Process for the production of (1r,2s)-2,6-dimethyl-1-indanamine using dynamic kinetic stereoisomer resolution

Abstract

Described is a process for producing nearly enantiomerically pure (1R,2S)-2,6-dimethyl-1-indanamine, said process being characterized by the reaction of a mixture of the four stereoisomers of 2,6-dimethyl-1-indanamine with an acylating or carboxylating agent in the presence of a protein with lipase activity and being carried out using dynamic kinetic stereoisomer resolution.

Description

[0001]BCS221003-Ausland Dr. ML 1 Bayer AG Process for the preparation of (1R,2S)-2,6-dimethyl-1-indanamine by dynamic-kinetic stereoisomer resolution Description The present invention relates to a process for the preparation of virtually enantiomerically pure (1R,2S)-2,6-dimethyl-1-indanamine, which is a valuable building block for the synthesis of the herbicidal active ingredient indaziflam. In particular, the present invention relates to a process for the preparation of virtually enantiomerically pure (1R,2S)-2,6-dimethyl-1-indanamine by enzyme-catalyzed, stereoselective acylation of racemic 2,6-dimethyl-1-indanamine. From the state of the art, only processes for the preparation of almost enantiomerically pure (1R,2S)-2,6-dimethyl-1-indanamine are known, which are only suitable for laboratory scale but not for industrial use due to the use of expensive reaction components and catalysts.WO 2004 / 69814 A1 discloses a process characterized by the following five reaction steps: 1. Preparation of a mixture of the four stereoisomers of 2,6-dimethyl-1-aminoindane by palladium-catalyzed reduction of the corresponding oxime. 2. Column chromatographic separation of these four stereoisomers into the cis and trans isomers. 3. Reaction of the trans isomers with methyl 2-methoxyacetate in the presence of the enzyme Novozym 435® to form the corresponding acetylated (1R,2S)-2,6-dimethyl-1-indaneamine. 4. Isolation of the acetylated (1R,2S)-2,6-dimethyl-1-indaneamine. 5. Acidic hydrolysis of the acetylated (1R,2S)-2,6-dimethyl-1-indaneamine to form the free (1R,2S)-2,6-dimethyl-1-indaneamine. Tetrahedron 2007, 63 (29), 6755-6763 describes a process for the preparation of (1R,2S)-2,6-dimethyl-1-indanamine, which is characterized by the following two reaction steps: 1.Racemic 1,6-dimethylindan-1-one is reduced diastereoselectively (96:4 dr) and enantioselectively (98:2 er) using a chiral ruthenium catalyst to the corresponding (1S,2S)-2,6-dimethylindan-1-ol in 80% yield. 2. Treatment of the resulting (1S,2S)-2,6-dimethylindan-1-ol with diphenylphosphoryl azide and subsequent reduction with lithium aluminum hydride leads to (1R,2S)-2,6-dimethyl-1-indanamine in 76% yield. BCS221003-Ausland Dr. ML 2 A disadvantage of this process, in addition to the use of expensive reagents, is the long reaction time of eight days. The object of the present invention was to provide a process for the preparation of almost enantiomerically pure (1R,2S)-2,6-dimethyl-1-indanamine, which overcomes the disadvantages known from the prior art.A process for the preparation of almost enantiomerically pure (1R,2S)-2,6-dimethyl-1-indanamine has been found, which is characterized a) by selective reaction of a mixture of the four stereoisomers of 2,6-dimethyl-1-indanamine with an acylating or carboxylating agent in the presence of a protein with the activity of a lipase, and b) by a continuous metal-catalyzed isomerization of the mixture of three undesired stereoisomers of 2,6-dimethyl-1-indanamine to the four stereoisomers of 2,6-dimethyl-1-indanamine. The present invention relates to a process for the preparation of almost enantiomerically pure (1R,2S)-2,6-dimethyl-1-indanamine, characterized in that 1. in a first step a mixture of the four stereoisomers of 2,6-dimethyl-1-indanamine (I) is reacted with an acylating or carboxylating agent RC(=O)R. 1a) in the presence of a protein with the activity of a lipase, it is selectively converted to the corresponding amide or carbamate (II) and a mixture (III) of the unreacted stereoisomers of 2,6-dimethyl-1-indanamine, and b) the mixture (III) is isomerized in the presence of a metal catalyst and under hydrogen pressure at the same time as the biocatalytic conversion to the starting material of the four stereoisomers of 2,6-dimethyl-1-indanamine (I): M etallkatalysator, wherein the protein is encoded by an amino acid sequence selected from the group consisting of BCS221003-Ausland Dr. ML 3 I. Proteins which have at least 80%, preferably 85%, more preferably 90%, even more preferably 95%, even more preferably 96%, even more preferably 97%, particularly preferably 98%, most preferably 99% identity to the amino acid sequence shown under SEQ ID No.1, II. Proteins which have at least 80%, preferably 85%, more preferably 90%, even more preferably 95%, even more preferably 96%, even more preferably 97%, particularly preferably 98%, most preferably 99% identity to the amino acid sequence shown under SEQ ID No.1, apart from the fact that the amino acid sequence which has at least 80%, preferably 85%, more preferably 90%, even more preferably 95%, even more preferably 96%, even more preferably 97%, particularly preferably 98%, most preferably 99% identity to the amino acid sequence shown under SEQ ID No.11, has a modification selected from the group consisting of i. the amino acid at position 186 is different from L; ii. the amino acid at position 280 is different from L; iii. the amino acid at position 312 is different from P; iv. the amino acid at position 3 is different from M; v. the amino acid at position 29 is different from N; vi. the amino acid at position 17 is different from L; vii. the amino acid at position 4 is different from S; viii. the amino acid at position 18 is different from V; ix. the amino acid at position 202 is different from A; x. the amino acid at position 301 is different from D; xi. the amino acid at position 309 is different from P; xii. the amino acid at position 31 is different from Q; xiii. the amino acid at position 111 is different from Q; xiv. the amino acid at position 85 is different from W; xv. the amino acid at position 8 is different from K; xvi.the amino acid at position 79 is different from E; xvii. the amino acid at position 40 is different from K; 2. that in a second step the amide or carbamate (II) is separated from the minor components by crystallization, and BCS221003-Ausland Dr. ML 4 3. that in a third step the amide or carbamate (II) is converted into the almost enantiomerically pure (1R,2S)-2,6-dimethyl-1-indanamine (IV) using a base or an acid. where R is a radical from the group CH2OCH3, CH2OCH2CH3, CH3, OCH3, OCH2CH3, OCH(CH3) 2, OCH2CH2CH2CH3, OCH2CHCH2 and OCH2(C6H5), and wherein R 1a radical from the group OCH3, OCH2CH3, OCH(CH3)2, OCH2CH2CH2CH3, OCH2CHCH2, and OCH2(C6H5). The mixture of the four stereoisomers of 2,6-dimethyl-1-indanamine (I) required as starting material for the process according to the invention is known and can be prepared, for example, as described in WO2004 / 69814. Preferably, R is a radical from the group OCH3 and OCH2CH3, and R 1means a residue from the group OCH3 and OCH2CH3. Proteins with the activity of a lipolytic enzyme or a lipase are used in the process according to the invention. SEQ ID No. 1 represents the amino acid sequence of a wild-type lipolytic protein. The wild-type lipase is derived from an uncultivated bacterium from an environmental sample, which can be obtained from GenPept (PDB) under accession no. QRD81023 (version ORD81023.1). In case of doubt between the amino acid sequence shown in SEQ ID No. 1 and the sequence shown in the above database entry, SEQ ID No. 1 takes precedence. Also described are proteins with the activity of a lipolytic enzyme or a lipase, wherein the amino acid sequences of these proteins represent variants of a known protein.which has the activity of a lipolytic enzyme or a lipase. In particular, the amino acid sequences of proteins with the activity of a lipase described herein BCS221003-Ausland Dr. ML 5 represent variants of the amino acid sequence represented by the amino acid shown in SEQ ID No. 1, wherein in the amino acid sequence shown under SEQ ID No. 1 at least the amino acid at position 186, the amino acid at position 280, the amino acid at position 312, the amino acid at position 3, the amino acid at position 29, the amino acid at position 17, the amino acid at position 4, the amino acid at position 18, the amino acid at position 202, the amino acid at position 301, the amino acid at position 309, the amino acid at position 31, the amino acid at position 111, the amino acid at position 85, the amino acid at position 8, the amino acid at position 79 or the amino acid at position 40 is different from the amino acid,which is present at the corresponding amino acid position in the sequence shown under SEQ ID No. 1. The term "variant" as used here means an object that differs from an object known in the art. With regard to nucleic acid molecules and proteins, variants are understood to mean a nucleic acid sequence or an amino acid sequence that deviates from corresponding known sequences, but which encodes a protein that has the same function or catalyzes the same reaction, e.g., the function of encoding a protein with lipase activity. "Deviation" of nucleic acid molecule sequences and amino acid sequences from known nucleic acid sequences and protein sequences meansthat the sequences comprise substitutions (replacements) and / or deletions and / or insertions of nucleotides or amino acids compared to the corresponding known nucleic acid sequences or amino acid sequences. Proteins with the activity of a lipase are usually used, wherein the proteins are encoded by an amino acid sequence which has at least 80%, preferably 85%, more preferably 90%, even more preferably 95%, even more preferably 96%, even more preferably 97%, particularly preferably 98%, most preferably 99% identity to the amino acid sequence shown under SEQ ID No. 1. Proteins with the activity of a lipase are also used, wherein the proteins are encoded by an amino acid sequence which has at least 80%, preferably 85%, more preferably 90%, even more preferably 95%, even more preferably 96%, even more preferably 97%, particularly preferably 98%.most preferably has 99% identity to the amino acid sequence shown under SEQ ID No. 1, apart from the fact that the amino acid sequence which has at least 80%, preferably 85%, more preferably 90%, even more preferably 95%, even more preferably 96%, even more preferably 97%, particularly preferably 98%, most preferably 99% identity to the amino acid sequence shown under SEQ ID No. 1, has a modification selected from the group consisting of BCS221003-Ausland Dr. ML 6 i. the amino acid at position 186 is different from L, preferably the amino acid at position 186 is F, W, Y, E, D, Q, T, H, P, C, K, S, N, I or V, more preferably the amino acid at position 186 is F, W, Y, E, D or K, particularly preferably the amino acid at position 186 is W or Y, most preferably the amino acid at position 186 is Y; ii. the amino acid at position 280 is different from L, preferably the amino acid at position 280 is E, S, K, D or A,more preferably, the amino acid at position 280 is A; iii. the amino acid at position 312 is different from P, preferably the amino acid at position 312 is N, F, D, Q or K, more preferably the amino acid at position 312 is N; iv. the amino acid at position 3 is different from M, preferably the amino acid at position 3 is L, Q or C, more preferably the amino acid at position 3 is Q; v. the amino acid at position 29 is different from N, preferably the amino acid at position 29 is H, W or Y, more preferably the amino acid at position 29 is H or W, most preferably the amino acid at position 29 is H; vi. the amino acid at position 17 is different from L, preferably the amino acid at position 17 is P or T, more preferably the amino acid at position 17 is P; vii. the amino acid at position 4 is different from S, preferably the amino acid at position 4 is P or L,more preferably, the amino acid at position 4 is P; viii. the amino acid at position 18 is different from V, preferably the amino acid at position 18 is A, T, C or S, more preferably the amino acid at position 18 is A or C; ix. the amino acid at position 202 is different from A, preferably the amino acid at position 202 is Q or N. More preferably, the amino acid at position 202 is N; x. the amino acid at position 301 is different from D, preferably the amino acid at position 301 is A; BCS221003-Ausland Dr. ML 7 xi. the amino acid at position 309 is different from P, preferably the amino acid at position 309 is C; xii. the amino acid at position 31 is different from Q, preferably the amino acid at position 31 is W; xiii. the amino acid at position 111 is different from Q, preferably the amino acid at position 111 is E; xiv. the amino acid at position 85 is different from W,Preferably, the amino acid at position 85 is H; xv. the amino acid at position 8 is different from K, preferably the amino acid at position 8 is E; xvi. the amino acid at position 79 is different from K, preferably the amino acid at position 79 is S, I or W, more preferably the amino acid at position 79 is S; xvii. the amino acid at position 40 is different from K, preferably the amino acid at position 40 is M. The meaning of the amino acid abbreviations A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, TV, W, Y can be derived hereinafter in Table 2 in the section entitled "Description of the Sequences". A further embodiment of the invention relates to proteins with the activity of a lipase, wherein the proteins are selected from the group consisting of a) proteins comprising the amino acid sequence shown in SEQ ID No. 1, apart fromthat the amino acid at position 186 is different from L; b) proteins with an amino acid sequence with at least 80%, preferably 85%, more preferably 90%, even more preferably 95%, even more preferably 96%, even more preferably 97%, particularly preferably 98%, most preferably 99% identity to the amino acid sequence shown under a), provided that the amino acid at position 186 is different from L. Preferably, the amino acid in the proteins according to a) or b) at position 186 is F, W, Y, E, D, Q, T, H, P, C, K, S, N, I or V. More preferably, the amino acid at position 186 is F, W, Y, E, D or BCS221003-Ausland Dr. ML 8 K. Particularly preferred is the amino acid at position 186 W or Y. Most preferred is the amino acid at position 186 Y. An “amino acid corresponding to position X” in a first amino acid sequence (e.g. position 3 in SEQ ID No. 1) means herein that an amino acid of a second amino acid sequence,compared to the first amino acid sequence, appears at position x of the first amino acid sequence in a pairwise sequence alignment of the first amino acid sequence with the second amino acid sequence, if the numbering of the amino acids of the second amino acid sequence differs from the amino acid numbering of the first amino acid sequence. In the context of the present invention, the term "identity" in relation to sequence identity or identical sequences is understood to mean the number of identical