-ionic surfactants that may be contained in the agent described within the meaning of the present invention include, but are not limited to, alkyl glycosides, alkoxylated fatty acid alkyl esters, amine oxides, fatty acid alkanolamides, hydroxy mixed ethers, sorbitan fatty acid esters, polyhydroxy fatty acid amides, and alkoxylated alcohols.

[0125] Suitable amphoteric surfactants are, for example, those with the chemical formula (R iii )(Riv )(R v )N + CH2COO - It is the beta of, where R iii Silver represents an alkyl functional group having 8 to 25, preferably 10 to 21, carbon atoms, wherein a heteroatom or a heteroatom group is optionally interposed, and R iv and R v is the same or different alkyl functional group having 1 to 3 carbon atoms, in particular C 10 -C 18 Alkyl dimethyl carboxymethyl betaine and C 11 -C 17 It represents alkyl amidopropyl dimethyl carboxymethyl betaine.

[0126] Suitable cationic surfactants, in particular, have the chemical formula (R vi )(R vii )(R viii )(R ix )N + X - quaternary ammonium compounds of, for example, didecyl dimethyl ammonium chloride, alkyl benzyl didecyl ammonium chloride and mixtures thereof, where R vi to R ix represents four identical or different, specifically two long-chain and two short-chain, alkyl functional groups, and X - represents an anion, particularly a halide ion. Additional suitable cationic surfactants are quaternary surface-active compounds having sulfonium, phosphonium, iodonium, or arsonium groups, which are also known as antimicrobial active components. By using quaternary surface-active compounds that have antimicrobial activity, agents can be designed that have an antimicrobial effect or that can enhance an antimicrobial effect that may already be present due to other components.

[0127] In various embodiments, the total amount of surfactant based on the weight of the agent is 2 to 30 wt.%, preferably 5 to 25 wt.%, more preferably 10 to 20 wt.%, most preferably 14 to 18 wt.%, wherein the (linear) alkylbenzene sulfonate is present in an amount of up to 0.001 to 30 wt.%, preferably 0.001 to 10 wt.%, more preferably 2 to 6 wt.%, and even more preferably 3 to 5 wt.% based on the weight of the agent.

[0128] The detergent or cleaning agent according to the present invention may contain additional enzymes in addition to polyesterase. These may be hydrolytic enzymes or other enzymes at a concentration convenient for the efficacy of the agent. Accordingly, one embodiment of the present invention relates to an agent comprising one or more enzymes. Any enzyme capable of expressing catalytic activity in the agent according to the present invention, in particular protease, amylase, cellulase, hemicellulase, mannanase, tannanase, xylanase, xanthanase, xyloglucanase, β-glucosidase, pectinase, carrageenase, perhydrolase, oxidase or oxidoreductase or lipase, and mixtures thereof may preferably be used as the enzyme. The enzyme is advantageously 1 x 10⁶ based on the active protein in each case. -8 It is contained in the agent in an amount of to 5 wt.%. More preferably, each enzyme is 1 x 10 based on the active protein. -7The enzyme is contained in the agent according to the present invention in an amount of up to 3 wt.%, 0.00001 to 1 wt.%, 0.00005 to 0.5 wt.%, 0.0001 to 0.1 wt.%, and particularly preferably 0.0001 to 0.05 wt.%. Particularly preferably, the enzyme exhibits synergistic cleaning performance against specific stains or dirt, that is, the enzymes contained in the agent composition support each other in their cleaning performance. The synergistic effect may occur not only between different enzymes but also between one or more enzymes and other components of the agent according to the present invention.

[0129] The amylase(s) are preferably α-amylase. The hemicellulase is preferably β-glucanase, pectinase, pullulanase, and / or mannanase. The cellulase is preferably a cellulase mixture or a single-component cellulase, preferably or mainly endoglucanase and / or cellobiohydrolase. The oxidoreductase is preferably an oxidase, in particular choline-oxidase or perhydrolase.

[0130] The protease used is preferably an alkaline serine protease. These act as non-specific endopeptidases, that is, by hydrolyzing any acid amide bonds within peptides or proteins, they remove protein-containing stains from the article to be cleaned. Their optimal pH is typically within the strong alkaline range.