amino acids or nucleotides that a first nucleic or amino acid sequence has in common with another (second) nucleic or amino acid sequence over the entire sequence length, expressed as a percentage. "Sequence identity" can be determined by aligning two amino acid or two nucleotide sequences using global or local alignment algorithms,as included, for example, in well-known software such as GAP or BESTFIT or the Emboss program "Needle." This software uses the global alignment algorithm of Needleman and Wunsch to align two sequences across their entire length, maximizing the number of matches and minimizing the number of gaps. Generally, the default parameters are used, with a gap creation penalty of 10 and a gap extension penalty of 0.5 (for both nucleotide and protein alignment). For nucleotides, the default scoring matrix used is DNAFULL, and for proteins, the default scoring matrix is ​​Blosum62 (Henikoff & Henikoff, 1992, PNAS 89, 10915–10919). Sequence alignments and percentage sequence identity scores can be determined, for example, using software such as EMBOSS.available on the EBI website (ebi.ac.uk / Tools / emboss / ). Alternatively, sequence similarity or identity can be determined by searching databases (e.g., EMBL, GenBank) using well-known algorithms and output formats such as FASTA, BLAST, etc., but preferably, hits should be retrieved and pairwise aligned to conclusively determine sequence identity. If sequences to be compared are of different lengths, identity should be determined by determining the identity as a percentage of the number of amino acids or nucleotides that the shorter sequence has in common with the longer sequence. Identity is preferably determined using the well-known and publicly available computer program ClustalW (Thompson et al., Nucleic Acids Research 22 (1994), 4673–4680). ClustalW is publicly available from Julie Thompson (Thompson@EMBL-Heidelberg.DE) and Toby Gibson (Gibson@EMBL-Heidelberg.DE),European BCS221003-Ausland Dr. ML 9 Molecular Biology Laboratory, Meyerhofstraße 1, D 69117 Heidelberg, Germany. ClustalW can also be downloaded from various websites, such as IGBMC (Institut de Génétique et de Biologie Moléculaire et Cellulaire, BP163, 67404 Illkirch Cedex, France; ftp: / / ftp-igbmc.u-strasbg.fr / pub / ) and EBI (ftp: / / ftp.ebi.ac.uk / pub / software / ), as well as from the mirrored websites of EBI (European Bioinformatics Institute, Wellcome Trust Genome Campus, Hinxton, Cambridge CB101SD, UK). Preferably, the ClustalW computer program version 1.8 is used to determine the identity between proteins described in the context of the present invention and other proteins. The parameters must be set as follows: KTUPLE=1, TOPDIAG=5, WINDOW=5, PAIRGAP=3, GAPOPEN=10, GAPEXTEND=0.05, GAPDIST=8, MAXDIV=40, MATRIX=GONNET, ENDGAPS(OFF), NOPGAP,NOHGAP. Preferably, the computer program ClustalW version 1.8 is used to determine the identity, for example, between the nucleotide sequence of the nucleic acid molecules described in connection with the present invention and the nucleotide sequence of other nucleic acid molecules. The parameters must be set as follows: KTUPLE=2, TOPDIAGS=4, PAIRGAP=5, DNAMATRIX:IUB, GAPOPEN=10, GAPEXT=5, MAXDIV=40, TRANSITIONS: unweighted. "Identity" further means that there is a functional and / or structural equivalence between the nucleic acid molecules in question or the proteins they encode. Functional equivalence means that the nucleic acid molecule sequences or the amino acid sequences encode a protein with lipase activity. The nucleic acid molecules that are homologous to the molecules described above and are derivatives of these molecules are generally variants of these molecules.represent modifications that have the same biological function or catalyze the same reaction, i.e., encode a protein with lipase activity. These can be either naturally occurring variants, for example, sequences from other species, or mutations, whereby these mutations may have occurred naturally or been introduced by targeted mutagenesis. In addition, the variants can be synthetically produced sequences. The allelic variants can be either naturally occurring variants or synthetically produced variants or variants generated by recombinant DNA techniques. However, what is crucial for the present invention is that these variants encode proteins with lipase activity and the amino acid substitutions (replacements) described here,deletions or insertions relating to the proteins according to the invention. BCS221003-Ausland Dr. ML 10 A special type of derivative is, for example, nucleic acid molecules which differ from the nucleic acid molecules described in the present invention due to the degeneracy of the genetic code. According to NC-IUBMB (Nomenclature Committee of the International Union of Biochemistry and Molecular Biology), lipases belong to the class of hydrolases (EC 3). Hydrolase is a class of enzymes which usually act as biochemical catalysts and use water to break a chemical bond, which typically leads to the division of a larger molecule into smaller molecules. The group of hydrolases includes enzymes which act on ester bonds (EC 3.1), such as carboxylic acid ester hydrolases (EC 3.1.1) and, as a subgroup, lipases (EC 3.1.1.3). Lipases have been identified from plants, mammals and microorganisms, e.g. B. Pseudomonas,Vibrio, Acinetobacter, Burkholderia, Chromobacterium, cutinase from Fusarium solani (FSC), Candida antarctica A (CalA), Rhizopus oryzae (ROL), Thermomyces lanuginosus (TLL), Rhizomucor miehei (RML), Aspergillus niger, Fusarium heterosporum, Fusarium oxysporum, or Fusarium culmorum. If a protein exhibits lipase activity, this can be detected using methods known and described in the art. It is not critical which method is used to detect whether a protein according to the invention exhibits lipase activity. Preferably, in connection with the present invention, the method is described in the "Example" section. Lipase variant proteins according to the invention can contain further amino acid modifications (amino acid substitutions,-deletions or insertions) compared to the amino acid sequences described above with respect to the amino acid sequence shown under SEQ ID No. 1. In addition to the lipase variants described above under point a) with respect to the amino acid sequences shown under SEQ ID No. 1, at least one, two, three, four, five, six, seven further amino acid substitutions at positions 79, 202, 280, 301, 3, 11, 17, 40 or 111 can be present. In other words, the protein according to the invention with the activity of a lipase is selected from the group consisting of a) proteins which comprise the amino acid sequence shown in SEQ ID No. 1, except that the amino acid at position 186 is different from L and that they have at least one, two, three, four, five, six, seven or more further amino acid substitutions,selected from the group consisting of (i) the amino acid at position 79 is different from E; (ii) the amino acid at position 202 is different from A; (iii) the amino acid at position 280 is different from L; (iv) the amino acid at position 301 is different from D; (v) the amino acid at position 3 is different from M; (vi) the amino acid at position 11 is different from C; (vii) the amino acid at position 17 is different from L; (viii) the amino acid at position 40 is different from K; (ix) the amino acid at position 111 is different from Q; and b) proteins having an amino acid sequence with at least 80%, preferably 85%, more preferably 90%, even more preferably 95%, even more preferably 96%, even more preferably 97%, particularly preferably 98%, most preferably 99% identity to the amino acid sequence shown directly above under a),provided that the amino acid at position 186 is different from L and with at least one further amino acid substitution selected from groups (i) to (ix) mentioned directly above. Preferably, the amino acid at position 79 is S, W or I,more preferably S; preferably the amino acid at position 202 is N; preferably the amino acid at position 280 is A; preferably the amino acid at position 301 is A; preferably the amino acid at position 3 is Q; preferably the amino acid at position 11 is A; preferably the amino acid at position 17 is P; preferably the amino acid at position 40 is M; preferably the amino acid at position 111 is E. Furthermore, lipase variant proteins according to the invention may, apart from the further amino acid modifications, have additional amino acid substitutions compared to the amino acid sequences described hereinabove with respect to the amino acid sequence shown under SEQ ID No. 1. These additional amino acid substitutions refer to positions of the amino acid sequence that are different from position(s) 79, 202, 280, 301, 3, 11, 17, 40 or 111.which relate to the further amino acid modifications. The lipase variants described above under point a) with respect to the amino acid sequences shown under SEQ ID No. 1 may have at least one, two, three, four, five, six, seven or more additional amino acid substitutions at positions 4, 8, 18, 29, 31, 42, 84, 85, 192, 217, 309 or 312. The amino acid at position 4 is different from S, preferably the amino acid at this position is P. The amino acid at position 8 is different from K, preferably the amino acid at this position is E. The amino acid at position 18 is different from V, preferably the amino acid at this position is C. The amino acid at position 29 is different from N, preferably the amino acid at this position is W or H. The amino acid at position 31 is different from Q, preferably the amino acid at this position is W. The amino acid at position 42 is different from L,Preferably, the amino acid at this position is D. The amino acid at position 84 is different from N, preferably the amino acid at this position is T. The amino acid at position 85 is different from W, preferably the amino acid at this position is H. The amino acid at position 192 is different from F, preferably the amino acid at this position is A or V. The amino acid at position 217 is different from Q, preferably the amino acid at this position is M. The amino acid at position 309 is different from P, preferably the amino acid at this position is C. The amino acid at position 312 is different from P, preferably the amino acid at this position is N. BCS221003-Ausland Dr. ML 12 A further embodiment of the invention accordingly relates to proteins according to the invention which have further amino acid modifications, preferably these embodiments are proteins with the activity of a lipase,wherein the proteins are selected from the group consisting of: proteins which have the amino acid sequence shown in SEQ ID No. 1, with the exception that ^ ^ the amino acid at position 186 is different from L and the amino acid at position 79 is different from E, wherein it is preferred that the amino acid at position 186 is preferably W or Y, more preferably Y, and that the amino acid at position 79 is preferably S, W or I, more preferably S; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 202 is different from A, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 202 is preferably N; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 280 is different from L, wherein it is preferred that the amino acid at position 186 is preferably W or Y,and that the amino acid at position 280 is preferably A; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 301 is different from D, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 301 is preferably A; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 3 is different from M, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 3 is preferably Q; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 11 is different from C, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 11 is preferably A; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 17 is different from L, wherein it is preferred,that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 17 is preferably P; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 40 is different from K, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 40 is preferably M; BCS221003-Ausland Dr. ML 13 ^ ^ the amino acid at position 186 is different from L and the amino acid at position 111 is different from Q, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 111 is preferably E; ^ ^ Proteins with an amino acid sequence that is at least 80%, preferably 85%, more preferably 90%, even more preferably 95%, even more preferably 96%, even more preferably 97%, particularly preferably 98%, most preferably 99% identical to the amino acid sequence shown under a),provided that the amino acid at position 186 is different from L, it is preferred that the amino acid at position 186 is preferably W or Y, and wherein the proteins have at least one further amino acid substitution selected from the group shown under the listed symbols directly above. Preferably, the lipase variants described herein above under point a) with respect to the amino acid sequences shown under SEQ ID No. 1 can have at least two further amino acid substitutions at positions 79, 202, 280, 301, 3, 11, 17, 40 or 111. Preferably, the amino acid at position 79 is S, W or I,more preferably S; preferably the amino acid at position 202 is N; preferably the amino acid at position 280 is A; preferably the amino acid at position 301 is A; preferably the amino acid at position 3 is Q; preferably the amino acid at position 11 is A; preferably the amino acid at position 17 is P; preferably the amino acid at position 40 is M; preferably the amino acid at position 111 is E. A further embodiment of the invention accordingly relates to proteins according to the invention which have further amino acid modifications, preferably these embodiments are proteins with the activity of a lipase, wherein the proteins are selected from the group consisting of proteins with the amino acid sequence shown in SEQ ID No. 1, with the exception that ^ ^ the amino acid at position 186 is different from L and the amino acid at position 79 is different from E and the amino acid at position 202 is different from A, wherein it is preferredthat the amino acid at position 186 is preferably W or Y, and that the amino acid at position 79 is preferably S, W or I, more preferably S, and that the amino acid at position 202 is preferably N; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 79 is different from E and the amino acid at position 280 is different from L, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 79 is preferably S, W or I, more preferably S, and that the amino acid at position 280 is preferably A; BCS221003-Ausland Dr. ML 14 ^ ^ the amino acid at position 186 is different from L and the amino acid at position 79 is different from E and the amino acid at position 301 is different from D, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 79 is preferably S, W or I, more preferably S,and that the amino acid at position 301 is preferably A; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 79 is different from E and the amino acid at position 3 is different from M, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 79 is preferably S, W or I, more preferably S, and that the amino acid at position 3 is preferably Q; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 79 is different from E and the amino acid at position 11 is different from C, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 79 is preferably S, W or I, more preferably S,and that the amino acid at position 11 is preferably A; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 79 is different from E and the amino acid at position 17 is different from L, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 79 is preferably S, W or I, more preferably S, and that the amino acid at position 17 is preferably P; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 79 is different from E and the amino acid at position 40 is different from K, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 79 is preferably S, W or I, more preferably S,and that the amino acid at position 40 is preferably M; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 79 is different from E and the amino acid at position 111 is different from Q, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 79 is preferably S, W or I, more preferably S, and that the amino acid at position 111 is preferably E; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 202 is different from A and the amino acid at position 280 is different from L, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the BCS221003-Ausland Dr. ML 15 amino acid at position 202 is preferably N,and that the amino acid at position 280 is preferably A; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 202 is different from A and the amino acid at position 301 is different from D, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 202 is preferably N, and that the amino acid at position 301 is preferably A; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 202 is different from A and the amino acid at position 3 is different from M, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 202 is preferably N, and that the amino acid at position 3 is preferably Q; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 202 is different from A and the amino acid at position 11 is different from C,wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 202 is preferably N, and that the amino acid at position 11 is preferably A; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 202 is different from A and the amino acid at position 17 is different from L, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 202 is preferably N, and that the amino acid at position 17 is preferably P; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 202 is different from A and the amino acid at position 40 is different from K, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 202 is preferably N,and that the amino acid at position 40 is preferably M; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 202 is different from A and the amino acid at position 111 is different from Q, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 202 is preferably N, and that the amino acid at position 111 is preferably E; BCS221003-Ausland Dr. ML 16 ^ ^ the amino acid at position 186 is different from L and the amino acid at position 280 is different from L and the amino acid at position 301 is different from D, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 280 is preferably A,and that the amino acid at position 301 is preferably A; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 280 is different from L and the amino acid at position 3 is different from M, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 280 is preferably A, and that the amino acid at position 3 is preferably Q; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 280 is different from L and the amino acid at position 11 is different from C, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 280 is preferably A, and that the amino acid at position 11 is preferably A; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 280 is different from L and the amino acid at position 17 is different from L, wherein it is preferred,that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 280 is preferably A, and that the amino acid at position 17 is preferably P; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 280 is different from L and the amino acid at position 40 is different from K, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 280 is preferably A, and that the amino acid at position 40 is preferably M; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 280 is different from L and the amino acid at position 111 is different from Q, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 280 is preferably A,and that the amino acid at position 111 is preferably E; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 301 is different from D and the amino acid at position 3 is different from M, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the BCS221003-Ausland Dr. ML 17 amino acid at position 301 is preferably A, and that the amino acid at position 3 is preferably Q; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 301 is different from D and the amino acid at position 11 is different from C, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 301 is preferably A,and that the amino acid at position 11 is preferably A; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 301 is different from D and the amino acid at position 17 is different from L, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 301 is preferably A, and that the amino acid at position 17 is preferably P; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 301 is different from D and the amino acid at position 40 is different from K, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 301 is preferably A, and that the amino acid at position 40 is preferably M; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 301 is different from D and the amino acid at position 111 is different from Q,wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 301 is preferably A, and that the amino acid at position 111 is preferably E; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 3 is different from M and the amino acid at position 11 is different from C, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 3 is preferably Q, and that the amino acid at position 11 is preferably A; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 3 is different from M and the amino acid at position 17 is different from L, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 3 is preferably Q,and that the amino acid at position 17 is preferably P; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 3 is different from M and the amino acid at position 40 is different from K, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 3 is preferably Q, and that the amino acid at position 40 is preferably M; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 3 is different from M and the amino acid at position 111 is different from Q, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 3 is preferably Q, and that the amino acid at position 111 is preferably E; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 11 is different from C and the amino acid at position 17 is different from L,wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 11 is preferably A, and that the amino acid at position 17 is preferably P; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 11 is different from C and the amino acid at position 40 is different from K, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 11 is preferably A, and that the amino acid at position 40 is preferably M; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 11 is different from C and the amino acid at position 111 is different from Q, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 11 is preferably A,and that the amino acid at position 111 is preferably E; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 17 is different from L and the amino acid at position 40 is different from K, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 17 is preferably P, and that the amino acid at position 40 is preferably M; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 17 is different from L and the amino acid at position 111 is different from Q, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 17 is preferably P,and that the amino acid at position 111 is preferably E; BCS221003-Ausland Dr. ML 19 ^ ^ the amino acid at position 186 is different from L and the amino acid at position 40 is different from K and the amino acid at position 111 is different from Q, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 40 is preferably M, and that the amino acid at position 111 is preferably E; ^ ^ Proteins with an amino acid sequence which is at least 80%, preferably 85%, more preferably 90%, even more preferably 95%, even more preferably 96%, even more preferably 97%, particularly preferably 98%, most preferably 99% identical to the amino acid sequence shown under a), provided that the amino acid at position 186 is different from L, wherein it is preferred that the amino acid at position 186 is preferably W or Y,and wherein the proteins have at least two further amino acid substitutions selected from the group shown under the listed symbols directly above. Preferably, the lipase variants described herein above under point a) with respect to the amino acid sequences shown under SEQ ID No. 1 may have at least three further amino acid substitutions at positions 79, 202, 280, 301, 3, 11, 17, 40, or 111. Preferably, the amino acid at position 79 is S, W, or I.more preferably S; preferably the amino acid at position 202 is N; preferably the amino acid at position 280 is A; preferably the amino acid at position 301 is A; preferably the amino acid at position 3 is Q; preferably the amino acid at position 11 is A; preferably the amino acid at position 17 is P; preferably the amino acid at position 40 is M; preferably the amino acid at position 111 is E. A further embodiment of the invention accordingly relates to proteins according to the invention which have further amino acid modifications, preferably these embodiments are proteins with the activity of a lipase, wherein the proteins are selected from the group consisting of proteins with the amino acid sequence shown in SEQ ID No. 1, with the exception,that ^ ^ the amino acid at position 186 is different from L and the amino acid at position 79 is different from E and the amino acid at position 202 is different from A and the amino acid at position 280 is different from L, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 79 is preferably S, W or I, more preferably S, and that the amino acid at position 202 is preferably N, and that the amino acid at position 280 is preferably A; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 79 is different from E and the amino acid at position 202 is different from A and the amino acid at position 301 is different from D, wherein it is preferred that the amino acid at position BCS221003-Ausland Dr. ML 20 186 is preferably W or Y, and that the amino acid at position 79 is preferably S, W or I, more preferably S,and that the amino acid at position 202 is preferably N, and that the amino acid at position 301 is preferably A; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 79 is different from E and the amino acid at position 202 is different from A and the amino acid at position 3 is different from M, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 79 is preferably S, W or I, more preferably S, and that the amino acid at position 202 is preferably N, and that the amino acid at position 3 is preferably Q; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 79 is different from E and the amino acid at position 202 is different from A and the amino acid at position 11 is different from C, wherein it is preferred that the amino acid at position 186 is preferably W or Y,and that the amino acid at position 79 is preferably S, W or I, more preferably S, and that the amino acid at position 202 is preferably N, and that the amino acid at position 11 is preferably A; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 79 is different from E and the amino acid at position 202 is different from A and the amino acid at position 17 is different from L, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 79 is preferably S, W or I, more preferably S, and that the amino acid at position 202 is preferably N, and that the amino acid at position 17 is preferably P; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 79 is different from E and the amino acid at position 202 is different from A and the amino acid at position 40 is different from K, wherein it is preferred,that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 79 is preferably S, W or I, more preferably S, and that the amino acid at position 202 is preferably N, and that the amino acid at position 40 is preferably M; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 79 is different from E and the amino acid at position 202 is different from A and the amino acid at position 111 is different from Q, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 79 is preferably S, W or I, more preferably S, and that the amino acid at position 202 is preferably N,and that the amino acid at position 111 is preferably E; BCS221003-Ausland Dr. ML 21 ^ ^ the amino acid at position 186 is different from L and the amino acid at position 79 is different from E and the amino acid at position 280 is different from L and the amino acid at position 301 is different from D, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 79 is preferably S, W or I, more preferably S, and that the amino acid at position 280 is preferably A, and that the amino acid at position 301 is preferably A; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 79 is different from E and the amino acid at position 280 is different from L and the amino acid at position 3 is different from M, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 79 is preferably S, W or I,more preferably S, and that the amino acid at position 280 is preferably A, and that the amino acid at position 3 is preferably Q; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 79 is different from E and the amino acid at position 280 is different from L and the amino acid at position 11 is different from C, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 79 is preferably S, W or I, more preferably S, and that the amino acid at position 280 is preferably A, and that the amino acid at position 11 is preferably A; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 79 is different from E and the amino acid at position 280 is different from L and the amino acid at position 17 is different from L, wherein it is preferred that the amino acid at position 186 is preferably W or Y,and that the amino acid at position 79 is preferably S, W or I, more preferably S, and that the amino acid at position 280 is preferably A, and that the amino acid at position 17 is preferably P; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 79 