[0131] Protein concentration can be determined using known methods, e.g., the BCA method (bicinconic acid; 2,2'-biquinolyl-4,4'-dicarboxylic acid) or the Biuret method. Active protein concentration is determined by titrating the active center with a suitable irreversible inhibitor (e.g., phenylmethylsulfonylfluoride (PMSF) in the case of proteases) and determining the residual activity (see reference [M. Bender et al. (1966), J. Am. Chem. Soc. 88(24):5890-5913]).

[0132] In the cleaning agent described herein, the enzyme used may further be combined with an incidental substance, for example, derived from fermentation. In the liquid formulation, the enzyme is preferably used as an enzyme liquid formulation.

[0133] Enzymes are generally available not in the form of pure proteins, but in the form of stabilized, storable, and transportable formulations. Such pre-formulated formulations include, for example, solid formulations obtained through granulation, extrusion, or freeze-drying, or, particularly in the case of liquid or gel agents, enzyme solutions that are advantageously maximally concentrated and / or have a low moisture content and / or are supplemented with stabilizers or other auxiliary agents.

[0134] Alternatively, in the case of both solid and liquid input forms, the enzyme may be encapsulated, for example, by spray-drying or extrusion of an enzyme solution, preferably with a natural polymer, or in the form of a capsule, for example, in which the enzyme is enclosed within a cured gel, or as a core-shell type capsule in which an enzyme-containing core is coated with a water-, air-, and / or chemical-impermeable protective layer. Other active components, such as stabilizers, emulsifiers, pigments, bleaches, or dyes, may be additionally added to the overlaid layer. These capsules are added using essentially known methods, for example, via shaking, roll granulation, or fluid bed processes. These granules are advantageously less dusty, for example, due to the addition of a polymeric film-forming agent, and have storage stability due to the coating.

[0135] In addition, it is possible to combine two or more enzymes so that a single granule exhibits multiple enzyme activities.

[0136] In various embodiments, the agent according to the present invention may comprise one or more enzyme stabilizers. Therefore, the agent according to the present invention may further comprise an enzyme stabilizer selected from the group consisting of, for example, sodium formate, sodium sulfate, lower aliphatic alcohols and boric acid, as well as their esters and salts. Of course, two or more of these compounds may also be used in combination. Salts of the mentioned compounds may also be used in the form of hydrates, such as sodium sulfate decahydrate.

[0137] As used herein, the term “lower aliphatic alcohol” includes monoalcohols, diols, and polyalcohols having up to six carbon atoms. In this context, particularly polyols, e.g., glycerol, (mono)ethylene glycol, (mono)propylene glycol, or sorbitol are referred to as belonging to the lower aliphatic alcohol group, but the invention is not limited thereto.

[0138] In addition to at least one enzyme stabilizer selected from the above group, the agent according to the present invention may also contain at least one additional stabilizer. Such stabilizers are known in the prior art.

[0139] Reversible protease inhibitors protect enzymes contained in laundry or detergents from proteolysis by reversibly inhibiting the enzymatic activity of proteases contained in the agent. Benzamidine hydrochloride, boronic acid or its salts or esters, particularly derivatives having aromatic groups, e.g., ortho-, meta-, or para-substituted phenylboronic acid, particularly 4-formylphenylboronic acid, or salts or esters of the aforementioned compounds are frequently used as reversible protease inhibitors. Peptaldehydes, i.e., oligopeptides having a reduced C-terminus, particularly those consisting of 2 to 50 monomers, are also used for this purpose. Peptide reversible protease inhibitors include, in particular, ovomucoids and leupeptins.

[0140] Other enzyme stabilizers are amino alcohols such as mono-, di-, triethanol-, and -propanolamine and mixtures thereof, up to C 12 It is an aliphatic carboxylic acid, such as succinic acid, other dicarboxylic acids, or a salt of the mentioned acid. Terminal-capped fatty acid amide alkoxylates are also suitable for this purpose. Some organic acids used as builders can also stabilize the enzyme. Calcium and / or magnesium salts, such as calcium acetate, are also used for this purpose.