is different from E and the amino acid at position 280 is different from L and the amino acid at position 40 is different from K, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 79 is preferably S, W or I, more preferably S, and that the amino acid at position 280 is preferably A, and that the amino acid at position 40 is preferably M; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 79 is different from E and the amino acid at position 280 is different from L and the amino acid at position 111 is different from Q, wherein it is preferredthat the amino acid at position BCS221003-Ausland Dr. ML 22 186 is preferably W or Y, and that the amino acid at position 79 is preferably S, W or I, more preferably S, and that the amino acid at position 280 is preferably A, and that the amino acid at position 111 is preferably E; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 79 is different from E and the amino acid at position 301 is different from D and the amino acid at position 3 is different from M, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 79 is preferably S, W or I, more preferably S, and that the amino acid at position 301 is preferably A,and that the amino acid at position 3 is preferably Q; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 79 is different from E and the amino acid at position 301 is different from D and the amino acid at position 11 is different from C, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 79 is preferably S, W or I, more preferably S, and that the amino acid at position 301 is preferably A, and that the amino acid at position 11 is preferably A; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 79 is different from E and the amino acid at position 301 is different from D and the amino acid at position 17 is different from L, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 79 is preferably S, W or I, more preferably S,and that the amino acid at position 301 is preferably A, and that the amino acid at position 17 is preferably P; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 79 is different from E and the amino acid at position 301 is different from D and the amino acid at position 40 is different from K, wherein it is preferred that the amino acid at position 186 is preferably W or Y, more preferably Y, and that the amino acid at position 79 is preferably S, W or I, more preferably S, and that the amino acid at position 301 is preferably A, and that the amino acid at position 40 is preferably M, wherein it is particularly preferred,that the amino acid at position 186 is Y and that the amino acid at position 79 is S and that the amino acid at position 301 is A and that the amino acid at position 40 is M; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 79 is different from E and the amino acid at position 301 is different from D and the amino acid at position 111 is different from Q, wherein it is preferred that the amino acid at position BCS221003-Ausland Dr. ML 23 186 is preferably W or Y, and that the amino acid at position 79 is preferably S, W or I, more preferably S, and that the amino acid at position 301 is preferably A, and that the amino acid at position 111 is preferably E; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 79 is different from E and the amino acid at position 3 is different from M and the amino acid at position 11 is different from C, wherein it is preferred,that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 79 is preferably S, W or I, more preferably S, and that the amino acid at position 3 is preferably Q, and that the amino acid at position 11 is preferably A; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 79 is different from E and the amino acid at position 3 is different from M and the amino acid at position 17 is different from L, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 79 is preferably S, W or I, more preferably S, and that the amino acid at position 3 is preferably Q,and that the amino acid at position 17 is preferably P; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 79 is different from E and the amino acid at position 3 is different from M and the amino acid at position 40 is different from K, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 79 is preferably S, W or I, more preferably S, and that the amino acid at position 3 is preferably Q, and that the amino acid at position 40 is preferably M; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 79 is different from E and the amino acid at position 3 is different from M and the amino acid at position 111 is different from Q, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 79 is preferably S, W or I, more preferably S,and that the amino acid at position 3 is preferably Q, and that the amino acid at position 111 is preferably E; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 79 is different from E and the amino acid at position 11 is different from C and the amino acid at position 17 is different from L, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 79 is preferably S, W or I, more preferably S, and that the amino acid at position 11 is preferably A, and that the amino acid at position 17 is preferably P; BCS221003-Ausland Dr. ML 24 ^ ^ the amino acid at position 186 is different from L and the amino acid at position 79 is different from E and the amino acid at position 11 is different from C and the amino acid at position 40 is different from K, wherein it is preferred that the amino acid at position 186 is preferably W or Y,and that the amino acid at position 79 is preferably S, W or I, more preferably S, and that the amino acid at position 11 is preferably A, and that the amino acid at position 40 is preferably M; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 79 is different from E and the amino acid at position 11 is different from C and the amino acid at position 111 is different from Q, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 79 is preferably S, W or I, more preferably S, and that the amino acid at position 11 is preferably A, and that the amino acid at position 111 is preferably E; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 79 is different from E and the amino acid at position 17 is different from L and the amino acid at position 40 is different from K, wherein it is preferred,that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 79 is preferably S, W or I, more preferably S, and that the amino acid at position 17 is preferably P, and that the amino acid at position 40 is preferably M; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 79 is different from E and the amino acid at position 17 is different from L and the amino acid at position 111 is different from Q, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 79 is preferably S, W or I, more preferably S, and that the amino acid at position 17 is preferably P,and that the amino acid at position 111 is preferably E; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 79 is different from E and the amino acid at position 40 is different from K and the amino acid at position 111 is different from Q, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 79 is preferably S, W or I, more preferably S, and that the amino acid at position 40 is preferably M, and that the amino acid at position 111 is preferably E; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 202 is different from A and the amino acid at position 280 is different from L and the amino acid at position 301 is different from D, wherein it is preferred that the amino acid BCS221003-Ausland Dr. ML 25 at position 186 is preferably W or Y, and that the amino acid at position 202 is preferably N,and that the amino acid at position 280 is preferably A, and that the amino acid at position 301 is preferably A; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 202 is different from A and the amino acid at position 280 is different from L and the amino acid at position 3 is different from M, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 202 is preferably N, and that the amino acid at position 280 is preferably A, and that the amino acid at position 3 is preferably Q; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 202 is different from A and the amino acid at position 280 is different from L and the amino acid at position 11 is different from C, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 202 is preferably N,and that the amino acid at position 280 is preferably A, and that the amino acid at position 11 is preferably A; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 202 is different from A and the amino acid at position 280 is different from L and the amino acid at position 17 is different from L, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 202 is preferably N, and that the amino acid at position 280 is preferably A, and that the amino acid at position 17 is preferably P; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 202 is different from A and the amino acid at position 280 is different from L and the amino acid at position 40 is different from K, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 202 is preferably N,and that the amino acid at position 280 is preferably A, and that the amino acid at position 40 is preferably M; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 202 is different from A and the amino acid at position 280 is different from L and the amino acid at position 111 is different from Q, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 202 is preferably N, and that the amino acid at position 280 is preferably A, and that the amino acid at position 111 is preferably E; BCS221003-Ausland Dr. ML 26 ^ ^ the amino acid at position 186 is different from L and the amino acid at position 202 is different from A and the amino acid at position 301 is different from D and the amino acid at position 3 is different from M, wherein it is preferred that the amino acid at position 186 is preferably W or Y,and that the amino acid at position 202 is preferably N, and that the amino acid at position 301 is preferably A, and that the amino acid at position 3 is preferably Q; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 202 is different from A and the amino acid at position 301 is different from D and the amino acid at position 11 is different from C, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 202 is preferably N, and that the amino acid at position 301 is preferably A, and that the amino acid at position 11 is preferably A; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 202 is different from A and the amino acid at position 301 is different from D and the amino acid at position 17 is different from L, wherein it is preferred that the amino acid at position 186 is preferably W or Y,and that the amino acid at position 202 is preferably N, and that the amino acid at position 301 is preferably A, and that the amino acid at position 17 is preferably P; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 202 is different from A and the amino acid at position 301 is different from D and the amino acid at position 40 is different from K, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 202 is preferably N, and that the amino acid at position 301 is preferably A, and that the amino acid at position 40 is preferably M; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 202 is different from A and the amino acid at position 301 is different from D and the amino acid at position 111 is different from Q, wherein it is preferred that the amino acid at position 186 is preferably W or Y,and that the amino acid at position 202 is preferably N, and that the amino acid at position 301 is preferably A, and that the amino acid at position 111 is preferably E; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 202 is different from A and the amino acid at position 3 is different from M and the amino acid at position 11 is different from C, wherein it is preferred that the amino acid at position BCS221003-Ausland Dr. ML 27 186 is preferably W or Y, and that the amino acid at position 202 is preferably N, and that the amino acid at position 3 is preferably Q, and that the amino acid at position 11 is preferably A; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 202 is different from A and the amino acid at position 3 is different from M and the amino acid at position 17 is different from L, wherein it is preferred,that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 202 is preferably N, and that the amino acid at position 3 is preferably Q, and that the amino acid at position 17 is preferably P; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 202 is different from A and the amino acid at position 3 is different from M and the amino acid at position 40 is different from K, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 202 is preferably N, and that the amino acid at position 3 is preferably Q, and that the amino acid at position 40 is preferably M; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 202 is different from A and the amino acid at position 3 is different from M and the amino acid at position 111 is different from Q, wherein it is preferred,that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 202 is preferably N, and that the amino acid at position 3 is preferably Q, and that the amino acid at position 111 is preferably E; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 202 is different from A and the amino acid at position 11 is different from C and the amino acid at position 17 is different from L, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 202 is preferably N, and that the amino acid at position 11 is preferably A, and that the amino acid at position 17 is preferably P; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 202 is different from A and the amino acid at position 11 is different from C and the amino acid at position 40 is different from K, wherein it is preferred,that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 202 is preferably N, and that the amino acid at position 11 is preferably A, and that the amino acid at position 40 is preferably M; BCS221003-Ausland Dr. ML 28 ^ ^ the amino acid at position 186 is different from L and the amino acid at position 202 is different from A and the amino acid at position 11 is different from C and the amino acid at position 111 is different from Q, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 202 is preferably N, and that the amino acid at position 11 is preferably A, and that the amino acid at position 111 is preferably E; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 202 is different from A and the amino acid at position 17 is different from L and the amino acid at position 40 is different from K,wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 202 is preferably N, and that the amino acid at position 17 is preferably P, and that the amino acid at position 40 is preferably M; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 202 is different from A and the amino acid at position 17 is different from L and the amino acid at position 111 is different from Q, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 