[0141] Polyamide oligomers or polymeric compounds, such as lignin, water-soluble vinyl copolymers or cellulose ethers, acrylic polymers, and / or polyamides, in particular, stabilize enzyme preparations against physical influences or pH fluctuations. Polymers containing polyamine N-oxides act simultaneously as enzyme stabilizers and as antiseptic agents. Other polymeric stabilizers are linear C8-C 18 It is a polyoxyalkylene. Alkyl polyglycosides can also stabilize the enzymatic components of the agent according to the present invention and, preferably, further enhance their performance. The cross-linked N-containing compound preferably performs a dual function as a pollutant release agent and an enzyme stabilizer. The hydrophobic non-ionic polymer particularly stabilizes any cellulase that may be contained.

[0142] Reducing agents and antioxidants enhance the stability of enzymes against oxidative disintegration; for this purpose, sulfur-containing reducing agents, such as sodium sulfite and reducing sugars, are common.

[0143] In one embodiment, the agent according to the present invention is a liquid and contains water as the main solvent, i.e., they are aqueous agents. The water content of the aqueous composition according to the present invention is typically 15 to 70 wt.%, preferably 20 to 60 wt.%. In various embodiments, the water content is greater than 5 wt.%, preferably greater than 15 wt.%, and particularly preferably greater than 50 wt.%, based on the total amount of agent in each case.

[0144] Additionally, a non-aqueous solvent may be added to the agent. Suitable non-aqueous solvents include monohydric or polyhydric alcohols, alkanolamines, or glycol ethers, provided they are miscible with water within the stated concentration ranges. Preferably, the solvent is ethanol, n-propanol, i-propanol, butanol, glycol, propanediol, butanediol, methylpropanediol, glycerol, diglycol, propyl diglycol, butyl diglycol, hexylene glycol, ethylene glycol methyl ether, ethylene glycol ethyl ether, ethylene glycol propyl ether, ethylene glycol mono-n-butyl ether, diethylene glycol methyl ether, diethylene glycol ethyl ether, propylene glycol methyl ether, propylene glycol ethyl ether, propylene glycol propyl ether, dipropylene glycol monomethyl ether, dipropylene glycol monoethyl ether, methoxytriglycol, ethoxytriglycol, butoxytriglycol, 1-butoxyethoxy-2-propanol, 3-methyl-3-methoxybutanol, propylene-glycol-t-butyl ether, It is selected from di-n-octyl ether and mixtures of such solvents.

[0145] One or more non-aqueous solvents are typically contained in an amount of 0.1 to 10 wt.%, preferably 1 to 8 wt.%, based on the total composition.

[0146] In addition to the components mentioned above, the agent according to the present invention may contain other components that further improve the practical and / or aesthetic properties of the detergent. These include, for example, additives to improve flow and drying behavior, additives to control viscosity and / or additives for stabilization, and other adjuvants and additional substances conventional in detergents, such as UV stabilizers, perfumes, pearlescent agents, dyes, corrosion inhibitors, preservatives, brine, organic salts, disinfectants, structured polymers, defoaming agents, encapsulated components (e.g., encapsulated perfumes), pH adjusters, and skin-touch-improving or nourishing additives.

[0147] The agent according to the present invention, in particular a washing agent or a cleaning agent, preferably contains at least one water-soluble and / or water-insoluble, organic and / or inorganic builder.

[0148] Builders that can generally be used include, in particular, aminocarboxylic acids and their salts, zeolites, silicates, carbonates, organic (co)builders, and, provided there is no ecological bias regarding their use, also phosphonates. However, the agent preferably does not contain phosphonates.

[0149] Water-soluble organic builders include polycarboxylic acids, particularly citric acid and saccharic acid; monomeric and polymeric aminopolycarboxylic acids, particularly methylglycine diacetic acid, nitrilotriacetic acid, ethylenediaminetetraacetic acid and polyaspartic acid; polyphosphonic acids, particularly aminotris(methylenephosphonic acid), ethylenediaminetetrakis(methylenephosphonic acid) and 1-hydroxyethane-1,1-diphosphonic acid; polymeric hydroxy compounds, such as dextrin; and polymeric (poly)carboxylic acids, polymeric acrylic acid, methacrylic acid, maleic acid; and mixed polymers thereof which may also contain small amounts of polymerizable materials that do not have carboxylic acid functional groups. Less desirable but suitable compounds of this class are copolymers of acrylic acid or methacrylic acid with vinyl ethers, such as vinyl methyl ether, vinyl ester, ethylene, propylene, and styrene, wherein the proportion of acid is at least 50 wt.%. The organic builder may be used, particularly when preparing liquid agents, in the form of an aqueous solution, preferably in the form of a 30 to 50 wt.% aqueous solution. All of the mentioned acids are generally used in the form of their water-soluble salts, particularly their alkali salts.