202 is preferably N, and that the amino acid at position 17 is preferably P, and that the amino acid at position 111 is preferably E; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 202 is different from A and the amino acid at position 40 is different from K and the amino acid at position 111 is different from Q,wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 202 is preferably N, and that the amino acid at position 40 is preferably M, and that the amino acid at position 111 is preferably E; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 280 is different from L and the amino acid at position 301 is different from D and the amino acid at position 3 is different from M, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 280 is preferably A, and that the amino acid at position 301 is preferably A, and that the amino acid at position 3 is preferably Q; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 280 is different from L and the amino acid at position 301 is different from D and the amino acid at position 11 is different from C,wherein it is preferred that the amino acid BCS221003-Ausland Dr. ML 29 at position 186 is preferably W or Y, and that the amino acid at position 280 is preferably A, and that the amino acid at position 301 is preferably A, and that the amino acid at position 11 is preferably A; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 280 is different from L and the amino acid at position 301 is different from D and the amino acid at position 17 is different from L, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 280 is preferably A, and that the amino acid at position 301 is preferably A,and that the amino acid at position 17 is preferably P; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 280 is different from L and the amino acid at position 301 is different from D and the amino acid at position 40 is different from K, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 280 is preferably A, and that the amino acid at position 301 is preferably A, and that the amino acid at position 40 is preferably M; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 280 is different from L and the amino acid at position 301 is different from D and the amino acid at position 111 is different from Q, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 280 is preferably A, and that the amino acid at position 301 is preferably A,and that the amino acid at position 111 is preferably E; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 280 is different from L and the amino acid at position 3 is different from M and the amino acid at position 11 is different from C, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 280 is preferably A, and that the amino acid at position 3 is preferably Q, and that the amino acid at position 11 is preferably A; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 280 is different from L and the amino acid at position 3 is different from M and the amino acid at position 17 is different from L, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 280 is preferably A, and that the amino acid at position 3 is preferably Q,and that the amino acid at position 17 is preferably P; BCS221003-Ausland Dr. ML 30 ^ ^ the amino acid at position 186 is different from L and the amino acid at position 280 is different from L and the amino acid at position 3 is different from M and the amino acid at position 40 is different from K, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 280 is preferably A, and that the amino acid at position 3 is preferably Q, and that the amino acid at position 40 is preferably M; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 280 is different from L and the amino acid at position 3 is different from M and the amino acid at position 111 is different from Q, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 280 is preferably A,and that the amino acid at position 3 is preferably Q, and that the amino acid at position 111 is preferably E; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 280 is different from L and the amino acid at position 11 is different from C and the amino acid at position 17 is different from L, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 280 is preferably A, and that the amino acid at position 11 is preferably A, and that the amino acid at position 17 is preferably P; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 280 is different from L and the amino acid at position 11 is different from C and the amino acid at position 40 is different from K, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 280 is preferably A,and that the amino acid at position 11 is preferably A, and that the amino acid at position 40 is preferably M; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 280 is different from L and the amino acid at position 11 is different from C and the amino acid at position 111 is different from Q, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 280 is preferably A, and that the amino acid at position 11 is preferably A, and that the amino acid at position 111 is preferably E; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 280 is different from L and the amino acid at position 17 is different from L and the amino acid at position 40 is different from K, wherein it is preferred that the amino acid at position BCS221003-Ausland Dr. ML 31 186 is preferably W or Y,and that the amino acid at position 280 is preferably A, and that the amino acid at position 17 is preferably P, and that the amino acid at position 40 is preferably M; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 280 is different from L and the amino acid at position 17 is different from L and the amino acid at position 111 is different from Q, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 280 is preferably A, and that the amino acid at position 17 is preferably P, and that the amino acid at position 111 is preferably E; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 280 is different from L and the amino acid at position 40 is different from K and the amino acid at position 111 is different from Q, wherein it is preferred that the amino acid at position 186 is preferably W or Y,and that the amino acid at position 280 is preferably A, and that the amino acid at position 40 is preferably M, and that the amino acid at position 111 is preferably E; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 301 is different from D and the amino acid at position 3 is different from M and the amino acid at position 11 is different from C, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 301 is preferably A, and that the amino acid at position 3 is preferably Q, and that the amino acid at position 11 is preferably A; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 301 is different from D and the amino acid at position 3 is different from M and the amino acid at position 17 is different from L, wherein it is preferred that the amino acid at position 186 is preferably W or Y,and that the amino acid at position 301 is preferably A, and that the amino acid at position 3 is preferably Q, and that the amino acid at position 17 is preferably P; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 301 is different from D and the amino acid at position 3 is different from M and the amino acid at position 40 is different from K, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 301 is preferably A, and that the amino acid at position 3 is preferably Q, and that the amino acid at position 40 is preferably M; BCS221003-Ausland Dr. ML 32 ^ ^ the amino acid at position 186 is different from L and the amino acid at position 301 is different from D and the amino acid at position 3 is different from M and the amino acid at position 111 is different from Q, wherein it is preferred,that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 301 is preferably A, and that the amino acid at position 3 is preferably Q, and that the amino acid at position 111 is preferably E; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 301 is different from D and the amino acid at position 11 is different from C and the amino acid at position 17 is different from L, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 301 is preferably A, and that the amino acid at position 11 is preferably A, and that the amino acid at position 17 is preferably P; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 301 is different from D and the amino acid at position 11 is different from C and the amino acid at position 40 is different from K, wherein it is preferred,that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 301 is preferably A, and that the amino acid at position 11 is preferably A, and that the amino acid at position 40 is preferably M; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 301 is different from D and the amino acid at position 11 is different from C and the amino acid at position 111 is different from Q, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 301 is preferably A, and that the amino acid at position 11 is preferably A, and that the amino acid at position 111 is preferably E; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 301 is different from D and the amino acid at position 17 is different from L and the amino acid at position 40 is different from K, wherein it is preferred,that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 301 is preferably A, and that the amino acid at position 17 is preferably P, and that the amino acid at position 40 is preferably M; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 301 is different from D and the amino acid at position 17 is different from L and the amino acid at position 111 is different from Q, wherein it is preferred that the amino acid at position BCS221003-Ausland Dr. ML 33 186 is preferably W or Y, and that the amino acid at position 301 is preferably A, and that the amino acid at position 17 is preferably P, and that the amino acid at position 111 is preferably E; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 301 is different from D and the amino acid at position 40 is different from K and the amino acid at position 111 is different from Q,wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 301 is preferably A, and that the amino acid at position 40 is preferably M, and that the amino acid at position 111 is preferably E; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 3 is different from M and the amino acid at position 11 is different from C and the amino acid at position 17 is different from L, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 3 is preferably Q, and that the amino acid at position 11 is preferably A, and that the amino acid at position 17 is preferably P; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 3 is different from M and the amino acid at position 11 is different from C and the amino acid at position 40 is different from K, wherein it is preferred,that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 3 is preferably Q, and that the amino acid at position 11 is preferably A, and that the amino acid at position 40 is preferably M; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 3 is different from M and the amino acid at position 11 is different from C and the amino acid at position 111 is different from Q, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 3 is preferably Q, and that the amino acid at position 11 is preferably A, and that the amino acid at position 111 is preferably E; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 3 is different from M and the amino acid at position 17 is different from L and the amino acid at position 40 is different from K, wherein it is preferred,that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 3 is preferably Q, and that the amino acid at position 17 is preferably P, and that the amino acid at position 40 is preferably M; BCS221003-Ausland Dr. ML 34 ^ ^ the amino acid at position 186 is different from L and the amino acid at position 3 is different from M and the amino acid at position 17 is different from L and the amino acid at position 111 is different from Q, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 3 is preferably Q, and that the amino acid at position 17 is preferably P, and that the amino acid at position 111 is preferably E; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 3 is different from M and the amino acid at position 40 is different from K and the amino acid at position 111 is different from Q,wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 3 is preferably Q, and that the amino acid at position 40 is preferably M, and that the amino acid at position 111 is preferably E; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 11 is different from C and the amino acid at position 17 is different from L and the amino acid at position 40 is different from K, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 11 is preferably A, and that the amino acid at position 17 is preferably P, and that the amino acid at position 40 is preferably M; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 11 is different from C and the amino acid at position 17 is different from L and the amino acid at position 111 is different from Q, wherein it is preferred,that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 11 is preferably A, and that the amino acid at position 17 is preferably P, and that the amino acid at position 111 is preferably E; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 11 is different from C and the amino acid at position 40 is different from K and the amino acid at position 111 is different from Q, wherein it is preferred that the amino acid at position 186 is preferably W or Y, and that the amino acid at position 11 is preferably A, and that the amino acid at position 40 is preferably M, and that the amino acid at position 111 is preferably E; ^ ^ the amino acid at position 186 is different from L and the amino acid at position 17 is different from L and the amino acid at position 40 is different from K and the amino acid at position 111 is different from Q, wherein it is preferred,that the amino acid at position BCS221003-Ausland Dr. ML 35 186 is preferably W or Y, and that the amino acid at position 17 is preferably P, and that the amino acid at position 40 is preferably M, and that the amino acid at position 111 is preferably E; ^ ^ Proteins with an amino acid sequence which is at least 80%, preferably 85%, more preferably 90%, even more preferably 95%, even more preferably 96%, even more preferably 97%, particularly preferably 98%, most preferably 99% identical to the amino acid sequence shown under a), provided that the amino acid at position 186 is different from L, wherein it is preferred that the amino acid at position 186 is preferably W or Y, more preferably Y, and wherein the proteins have at least three further amino acid substitutions selected from the group,which is shown under the listed symbols directly above. The lipase variants described above under point a) with respect to the amino acid sequences shown under SEQ ID No. 1 may have at least four further amino acid substitutions at positions 79, 202, 280, 301, 3, 11, 17, 40 or 111. Preferably, the amino acid at position 79 is S, W or I,more preferably S; preferably the amino acid at position 202 is N; preferably the amino acid at position 280 is A; preferably the amino acid at position 301 is A; preferably the amino acid at position 3 is Q; preferably the amino acid at position 11 is A; preferably the amino acid at position 17 is P; preferably the amino acid at position 40 is M; preferably the amino acid at position 111 is E. The lipase variants described herein above under point a) with respect to the amino acid sequences shown under SEQ ID No. 1 may have at least five further amino acid substitutions at positions 79, 202, 280, 301, 3, 11, 17, 40 or 111. Preferably, the amino acid at position 79 is S, W or I,more preferably S; preferably the amino acid at position 202 is N; preferably the amino acid at position 280 is A; preferably the amino acid at position 301 is A; preferably the amino acid at position 3 is Q; preferably the amino acid at position 11 is A; preferably the amino acid at position 17 is P; preferably the amino acid at position 40 is M; preferably the amino acid at position 111 is E. The lipase variants described herein above under point a) with respect to the amino acid sequences shown under SEQ ID No. 1 may have at least six further amino acid substitutions at positions 79, 202, 280, 301, 3, 11, 17, 40 or 111. Preferably the amino acid at position 79 is S, W or I,more preferably S; preferably the amino acid at position 202 is N; preferably the amino acid at position 280 is A; preferably the amino acid at position 301 is A; preferably the amino acid at position 3 is Q; preferably the amino acid at position 11 is A; BCS221003-Ausland Dr. ML 36 preferably the amino acid at position 17 is P; preferably the amino acid at position 40 is M; preferably the amino acid at position 111 is E. The lipase variants described herein above under point a) with respect to the amino acid sequences shown under SEQ ID No. 1 can have at least seven further amino acid substitutions at positions 79, 202, 280, 301, 3, 11, 17, 40 or 111. Preferably the amino acid at position 79 is S, W or I,more preferably S; preferably the amino acid at position 202 is N; preferably the amino acid at position 280 is A; preferably the amino acid at position 301 is A; preferably the amino acid at position 3 is Q; preferably the amino acid at position 11 is A; preferably the amino acid at position 17 is P; preferably the amino acid at position 40 is M; Preferably, the amino acid at position 111 is E. Preferred embodiments according to the invention are proteins which are encoded for lipases having the amino acids listed under SEQ ID Nos. 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149, 151, 153, 155, 157, 159, 161, 163, 165, 167, 169, 171, 173, 175, 177, 179, 181, 183, 185, 187, 189, 191, 193, 195, 197, 199, 201, 203, 205, 207, 209, 211,213, 215, 217, 219, 221, 223, 225, 227, 229, 231, 233, 235, 237, 239, 241, 243, 245, 247, 249, 251, 253, 255, 257, 259, 261, 263, 265, 267, 269, 271, 273, 275, 277, 279, 281, 283, 285, 287, 289, 291, 293, 295, 297, 299, 301, 303, 305, 307, 309, 311, 313, 315, 317, 319, 321, 323, 325, 327, 329, 331, 333, 335, 337, 339, 341, 343, 345, 347, 349, 351, 353, 355, 357, 359, 361, 363, 365, 367, 369, 371, 373, 375, 377, 379, 381, 383, 385, 387, 389, 391, 393, 395, 397, 399, 401. Particularly preferred embodiments of the invention are proteins encoding lipases with the amino acid sequences shown under SEQ ID Nos. 233 and 399. Further proteins with the activity of a lipolytic enzyme or a lipase have been tested. The amino acid sequences of these further proteins represent variants of the amino acid sequence represented by the amino acid in SEQ ID No. 1.where: ^ ^ in the amino acid sequence shown under SEQ ID No. 1, the two amino acids at positions 40 and 79 are different from the amino acids indicated at the corresponding amino acid positions in the sequence shown under SEQ ID No. 1. In a specific tested variant with respect to the amino acid sequence shown under SEQ ID No. 1, the amino acid at position 40 is M and the amino acid at position 79 is S. ^ ^ in the amino acid sequence shown under SEQ ID No. 1, the two amino acids at positions 40 and 186 are different from the amino acids indicated at the corresponding amino acid positions BCS221003-Ausland Dr. ML 37 in the sequence shown under SEQ ID No. 1.different. In a specific tested variant with respect to the amino acid sequence shown under SEQ ID No. 1, the amino acid at position 40 is M and the amino acid at position 186 is Y; ^ ^ in the amino acid sequence shown under SEQ ID No. 1, the two amino acids at positions 40 and 301 are different from the amino acids indicated at the corresponding amino acid positions in the sequence shown under SEQ ID No. 1. In a specific tested variant with respect to the amino acid sequence shown under SEQ ID No. 1, the amino acid at position 40 is M and the amino acid at position 301 is A; ^ ^ in the amino acid sequence shown under SEQ ID No. 1, the two amino acids at positions 79 and 186 are different from the amino acids indicated at the corresponding amino acid positions in the sequence shown under SEQ ID No. 1.different. In a specific tested variant with respect to the amino acid sequence shown under SEQ ID No. 1, the amino acid at position 79 is S and the amino acid at position 186 is Y; ^ ^ in the amino acid sequence shown under SEQ ID No. 1, the two amino acids at positions 79 and 301 are different from the amino acids indicated at the corresponding amino acid positions in the sequence shown under SEQ ID No. 1. In a specific tested variant with respect to the amino acid sequence shown under SEQ ID No. 1, the amino acid at position 79 is S and the amino acid at position 301 is A; ^ ^ in the amino acid sequence shown under SEQ ID No. 1, the two amino acids at positions 186 and 301 are different from the amino acids indicated at the corresponding amino acid positions in the sequence shown under SEQ ID No. 1.different. In a specific tested variant with respect to the amino acid sequence shown under SEQ ID No. 1, the amino acid at position 186 is Y and the amino acid at position 301 is A. Further proteins with the activity of a lipolytic enzyme or a lipase have also been tested. The amino acid sequences of these further proteins represent variants of the amino acid sequence shown by the amino acid in SEQ ID No. 1, where: ^ ^ in the amino acid sequence shown under SEQ ID No. 1, the three amino acids at positions 40, 79 and 186 are different from the amino acids indicated at the corresponding amino acid positions in the sequence shown under SEQ ID No. 1,different. In a particular variant tested with respect to the amino acid sequence shown under SEQ ID No. 1, the amino acid at position 40 is M and the amino acid at position 79 is S and the amino acid at position 186 is Y; BCS221003-Ausland Dr. ML 38 ^ ^ in the amino acid sequence shown under SEQ ID No. 1, the three amino acids at positions 40, 79 and 301 are different from the amino acids indicated at the corresponding amino acid positions in the sequence shown under SEQ ID No. 1. In a particular variant tested with respect to the amino acid sequence shown under SEQ ID No. 1, the amino acid at position 40 is M and the amino acid at position 79 is S and the amino acid at position 301 is A; ^ ^ in the amino acid sequence shown under SEQ ID No.1, the three amino acids at positions 40, 186 and 301 are different from the amino acids indicated at the corresponding amino acid positions in the sequence shown under SEQ ID No.1,different. In a specific tested variant with respect to the amino acid sequence shown under SEQ ID No. 1, the amino acid at position 40 is M and the amino acid at position 186 is Y and the amino acid at position 301 is A; ^ ^ in the amino acid sequence shown under SEQ ID No. 1, the three amino acids at positions 79, 186 and 301 are different from the amino acids indicated at the corresponding amino acid positions in the sequence shown under SEQ ID No. 1. In a specific tested variant with respect to the amino acid sequence shown under SEQ ID No. 1, the amino acid at position 79 is S and the amino acid at position 186 is Y and the amino acid at position 301 is A; The lipases and lipase variants according to the invention have a high selectivity and / or a high specific activity with respect to the stereoselective acylation or carboxylation of 2,6-dimethyl-1-indanamine (DMAI) and are capable ofto produce enantiomerically enriched or nearly pure methyl [(1R,2S)-2,6-dimethyl-2,3-dihydro-1H-inden-1-yl] carbamate. Methyl [(1R,2S)-2,6-dimethyl-2,3-dihydro-1H-inden-1-yl] carbamate is an important intermediate for the synthesis of the herbicidal compound indaziflam. “Enantiomerically enriched” means herein that one of two enantiomers is present in a composition in greater amounts than the other enantiomer, preferably one enantiomer is present in the composition to at least 60%, more preferably one enantiomer is present in the composition to at least 65%, even more preferably one enantiomer is present in the composition to at least 70%, even more preferably one enantiomer is present in the composition to at least 75%, even more preferably one enantiomer is present in the composition to at least 80%,Particularly preferably, one enantiomer is present in the composition to at least 85%, most preferably, one enantiomer is present in the composition to at least 90%, or very particularly preferably, one enantiomer is present in the composition to at least 94%. "Enantiomerically nearly pure" means herein that one of two enantiomers is present in a composition in amounts of at least 95.0%, preferably one of two enantiomers is present in a composition in amounts of at least 95.5%, more preferably one of two enantiomers is present in a composition in amounts of at least 96.0%, even more preferably one of two enantiomers is present in a composition in amounts of at least 96.5%, even more preferably one of two enantiomers is present in a composition in amounts of at least 97.0%,Even more preferably, one of two enantiomers is present in a composition in amounts of at least 98.0%, particularly preferably one of two enantiomers is present in a composition in amounts of at least 98.5%, most preferably one of two enantiomers is present in a composition in amounts of at least 99.0%, or most preferably one of two enantiomers is present in a composition in amounts of at least 99.5%. A further embodiment of the invention relates to nucleic acid molecules that encode a protein according to the invention. Nucleic acid molecules according to the invention can be any type of nucleic acid, provided that the nucleic acid encodes a protein according to the invention. The nucleic acids can be ribonucleic acid molecules (e.g., RNA, mRNA) or deoxyribonucleic acid molecules (DNA, including genomic DNA,which may or may not include introns and coding DNA). Of particular interest for the invention are nucleic acid molecules which encode proteins which have the activity of a lipase, comprising those listed under SEQ ID Nos. 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149, 151, 153, 155, 157, 159, 161, 163, 165, 167, 169, 171, 173, 175, 177, 179, 181, 183, 185, 187, 189, 191, 193, 195, 197, 199, 201, 203, 205, 207, 209, 211, 213, 215, 217, 219, 221, 223, 225, 227, 229, 231, 233, 235, 237, 239, 241, 243, 245, 247, 249, 251, 253, 255, 257, 259, 261, 263, 265, 267, 269, 271, 273, 275, 277, 279, 281, 283, 285, 287, 289, 291, 293, 295, 297, 299, 301, 303, 305, 307, 309, 311, 313, 315, 317, 319, 321, 323,325, 327, 329, 331, 333, 335, 337, 339, 341, 343, 345, 347, 349, 351, 353, 355, 357, 359, 361, 363, 365, 367, 369, 371, 373, 375, 377, 379, 381, 383, 385, 387, 389, 391, 393, 395, 397, 399, 401. BCS221003-Ausland Dr. ML 40 The invention further relates to nucleic acid molecules which code for a protein with the activity of a lipase, selected from the group consisting of a) nucleic acid molecules which have the sequences listed under SEQ ID Nos. 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150, 152, 154, 156, 158, 160, 162, 164, 166, 168, 170, 172, 174, 176, 178, 180, 182, 184, 186, 188, 190, 192, 194, 196, 198, 200, 202, 204, 206, 208, 210, 212, 214, 216, 218, 220, 222, 224, 226, 228, 230, 232, 234,236, 238, 240, 242, 244, 246, 248, 250, 252, 254, 256, 258, 260, 262, 264, 266, 268, 270, 272, 274, 276, 278, 280, 282, 284, 286, 288, 290, 292, 294, 296, 298, 300, 302, 304, 306, 308, 310, 312, 314, 316, 318, 320, 322, 324, 326, 328, 330, 332, 334, 336, 338, 340, 342, 344, 346, 348, 350, 352, 354, 356, 358, 360, 362, 364, 366, 368, 370, 372, 374, 376, 378, 380, 382, ​​384, 386, 388, 390, 392, 394, 396, 398, 400, 402; b) nucleic acid molecules which have at least 60%, preferably 70%, more preferably 80%, even more preferably 90%, even more preferably 95%, even more preferably 96%, particularly preferably 97%, most preferably 98% or most preferably 99% identity to the nucleic acid sequences shown under a). In the context of the present invention, "hybridize with" means hybridization under conventional hybridization conditions, preferably under stringent conditions,as described, for example, in Sambrook et al. (Molecular Cloning, A Laboratory Manual, 3rd edition (2001) Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY. ISBN: 0879695773) or in Ausubel et al. (Short Protocols in Molecular Biology, John Wiley & Sons; 5th edition (2002), ISBN: 0471250929). Particularly preferably, “hybridization” means hybridization under the following conditions: hybridization buffer: 2xSSC; 10xDenhardt’s solution (Fikoll 400+PEG+BSA; ratio 1:1:1); 0.1% SDS; 5 mM EDTA; 50 mM Na2HPO4; 250 µg / ml herring sperm DNA; 50 µg / ml tRNA; or 25 M sodium phosphate buffer, pH 7.2; 1 mM EDTA; 7% SDS Hybridization temperature: T = 65 to 68 °C Wash buffer: 0.1xSSC; 0.1% SDS Wash temperature: T = 65 to 68 °C. BCS221003-Ausland Dr. ML 41 Nucleic acid molecules that hybridize with nucleic acid molecules encoding a protein with lipase activity can originate from any organism; accordingly, they can be derived from bacteria, fungi,Animals, humans, plants, or viruses. Nucleic acid molecules that hybridize with nucleic acid molecules encoding a protein with lipase activity preferably originate from microorganisms, more preferably from fungi or bacteria, most preferably from bacteria. Nucleic acid molecules that hybridize with said molecules can be isolated, for example, from genomic or cDNA libraries. These nucleic acid molecules can be identified and isolated using the nucleic acid molecules described herein, or they can be identified and isolated using parts of these molecules or the reverse complements of these molecules, for example, by hybridization according to standard procedures (see, for example, Sambrook et al., Molecular Cloning, A Laboratory Manual, 3rd ed. (2001) Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY. ISBN: 0879695773; Ausubel et al.,Short Protocols in Molecular Biology, John Wiley & Sons; 5th edition (2002), ISBN: 0471250929) or by amplification using PCR. The fragments used as hybridization probes can also be synthetic fragments or oligonucleotides prepared using conventional synthesis techniques, the sequence of which is essentially identical to the nucleic acid molecule described in the context of the present invention. If genes that hybridize with the nucleic acid sequences described in the context of the present invention are identified and isolated, the sequence should be determined and the properties of the proteins encoded by this sequence should be analyzed to determine whether they are proteins with lipase activity. Method for determining whether a protein has lipase activityare known to those of ordinary skill in the art. The molecules that hybridize with the nucleic acid molecules described in the present invention include, in particular, fragments, derivatives, and allelic variants of said nucleic acid molecules. In the context of the present invention, the term "derivative" means that the sequences of these molecules differ in one or more positions from the sequences of the nucleic acid molecules described above and are highly identical to these sequences. The differences from the nucleic acid molecules described above can be attributed, for example, to deletion, addition, substitution, insertion, or recombination. Preferred nucleic acid molecules according to the invention are those under SEQ ID Nos. 