[0150] The organic builder may be contained in an amount up to 40 wt.%, particularly up to 25 wt.%, preferably 1 to 8 wt.%, if desired. An amount close to the mentioned upper limit is preferably used in the agent according to the present invention in the form of a paste or liquid, particularly containing water. The laundry post-treatment agent according to the present invention, such as a softener, may optionally also not contain the organic builder.

[0151] Suitable water-soluble inorganic builder materials are, in particular, alkali silicates, and, if there are no concerns regarding their use, also polyphosphates, preferably sodium triphosphates. Where desired, particularly crystalline or amorphous alkali aluminosilicates may be used as water-insoluble water-dispersible inorganic builder materials in amounts up to 50 wt.%, preferably 40 wt.% or less, and in liquid agents in amounts particularly 1 to 5 wt.%. Among these, detergent-quality crystalline sodium aluminosilicates, particularly zeolites A, P, and optionally X, are preferred. Amounts close to the mentioned upper limit are preferably used in solid particulate agents. Suitable aluminosilicates do not particularly contain particles with a particle size greater than 30 μm, and preferably contain at least 80 wt.% of particles with a size of less than 10 μm.

[0152] Suitable substitutes or partial substitutes for the mentioned aluminosilicate are crystalline alkali silicates, which may exist alone or in a mixture with amorphous silicates. Alkali silicates that can be used as builders in agents according to the present invention preferably have a molar ratio of alkali oxide to SiO2 of less than 0.95, particularly 1:1.1 to 1:12, and may exist in amorphous or crystalline forms. Preferred alkali silicates are sodium silicates, particularly amorphous sodium silicates having a Na2O:SiO2 molar ratio of 1:2 to 1:2.8. Chemical formula Na2Si x O 2x+1Crystalline phyllosilicate of ·y H2O is preferably used as a crystalline silicate that may exist alone or as a mixture with amorphous silicate, wherein x is referred to as a modulus and is a number from 1.9 to 4, y is a number from 0 to 20, and preferred values ​​for x are 2, 3, or 4. Preferred crystalline phyllosilicates are those in which x is assumed to have a value of 2 or 3 in the mentioned formula. In particular, both beta-sodium and delta-sodium disilicates (Na2Si2O5·y H2O) are preferred. A substantially water-free crystalline alkali silicate of the above formula, prepared from an amorphous alkali silicate and having x as a number from 1.9 to 2.1, may also be used in the agent according to the present invention. In a further preferred embodiment of the agent according to the present invention, a crystalline sodium phyllosilicate having a modulus of 2 to 3, such as that which can be prepared from sand and soda, is used. Crystalline sodium silicate having a modulus in the range of 1.9 to 3.5 is used in a further preferred embodiment of the agent according to the present invention. Where an alkali aluminosilicate, particularly a zeolite, is also present as an additional builder, the weight ratio of aluminosilicate to silicate is preferably 1:10 to 10:1 in each case based on the anhydrous active material. In an agent containing both amorphous and crystalline alkali silicate, the weight ratio of amorphous alkali silicate to crystalline alkali silicate is preferably 1:2 to 2:1, particularly 1:1 to 2:1.

[0153] A builder is contained in the agent according to the present invention in an amount, preferably up to 60 wt.%, particularly from 5 wt.% to 40 wt.%, where desired. A water-soluble builder is particularly preferred in liquid formulations. Laundry post-treatment agents according to the present invention, such as fabric softeners, preferably do not contain inorganic builders.

[0154] Within the meaning of the present invention, polymeric thickeners are polycarboxylates that act as thickeners as polyelectrolytes, preferably homo- and copolymers of acrylic acid, in particular acrylic acid copolymers, such as acrylic acid-methacrylic acid copolymers, and polysaccharides, in particular heteropolysaccharides, and other conventional thickening polymers. Suitable polysaccharides or heteropolysaccharides are polysaccharide gums, for example gum arabic, agar, alginate, carrageenan and its salts, guar, guar gum, tragacanth, gellan, ramic acid, dextran or xanthan and their derivatives, for example propoxylated guar, and mixtures thereof. In addition to polysaccharide gums, other polysaccharide thickeners, such as starch or cellulose derivatives, for example, starch and starch derivatives of various origins, for example, hydroxyethyl starch, starch phosphate ester or starch acetate, or carboxymethyl cellulose or its sodium salt, methyl, ethyl, hydroxyethyl, hydroxypropyl, hydroxypropylmethyl or hydroxyethylmethyl cellulose or cellulose acetate may be used alternatively or preferably.