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108,110, 112, 114, 116, 118, BCS221003-Abroad Dr. ML 42 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150, 152, 154, 156, 158, 160, 162, 164, 166, 168, 170, 172, 174, 176, 178, 180, 182, 184, 186, 188, 190, 192, 194, 196, 198, 200, 202, 204, 206, 208, 210, 212, 214, 216, 218, 220, 222, 224, 226, 228, 230, 232, 234, 236, 238, 240, 242, 244, 246, 248, 250, 252, 254, 256, 258, 260, 262, 264, 266, 268, 270, 272, 274, 276, 278, 280, 282, 284, 286, 288, 290, 292, 294, 296, 298, 300, 302, 304, 306, 308, 310, 312, 314, 316, 318, 320, 322, 324, 326, 328, 330, 332, 334, 336, 338, 340, 342, 344, 346, 348, 350, 352, 354, 356, 358, 360, 362, 364, 366, 368, 370, 372, 374, 376, 378, 380, 382, ​​384, 386, 388, 390, 392, 394, 396, 398, 400, 402. The meaning of the nucleotide abbreviations a, c, g, t as well as the abbreviations for degenerate nucleotides r, y, s, w, k, m, b, d, h, v,n can be derived hereinafter from Table 1 in the section entitled "Description of the Sequences". The amino acids encoded by degenerate nucleotides can be derived hereinafter from Table 3 in the section entitled "Description of the Sequences". Also disclosed are recombinant nucleic acid molecules which comprise a nucleic acid molecule suitable for the method according to the invention. The term "recombinant nucleic acid molecule" is understood to mean a nucleic acid molecule which, in addition to the nucleic acid molecule suitable for the method according to the invention, contains further sequences which do not occur naturally in the combination in which they occur in the nucleic acids recombinant for the method according to the invention. The above-mentioned additional sequences can be any sequences, preferably they are functional or regulatory sequences (promoters, termination signals,Enhancers, ribosome binding sites (rbs), leader sequences that increase transcription, translation, or RNA stability, subcellular targeting sequences, etc.), particularly preferably functional or regulatory sequences that are active in microorganisms, and very particularly preferably regulatory sequences that are active in fungi, in particular yeasts, or bacteria. Methods for producing the recombinant nucleic acid molecules suitable for the method according to the invention are known to those of ordinary skill in the art. These include genetic methods, such as binding nucleic acid molecules by ligation, genetic recombination, or de novo synthesis of nucleic acid molecules. These methods are described, for example, in Sambrok et al. (Molecular Cloning, A Laboratory Manual, 3rd edition (2001) Cold Spring Harbour Laboratory Press, Cold Spring Harbour, NY. ISBN: 0879695773) or in Ausubel et al. (Short Protocols in Molecular Biology,John Wiley & Sons; 5th edition (2002), ISBN: 0471250929). BCS221003-Ausland Dr. ML 43 The recombinant nucleic acid molecule usable for the method according to the invention comprises a suitable nucleic acid molecule linked to regulatory sequences that initiate transcription in prokaryotic or eukaryotic cells. Regulatory sequences that initiate transcription in a cell are also known as promoters. Information relating to regulatory sequences and plasmids is well known to those of ordinary skill in the art and is described, for example, in the Registry of Standard Biological Parts, supported by The International Genetically Engineered Machine (iGEM) Foundation (One Kendall Square, Suite B6104, Cambridge, MA 02139, USA) on the Internet (http: / / parts.igem.org / Catalog). Regulatory sequences that initiate transcription in prokaryotic organisms, such as E. coli, as well as in eukaryotic organisms,are extensively described in the literature, particularly those relating to expression in yeasts, e.g., Saccharomyces cerevisiae. An overview of various systems for the expression of proteins in different host organisms can be found, for example, in Methods in Enzymology 153 (1987), 383–516, as well as in Bitter et al. (Methods in Enzymology 153 (1987), 516–544), or in Gomes et al. (2016, Advances in Animal and Veterinary Sciences, 4(4), 346) and Baghban et al. (2018, Current Pharmaceutical Biotechnology, 19(6)). Common yeast promoters are pAOX1, pHIS4, pGAL, and pScADH2 (Baghban et al., 2018, see above). Common bacterial promoters are T5, T7, rhamnose-inducible, arabinose-inducible, PhoA, artificial trc(trp-lac) promoter, as described by Marschall et al. (2017, Appl Microbiol Biotechnol 101, 501–512) and Tegel et al. (2011, FEBS Journal 278,729-739). Another embodiment of recombinant nucleic acid molecules usable for the method according to the invention are vectors of plasmids comprising the appropriate nucleic acid molecules. "Vectors" are well known in the field of molecular biology and herein represent a nucleic acid sequence or a vehicle comprising a nucleic acid sequence used to transfer genetic material (DNA or RNA) into a target cell. Vectors can be plasmids, e.g., T-DNA or binary vectors for generating transgenic plants, expression vectors for expressing nucleic acid sequences in a host cell, shuttle vectors capable of replicating in different hosts, or vectors can be virus particles or bacteriophages that have been modified.to deliver foreign genetic material into a host. "Plasmids" are well known in the field of molecular biology and, as used herein, represent an autonomously self-replicating, often circular DNA molecule which, when present in a host cell, is separated from the chromosomal DNA. BCS221003-Ausland Dr. ML 44 Suitable nucleic acid molecules, recombinant nucleic acid molecules, vectors, or plasmids can be used to produce proteins for the method according to the invention, e.g., by expressing the appropriate nucleic acid molecule in host cells. Also disclosed are hosts or host cells,which comprise or express a nucleic acid molecule suitable for the method according to the invention, or which comprise suitable proteins with the activity of a lipase, or which comprise a recombinant nucleic acid molecule suitable for the method according to the invention, or which comprise a vector suitable for the method according to the invention, or which comprise a plasmid suitable for the method according to the invention. Suitable nucleic acid molecules encoding a protein with the activity of a lipase can be expressed in host cells, e.g., for their replication or for the production of proteins with the activity of a lipase. For expression in host cells, suitable nucleic acid molecules can be contained on vectors or plasmids, or can be stably integrated into the genome of a respective host cell. The suitable nucleic acid molecules can also be contained in vectors,which support their introduction into host cells. Also disclosed is a host or host cell suitable for the method according to the invention, comprising a nucleic acid molecule suitable for the method according to the invention or comprising a recombinant nucleic acid molecule suitable for the method according to the invention or comprising a vector suitable for the method according to the invention or comprising a plasmid suitable for the method according to the invention and each comprising a protein suitable for the method according to the invention. Also disclosed is a host or host cell suitable for the method according to the invention,comprising a nucleic acid molecule suitable for the method according to the invention, or comprising a recombinant nucleic acid molecule suitable for the method according to the invention, or comprising a vector suitable for the method according to the invention, or comprising a plasmid suitable for the method according to the invention and each expressing a protein suitable for the method according to the invention, wherein the protein preferably has the activity of a lipase. "Expressing a nucleic acid molecule" is to be understood herein as meaning that, if the nucleic acid molecule is RNA or mRNA, the nucleic acid molecule is translated into a protein, preferably into a protein with the activity of a lipase, or if the nucleic acid molecule is DNA or cDNA, it is transcribed into mRNA (and processed in the case of intron-containing genomic DNA), preferably into an mRNA encoding a protein with the activity of a lipase.and is subsequently translated into a protein, preferably into a protein with the activity of a lipase. BCS221003-Ausland Dr. ML 45 The transcription of a particular nucleic acid molecule in a host can be detected by methods known to those of ordinary skill in the art, for example, by detecting specific transcripts (mRNA) of foreign nucleic acid molecules by Northern blot analysis or by RT-PCR. Whether hosts or host cells comprise a particular protein or comprise a protein derived from the expression of a nucleic acid molecule can be determined by methods known to those of ordinary skill in the art, for example, by immunological methods such as Western blot analysis, ELISA (Enzyme-Linked ImmunoSorbent Assay), or RIA (Radio Immune Assay). Those of ordinary skill in the art are familiar with methods for producing antibodies that specifically react with a particular protein.i.e., which bind specifically to a particular protein (see, for example, Lottspeich and Zorbas (eds.), 1998, Bioanalytik, Spektrum akad, Verlag, Heidelberg, Berlin, ISBN 3-8274-0041-4). Some companies (Thermo Fisher Scientific, 168 Third Avenue, Waltham, MA USA 0245; GenScript, 60 Centennial Ave., Piscataway, NJ 08854, USA) offer the production of these antibodies as a service to order. Furthermore, those of ordinary skill in the art can check whether a host or host cell comprises a protein suitable for the method of the invention by detecting the (additional) activity of proteins with the activity of a lipase in a corresponding host cell. Preferably, the activity of proteins with additional activity of a lipase in a corresponding host cell is detected by comparing the activities of lipases in a host cell used for the method according to the invention with the corresponding activity of host cells,which do not comprise a protein suitable for the method according to the invention. Testing whether a protein exhibits lipase activity can be carried out using methods known in the art. Hosts or host cells used for the method according to the invention can be produced by those of ordinary skill in the art using known methods for genetically modifying or transforming organisms. Furthermore, a host or host cell suitable for the method according to the invention, in particular a prokaryotic or eukaryotic host or host cell, is disclosed.genetically modified (or transformed) with a suitable nucleic acid molecule or with a suitable recombinant nucleic acid molecule or with a suitable vector or with a suitable plasmid. Preferably, the genetically modified (transformed) host or host cell used for the method according to the invention expresses a protein with the activity of a lipase; more preferably, the genetically modified (transformed) host or host cell expresses a protein suitable for the method according to the invention. "Genetically modified with a nucleic acid molecule BCS221003-Ausland Dr. ML 46" or "transformed with a nucleic acid molecule" is to be understood herein to mean that a nucleic acid molecule is or has been introduced into a host or host cell by technical and / or non-naturally occurring means, preferably by technical methods from the field of molecular biology.biotechnology or genetic engineering. Descendants or progeny of hosts or host cells used for the method according to the invention are also disclosed; preferably, these descendants or progeny comprise a nucleic acid molecule suitable for the method according to the invention or comprise a suitable recombinant nucleic acid molecule or comprise a suitable vector or comprise a suitable plasmid or comprise a suitable protein; more preferably, these descendants or progeny comprise a suitable nucleic acid molecule or comprise a suitable recombinant nucleic acid molecule or comprise a suitable vector or comprise a suitable plasmid and in each case they express a protein, wherein the protein has the activity of a lipase.Even more preferably, these progeny or descendants comprise a suitable nucleic acid molecule or comprise a suitable recombinant nucleic acid molecule or comprise a suitable vector or comprise a suitable plasmid and in each case they express a protein, wherein the protein has the activity of a lipase that can be used in the process according to the invention. The host or host cell for the process according to the invention can be a host or host cell of a prokaryotic or a eukaryotic organism. The host or host cells can be bacteria or bacterial cells (e.g. E. coli, bacteria of the genus Bacillus, in particular Bacillus subtilis, Agrobacterium, in particular Agrobacterium tumefaciens or Agrobacterium rhizogenes, Pseudomonas, in particular Pseudomonas fluorescens, Streptomyces spp, Rhodococcus spp, in particular Rhodococcus rhodochrous, Vibrio natrigens, Corynebacterium,in particular Corynebacterium glutamicum) or fungi or fungal cells (e.g. Agaricus, in particular Agaricus bisporus, Aspergillus, Trichoderma or yeasts, in particular S. cerevisiae, Pichia ssp. such as P. pastoris), as well as plants or plant cells, or they can be animals or animal cells. Preferred host cells are cells of microorganisms. In the context of the present patent application, it is assumed that all bacteria and protists (e.g. fungi, in particular yeasts and algae), as defined, for example, in Schlegel "General Microbiology" (Georg Thieme Verlag (1985), 1-2), are included. With regard to microorganisms, the hosts or host cells used for the process according to the invention are preferably bacteria / bacterial cells or yeasts / yeast cells, more preferably bacteria / bacterial cells, even more preferably Bacillus species / Bacillus species cells or Escherichia coli / Escherichia coli cells,most preferably Escherichia coli / Escherichia coli cells. Alternatively, Pseudomonas, in particular Pseudomonas fluorescens, Streptomyces spp, Rhodococcus spp, in particular Rhodococcus rhodochrous, Vibrio spp, in particular Vibrio natrigens, Corynebacterium, in particular Corynebacterium glutamicum, or other hosts or host cells can be used for the method according to the invention. Preferred hosts or host cells for the method according to the invention comprise a suitable nucleic acid molecule, wherein the suitable nucleic acid molecule is characterized in that the codons of the nucleic acid molecule are modified such that they are adapted to the codon usage frequency of the host or a host cell. Description of the sequences Throughout the application, abbreviations for nucleotides and amino acids are used according to the following IUPAC codes: Table 1, BCS221003-Ausland Dr. ML 48 To distinguish between amino acids and nucleotides, the abbreviated nucleotide codes written in capital letters in the table above are written in lowercase here. Table 2 The use of codons herein follows the so-called "general genetic code" as shown in the following table, where "t" in ribonucleotide acid (RNA) sequences is to be replaced by "u." BCS221003-Ausland Dr. ML 49 Table 3 BCS221003-Abroad Dr. ML 50 BCS221003-Ausland Dr. ML 51 Table 4 The sequence listing associated with this application is submitted in electronic format and is hereby incorporated by reference in its entirety into this patent specification. "PRT" stands for "protein" and "NUC" for "nucleic acid." BCS221003-Abroad Dr. ML 52 BCS221003-Abroad Dr. ML 53 BCS221003-Abroad Dr. ML 54 BCS221003-Abroad Dr. ML 55 BCS221003-Abroad Dr. ML 56 BCS221003-Abroad Dr. ML 57 BCS221003-Abroad Dr. ML 58 BCS221003-Abroad Dr. ML 59 BCS221003-Abroad Dr. ML 60 BCS221003-Abroad Dr. ML 61 BCS221003-Abroad Dr. ML 62 BCS221003-Abroad Dr. ML 63 BCS221003-Abroad Dr. ML 64 BCS221003-Abroad Dr. ML 65 BCS221003-Abroad Dr. ML 66 The protein with lipase activity is preferably used in an amount of 0.1-50 wt.