[0155] Acrylic acid polymers suitable as polymeric thickeners are, for example, high molecular weight homopolymers of acrylic acid (INCI: carbomers) cross-linked with polyalkenyl polyethers, particularly sucrose, pentaerythritol, or propylene allyl ethers, also referred to as carboxyvinyl polymers.

[0156] However, particularly suitable polymeric thickeners are the following acrylic acid copolymers: (i) acrylic acid, methacrylic acid, and preferably C 1-4(ii) copolymers of two or more monomers from the group of simple esters thereof formed using alkanols (INCI: acrylate copolymers), e.g., copolymers of methacrylic acid, butyl acrylate, and methyl methacrylate (CAS 25035-69-2) or copolymers of butyl acrylate and methyl methacrylate (CAS 25852-37-3); (ii) cross-linked high molecular weight acrylic acid copolymers, e.g., C cross-linked with an allyl ether of sucrose or pentaerythritol. 10-30 alkyl acrylates, acrylic acid, methacrylic acid, and preferably C 1-4 A copolymer of one or more monomers from the group of simple esters thereof formed using an alkanol (INCI: acrylate / C 10-30 Alkyl acrylate crosslinked polymer).

[0157] The content of the polymeric thickener is typically 8 wt.% or less, preferably 0.1 to 7 wt.%, particularly preferably 0.5 to 6 wt.%, particularly 1 to 5 wt.%, most preferably 1.5 to 4 wt.%, for example 2 to 2.5 wt.%, based on the total weight of the agent.

[0158] In particular, to stabilize the agent according to the present invention in cases where the surfactant content is high, one or more dicarboxylic acids and / or salts thereof, in particular, for example, the trade name Sokalan ® A composition of Na salts of adipic acid, succinic acid, and glutaric acid, as available as DSC, may be added. Here, based on the total weight of the detergent, it is advantageously used in an amount of 0.1 to 8 wt.%, preferably 0.5 to 7 wt.%, particularly 1.3 to 6 wt.%, and particularly preferably 2 to 4 wt.%.

[0159] However, if its use can be omitted, the agent according to the present invention preferably does not contain a dicarboxylic acid (dicarboxylic acid salt).

[0160] To determine the enhanced anti-pilling performance of the agent according to the present invention, the detergent according to the present invention may be compared with a reference detergent. A laundry system of this type may be configured as follows (all values ​​are expressed in wt.%): Reference agent: 4.4% alkylbenzenesulfonic acid, 5.6% additional anionic surfactant, 2.4% C 12 -C 18 Na salt of fatty acid (soap), 4.4% non-ionic surfactant, 0.2% phosphonate, 1.4% citric acid, 0.95% NaOH, 0.01% antifoaming agent, 2.0% glycerol, 0.08% preservative, 1% ethanol, 1.6% enzyme mixture (protease, amylase, cellulase, mannanse), and the remainder being deionized water; agent according to the present invention: 4.4% alkylbenzenesulfonic acid, 5.6% additional anionic surfactant, 2.4% C 12 -C 18 Na salt of fatty acid (soap), 4.4% non-ionic surfactant, 0.2% phosphonate, 1.4% citric acid, 0.95% NaOH, 0.01% antifoaming agent, 2.0% glycerol, 0.08% preservative, 1% ethanol, 1.6% enzyme mixture (protease, amylase, cellulase, mannanse), 0.009% polyesterase, and the remainder being deionized water. Preferably, the amount of liquid detergent used is 4.5 to 6.0 grams per liter of laundry liquid, for example, 4.7, 4.9, or 5.9 grams per liter of laundry liquid. Washing is preferably performed in a pH range of pH 8 to pH 10.5, preferably pH 8 to pH 9.

[0161] The previously mentioned embodiments of the present invention include all solid, powder, liquid, gel, or paste forms of the agent according to the present invention, which may optionally also consist of multiple phases and exist in compressed or non-compressed forms. The agent may exist as a fluid powder having a bulk density particularly of 300 g / l to 1200 g / l, particularly 500 g / l to 900 g / l, or 600 g / l to 850 g / l. Solid forms of the agent also include extruders, granules, tablets, or pouches. Alternatively, the agent may also exist in liquid, gel, or paste forms, for example, as a non-aqueous liquid detergent or non-aqueous paste, or as an aqueous liquid detergent or a water-containing paste. The agent may also exist as a single-component system. Such an agent consists of a single phase. Alternatively, the agent may also consist of multiple phases. Thus, such an agent is divided into multiple components (multi-component system).

[0162] The present invention also relates to a method for cleaning textiles, characterized by using an agent according to the present invention in at least one method step. The textile preferably contains or is made of polyester.

[0163] In various embodiments, the method described above is characterized by using the agent according to the present invention at a temperature of 0 to 100°C, preferably 0 to 80°C, more preferably 30 to 70°C, and most preferably 40 or 60°C.

[0164] This includes both manual and mechanical methods, with mechanical methods being preferred. A method for cleaning textiles is generally characterized by the fact that, in a plurality of method steps, various cleaning-active substances are added to the fabric to be cleaned and washed off after an exposure time, or the fabric to be cleaned is otherwise treated with a detergent or a solution or dilution of such agent. Since all conceivable washing or cleaning methods can be enhanced by the use of the detergent or cleaning agent according to the present invention in at least one method step, they represent embodiments of the present invention. All aspects, purposes, and embodiments described with respect to the agent according to the present invention are applicable to such objects of the present invention. Therefore, where it is stated that the present disclosure applies to the methods described above according to the present invention, the present disclosure is to be clearly referenced at the corresponding time.

[0165] Since the enzyme naturally already possesses catalytic activity and also exhibits such catalytic activity in media that do not otherwise possess cleaning power, such as simple buffers, a single and / or sole step of this method may involve contacting the stain with the polyesterase, which is the sole cleaning-active component, preferably in a buffer or water. This constitutes a further embodiment of such object of the present invention.

[0166] Alternative embodiments of such objects of the present invention are also presented as methods for processing textile raw materials or methods for managing textiles, wherein the agent according to the present invention is activated in at least one method step. Among these, a method for textile raw materials, fibers, or textiles having synthetic components, very particularly polyester, is preferred.

[0167] Furthermore, the present invention also includes the use of the agent described herein as a detergent or cleaning agent, for example as described above, for the (improved) removal of stains from textiles, particularly polyester textiles.

[0168] Finally, the present invention also relates to the use of a polyester agent, preferably for reducing the pilling effect of a detergent containing a polyester agent, preferably a detergent, particularly preferably a liquid detergent. The polyester agent is a polyester agent as defined herein. In various embodiments of use, the polyester agent is contained in the detergent in an amount of 0.00001 to 1 wt.%, preferably in an amount of 0.0001 to 0.5 wt.%, and particularly preferably in an amount of 0.001 to 0.1 wt.%. In further various embodiments, the polyester agent for reducing the pilling effect is added to a textile, particularly a textile made of polyester or containing polyester.

[0169] All aspects, purposes, and embodiments described with respect to the agent and polyester agent according to the present invention are applicable to further objects of the present invention. Therefore, where it is stated that the present disclosure applies to the method described above and the use according to the present invention, the present disclosure should be clearly referenced at the corresponding time.

[0170] Examples

[0171] Example 1: Filamentous Ascomycetes Trichoderma resei ( Trichoderma reesei Expression of polyesterase in )

[0172] For the expression of polyesterase, a synthetic gene having a nucleotide sequence suitable for the trichoderma codon method was used. The said gene was fused with various secretion signals and anchor peptides using Gibson assembly, and Escherichia coli ( Escherichia coliIt was cloned into a plasmid for amplification in ). This expression plasmid has a strong promoter for the expression of the corresponding mRNA of the polyesterase gene and additional elements that allow the selection of Escherichia coli cells having an expression construct after transformation.

[0173] From these plasmids, corresponding constructs for transformation and subsequent integration into the genome of Trichoderma lessei Not It was obtained by restricting to I. This transformed fragment contains an expression component of the polyesterase gene and a gene that enables the selection of successfully transformed cells in Trichoderma resei.

[0174] After selecting the most productive expression strain, the polyesterase was produced in sufficient quantities via fermentation to enable its use in washing application tests. The following constructs were prepared from the polyesterase, linker, and peptide anchor: SEQ ID NO:1-SEQ ID NO:6-SEQ ID NO:2

[0175] Example 2: Washing Test

[0176] Laundry matrix used

[0177] This is the commercially available detergent matrix used in the laundry test:

[0178]

[0179] Washing test to determine the anti-pilling performance of the enzyme

[0180] Twenty identical tests are performed sequentially in commercially available washing machines. Various polyester and blended textiles are used as evaluation textiles, some of which are new and some are pre-pilled. After 20 tests, the reduction of pilling in the pre-pilled fabric and the formation of pilling in the new fabric are visually evaluated.

[0181] The pre-pilled fabric is manufactured by washing it in a commercially available washing machine at 40°C for 20 repeated washing cycles.

[0182] After each washing cycle, dry the finished laundry in the dryer.

[0183] Washing conditions:

[0184] 16° dH water, clean charge 2.5 kg laundry, 40°C standard program, 50 ml of detergent as described above per machine

[0185] Amount of polyester agent tested: 50 mg of active enzyme per washing machine

[0186] Sample 1: Laundry agent alone as described above (comparative reference)

[0187] Sample 2: Laundry agent + Polyesterase 50 mg (SEQ ID NO:1)

[0188] Sample 3: Laundry agent + Polyesterase 10 mg (SEQ ID NO:1)

[0189] Sample 2: Washing agent + Polyesterase 10 mg (SEQ ID NO:1+SEQ ID NO:6+SEQ ID NO:2; i.e., polyesterase sequence SEQ ID NO:1 is connected by a peptide bond to a linker according to SEQ ID NO:6 at the C-terminus, and the linker is also connected by a peptide bond to a sequence according to SEQ ID NO:2 at the C-terminus; according to the present invention)

[0190] Result after 20 washes for the following:

[0191] A = 100% Polyester Textile, WFK30A (WFK Cleaning Technology Institute eV), New

[0192] B = 80% Cotton / 20% Polyester Textile, HS Pink Yard Good, HS Stoffe GmbH & Co. KG, Pre-pilled

[0193] Visual sampling of peel, grades 1-5, very heavily peeled = 1, not peeled = 5

[0194]

[0195] A change of 0.5 units can be considered significant.

[0196] The polyester agent according to the present invention significantly improves the appearance of the fabric compared to both a laundry agent that does not contain a polyester agent and a laundry agent that contains a polyester agent that does not have a peptide sequence having an affinity for polyester.

[0197] Example 3: Polyester-bonding strength

[0198] Using enhanced green fluorescent protein (eGFP), the affinity of various peptide sequences for polyester was tested. 100% PES fabric (WFK30A) and cotton / PES blend fabric (35% cotton / 65% polyester; WFK20A) were tested as polyesters.

[0199] The sequences tested were as follows (orientation from N-terminus to C-terminus):

[0200] Construct 1: eGFP-SEQ ID NO:6-SEQ ID NO:3

[0201] Construct 2: eGFP-SEQ ID NO:6-SEQ ID NO:4

[0202] Construct 3: eGFP-SEQ ID NO:6-SEQ ID NO:2

[0203] Construct 4: eGFP-SEQ ID NO:6-SEQ ID NO:5

[0204] The fusion protein was cloned in E. coli and prepared in a microtiter plate (MTP, 96-well), the cells were digested with lysozyme, and cell-free extracts of the individual fusion proteins were isolated.

[0205] Before the binding test, the fluorescence of the fusion protein (cell-free extract) was standardized to 100,000 RFU.

[0206] The sample textile was cut into small pieces (approximately 0.5 x 0.5 mm) and placed in individual wells of the filter plate.

[0207] 100 μl of cell-free extract was added to the textile sample using a pipette and combined for 10 minutes.

[0208] The cell-free extract was removed via a centrifugation step, and the textile sample was washed three times with buffer (100 mM Tris-HCl, pH 8.0) (3 x 5 min shaking and 5 min centrifugation).

[0209] Finally, the sample was washed with a surfactant (0.5 mM sodium dodecylbenzenesulfonate (LAS), pH 8.0) and dried (1 x 5 min shaking and 5 min centrifugation).

[0210] The samples were visually evaluated for their fluorescence using a confocal microscope (TCS SP8, Leica Microsystems CMS GmbH) (0 = no fluorescence; - / + = weak fluorescence; + = fluorescence; ++ = strong fluorescence; +++ = very strong fluorescence).

[0211] Untreated samples and samples treated only with eGFP exhibited little to no detectable fluorescence. In contrast, all samples treated with the described test constructs exhibited strong to very strong fluorescence.

[0212]

Claims

Claim 1 An agent that may be a laundry agent or a cleaning agent, characterized by containing a polyesterase covalently bonded to at least one heterogeneous peptide sequence having an affinity for polyester, wherein the peptide sequence having an affinity for polyester is selected from a peptide sequence comprising or consisting of one of the sequences given in SEQ ID NO:2 to SEQ ID NO:5 or a functional homologue thereof, wherein the functional homologue is at least 70% identical to one of the sequences given in SEQ ID NO:2 to SEQ ID NO:5, and the polyesterase comprises an amino acid sequence that is at least 95% identical to the amino acid sequence given in SEQ ID NO:1 over its entire length. Claim 2 In claim 1, an agent characterized by the following: (a) a peptide sequence having an affinity for polyester having a length of 15 to 60 amino acids; and / or (b) a peptide sequence having an affinity for polyester located at the N- and / or C-terminus with respect to a polyesterase sequence, or linked to the polyesterase sequence at the C- or N-terminus, or to the C-terminus of the polyesterase, which may be linked by a peptide linker. Claim 3 An agent according to claim 1, characterized in that a peptide sequence having an affinity for polyester is bound to a polyesterase by a linker or a peptide linker. Claim 4 In paragraph 3, the peptide linker is an agent characterized by the following: (a) having a length of 5 to 25 amino acids; and / or (b) selected from AEAAAKEAAAKEAAAKA (SEQ ID NO:6) or PPGGNRGTTTTRRPATTTGSSPGP (SEQ ID NO:7) or its homologues. Claim 5 In claim 1, an agent characterized in that the polyesterase has the following structure oriented from the N-terminus to the C-terminus: (i) polyesterase-SEQ ID NO:7-SEQ ID NO:2; (ii) polyesterase-SEQ ID NO:7-SEQ ID NO:3; (iii) polyesterase-SEQ ID NO:7-SEQ ID NO:4; (iv) polyesterase-SEQ ID NO:7-SEQ ID NO:5; (v) polyesterase-SEQ ID NO:6-SEQ ID NO:2; (vi) polyesterase-SEQ ID NO:6-SEQ ID NO:3; (vii) polyesterase-SEQ ID NO:6-SEQ ID NO:4; or (viii) polyesterase-SEQ ID NO:6-SEQ ID NO:

5. Claim 6 An agent according to claim 1, characterized in that the polyesterase contains an amino acid sequence identical to the amino acid sequence given in SEQ ID NO:1 over its entire length. Claim 7 An agent according to claim 1, wherein the polyester agent is contained in the agent in an amount of 0.00001 to 1 wt.%, or in an amount of 0.0001 to 0.5 wt.%, or in an amount of 0.001 to 0.1 wt.%. Claim 8 A method for cleaning textiles, characterized by using an agent according to any one of claims 1 to 7 in at least one method step. Claim 9 A method according to claim 8, characterized in that the textile contains polyester or is made of polyester. Claim 10 An agent for use in stain removal in any one of claims 1 to 7, wherein the stain removal may be stain removal from polyester-containing textiles. Claim 11 A composition comprising a polyesterase covalently bonded to at least one heteropeptide sequence having an affinity for polyester, for use in reducing the peeling effect of an agent, a detergent, or a liquid detergent and / or enhancing the anti-graying effect, wherein the agent, the detergent, or the liquid detergent contains the polyesterase, the polyesterase is as defined in any one of claims 1 to 6, and the peptide sequence having an affinity for polyester is as defined in any one of claims 1 to 6. Claim 12 A composition according to claim 11, wherein the polyester agent is contained in the agent in an amount as defined in claim 7. Claim 13 delete Claim 14 delete

Images