% based on the mixture (I). Particular preference is given to an amount of 0.5-10 wt.%. In particular preference is given to an amount of 1-5 wt.%. Step 1 of the process according to the invention can be carried out with or without a solvent. Preference is given to solvents from the group consisting of methyl t-butyl ether, heptane, toluene, xylenes, mesitylene, anisole, chlorobenzene, n-butanol, i-propanol, n-propanol and ethanol, and mixtures thereof. Particular preference is given to solvents from the group consisting of toluene, xylenes, mesitylene, n-butanol and ethanol, and mixtures thereof. Likewise, the reaction is particularly preferably carried out in the absence of a solvent. Based on the amount of mixture (I) used, the acylating or carboxylating agent R C(=O) R 1preferably used in an amount of 1-25 equivalents. More preferably it is used in an amount of 1-10 equivalents. In particular preferably it is used in an amount of 1-5 equivalents. The reaction according to step 1 is usually carried out at 0 to 10 bar hydrogen pressure. A hydrogen pressure of 1 to 5 bar is preferred. The reaction according to step 1 is usually carried out at temperatures of 40-130 °C. The reaction is preferably carried out at 70-130 °C. More preferably the reaction is carried out at 105-125 °C. BCS221003-Ausland Dr. ML 67 The metal catalyst used in step 1 is preferably a palladium on carbon (Pd / C) or a palladium on aluminum oxide (Pd / Al2O3) catalyst with a palladium loading of 0.5-10 wt. %.Also preferred is the Shvo catalyst with the IUPAC name 1-hydroxytetraphenylcyclopentadienyl-(tetraphenyl-2,4-cyclopentadien-1-one)-μ-hydrotetracarbonyldiruthenium(II) in a stoichiometry of 1-10 mol%. Particular preference is given to using a palladium-on-carbon (Pd / C) or a palladium-on-aluminum (Pd / Al2O3) catalyst with a palladium loading of 0.5-10 wt%. Particular preference is given to using a palladium-on-aluminum (Pd / Al2O3) catalyst with a palladium loading of 0.5-10 wt%. The catalysts are used in an amount of 0.1-10 wt% based on the compounds of formula (I); preferably, 0.5-5 wt% is used. The amount of catalysts used is calculated based on the dry mass of the catalysts.The separation of component (II) mentioned in step 2 of the process according to the invention is carried out by crystallization or another suitable purification process known to those skilled in the art. The bases mentioned in step 3 of the process according to the invention are typically selected from the group consisting of lithium hydroxide, sodium hydroxide, potassium hydroxide, lithium carbonate, sodium carbonate, potassium carbonate, lithium methoxide, sodium methoxide, potassium methoxide, lithium ethoxide, sodium ethoxide, and potassium ethoxide. Particular preference is given to using the bases lithium hydroxide, sodium hydroxide, potassium hydroxide, lithium ethoxide, sodium ethoxide, and potassium ethoxide. The bases lithium hydroxide, sodium hydroxide, potassium hydroxide, lithium ethoxide, sodium ethoxide, and potassium ethoxide are preferred. The base is typically used in a stoichiometry of 1.00-3.00 equivalents, based on the amount of component (II) used.The base is preferably used in an amount of 1.50-2.50 equivalents. The base is particularly preferably used in an amount of 1.75 to 2.25 equivalents. Preferred solvents are ethanol, n-propanol, i-propanol, n-butanol, i-butanol, sec-butanol, tert-butanol, and 1-methoxypropan-2-ol, as well as toluene, xylenes, and veratrole. Particular preference is given to using ethanol, n-butanol, i-propanol, 1-methoxypropan-2-ol, toluene, xylenes, and veratrole. Particular preference is given to using ethanol, n-butanol, and xylenes. BCS221003-Ausland Dr. ML 68 The reaction according to step 3 is usually carried out at temperatures of 60-140 °C. The reaction is preferably carried out at 80-120 °C. The reaction is particularly preferably carried out at 90-110 °C. The carbamates (II), where R = MeO, EtO, iPrO, and n-BuO, are novel and also form part of the present invention. The following examples illustrate the invention in more detail.Terrific Broth (TB) culture media were prepared in demineralized water using 47.6 g / L granulated medium and 4 ml / L glycerol and sterilized at 121°C for 20 minutes. Cloning of Lipases Nucleotide sequences encoding lipases and lipase variants as described herein can be synthesized as known in the art, e.g., as offered by appropriate service providers such as Eurofins Genomics GmbH (Eurofins Genomics GmbH, Anzinger Str. 7a, 85560 Ebersberg, Germany). Briefly, nucleic acid sequences of wild-type lipase (SEQ ID No. 2) or related variants, as described herein, were cloned into an expression vector based on the pKA81a vector. Genetic elements were introduced into the modified pKA81a vector using well-known methods. For the expression of wild-type lipase orLipase variants were introduced into the expression vectors in electrocompetent MG1655 cells of Escherichia coli. Generation of enzyme variants Nucleotide substitutions (replacements) were introduced into the nucleic acid parent sequences, e.g., to achieve an amino acid exchange with other amino acids. Several molecular biological methods can be used to achieve these replacements. A useful method for producing a mutated nucleic acid and the corresponding mutated protein of the invention is site-directed mutagenesis at codons encoding one or more amino acids that are selected in advance. The methods for achieving these site-directed mutations are well known to those of ordinary skill in the art and are adequately described in the literature (in particular: Directed Mutagenesis: A Practical Approach, 1991, edited by M.J. McPherson, IRL PRESS) or are methods for which commercial kits (e.g.,The QUIKCHANGE™ Lightning Mutagenesis Kit from Qiagen or Stratagene can be used. After site-directed mutagenesis, nucleic acids were transformed into Escherichia coli MG1655 cells. Transformed cells were tested in appropriate biotransformation reactions to determine product yield and selectivity. Suitable biotransformation reactions are described below. Sequence verification was performed as is known in the art. Glycerol stocks of the E. coli cultures transformed with the respective expression plasmids were prepared by adding one volume of a 40% glycerol solution to one volume of an E. coli culture. To isolate single bacterial colonies, appropriate dilutions of E. coli cultures were plated on LB agar plates containing appropriate concentrations of kanamycin and incubated at 37°C until single colonies were obtained.Synthesis of ethyl [(1R,2S)-2,6-dimethyl-2,3-dihydro-1H-inden-1-yl]carbamate Example 1: A mixture of 10 g of 2,6-dimethylindan-1-amine (racemic mixture of 82% trans and 18% cis isomer), 2 eq. of diethyl carbonate, 0.3 mol% palladium catalyst (5% palladium on aluminum oxide), and 3 wt.% protein with lipase activity was charged into a 100 mL autoclave. The autoclave was sealed, and argon was applied and released three times at 5 bar each. Subsequently, hydrogen was applied and the mixture was stirred at 120 °C for 16 hours. The autoclave was then cooled to room temperature and released. The reaction mixture was diluted with diethyl carbonate and ethyl acetate and analyzed by HPLC. The conversion of 2,6-dimethyl-1-aminoindane and the chemoselectivity and stereoselectivity of the formation of ethyl [(1R,2S)-2,6-dimethyl-2,3-dihydro-1H-inden-1-yl]carbamate were determined (Table 5, entry 1).Examples 2 to 36 were carried out analogously to Example 1 (Table 5). Table 5:. BCS221003-Abroad Dr. ML 70 Example 38: A mixture of 150 g of 2,6-dimethylindan-1-amine (racemic mixture of 82% trans and 18% cis isomers), 2 eq. of diethyl carbonate, 0.3 mol% palladium catalyst (5% palladium on aluminum oxide), and 3 wt.% proteins with lipase activity was charged into a 1000 mL autoclave. The autoclave was sealed, and argon was applied and released three times, each at 5 bar. Subsequently, 3 bar of hydrogen was applied, and the mixture was stirred at 120 °C for 25 hours. The autoclave was then cooled to room temperature and released. The reaction mixture was diluted with a total of 578 g of diethyl carbonate and 350 mL of ethyl acetate and filtered with suction at 70 °C. The solution was concentrated under reduced pressure and the residue was analyzed.The conversion of 2,6-dimethyl-1-aminoindane and the chemoselectivity and stereoselectivity of the formation of ethyl [(1R,2S)-2,6-dimethyl-2,3-dihydro-1H-inden-1-yl]carbamate were determined by HPLC. Conversion: 99.7%, chemoselectivity: 93%, stereoselectivity: 95.6%. The purity and yield of ethyl [(1R,2S)-2,6-dimethyl-2,3-dihydro-1H-inden-1-yl]carbamate were determined by quantitative chromatography. 1 Determined by H NMR. Purity: 67.1%, yield 86.9% of theory. 1 H-NMR (400 MHz; CDCl3) δ = 7.08–7.01 (m, 3H), 4.81–4.71 (m, 2H), 4.19 (q, J = 8.0 Hz, 2H), 3.00 (dd, J = 8.0, 14.0 Hz, 1H), 2.51–2.45 (m, 1H), 2.33 (s, 3H), 2.16 (dt, J = 8.0, 14.0 Hz, 1H), 1.31–1.26 (m, 6H). The derivative methyl [(1R,2S)-2,6-dimethyl-2,3-dihydro-1H-inden-1-yl]carbamate was prepared analogously using dimethyl carbonate and was obtained as a white solid. 1H-NMR (400 MHz; CDCl3) δ = 7.08-7.01 (m, 3H), 4.79-4.78 (m, 2H), 3.74 (s, 3H), 3.00 (dd, J = 8.0, 16.0 Hz, 1H), 2.51-2.45 (m, 1H), 2.32 (s, 3H), 2.20-2.10 (m, 1H), 1.27 (d, J = 8.0 Hz, 3H).

Claims

BCS221003-Ausland Dr. ML 72 Patent claims:

1. Process for the preparation of almost enantiomerically pure (1R,2S)-2,6-dimethyl-1-indanamine, characterized in that 1. in a first step, a mixture of the four stereoisomers of 2,6-dimethyl-1-indanamine (I) is reacted with an acylating or carboxylating agent RC(=O)R 1 a) in the presence of a protein with the activity of a lipase, the mixture (III) is selectively converted into the corresponding amide or carbamate (II) and a mixture (III) of the unreacted stereoisomers of 2,6-dimethyl-1-indanamine, and b) the mixture (III) is isomerized in the presence of a metal catalyst and under hydrogen pressure at the same time as the biocatalytic conversion to the starting material of the four stereoisomers of 2,6-dimethyl-1-indanamine (I): M etallkatalysator, wherein the protein is encoded by an amino acid sequence selected from the group consisting of I. proteins which have at least 80% identity to the amino acid sequence shown under SEQ ID No. 1, II. proteins which have at least 80% identity to the amino acid sequence shown under SEQ ID No. 1 except that the amino acid sequence has a modification selected from the group consisting of i. the amino acid at position 186 is different from L; ii. the amino acid at position 280 is different from L; iii. the amino acid at position 312 is different from P; iv. the amino acid at position 3 is different from M; v. the amino acid at position 29 is different from N; vi. the amino acid at position 17 is different from L; vii. the amino acid at position 4 is different from S; BCS221003-Abroad Dr. ML 73 viii. the amino acid at position 18 is different from V; ix. the amino acid at position 202 is different from A; x. the amino acid at position 301 is different from D; xi. the amino acid at position 309 is different from P; xii. the amino acid at position 31 is different from Q; xiii. the amino acid at position 111 is different from Q; xiv. the amino acid at position 85 is different from W; xv. the amino acid at position 8 is different from K; xvi. the amino acid at position 79 is different from E; xvii. the amino acid at position 40 is different from K; 2. that in a second step the amide or carbamate (II) is separated from the secondary components by crystallization, and 3. that in a third step the amide or carbamate (II) is converted using a base or an acid to the almost enantiomerically pure (1R,2S)-2,6-dimethyl-1-indanamine (IV), wherein R is a radical from the group CH2OCH3, CH2OCH2CH3, CH3, OCH3, OCH2CH3, OCH(CH3)2, OCH2CH2CH2CH3, and wherein R 1 a radical from the group OCH3, OCH2CH3, OCH(CH3)2 and OCH2CH2CH2CH3.

2. The process according to claim 1, wherein R is a radical from the group OCH3 and OCH2CH3, and R 1 a residue from the group OCH3 and OCH2CH3. BCS221003-Ausland Dr. ML 74 3. Method according to claim 1 or 2, characterized in that the protein is selected from the group consisting of a) proteins which comprise the amino acid sequence shown in SEQ ID No. 1, apart from the fact that the amino acid at position 186 is different from L;b) proteins with an amino acid sequence with at least 80% identity to the amino acid sequence shown under a), provided that the amino acid at position 186 is different from L, wherein preferably the amino acid in the proteins according to a) or b) at position 186 is F, W, Y, E, D, Q, T, H, P, C, K, S, N, I or V.

4. The method according to claim 3, characterized in that the protein has at least one, preferably at least two, more preferably at least three further amino acid substitutions selected from the group consisting of (i) the amino acid at position 79 is different from E, preferably the amino acid at position 79 is S, W or I, more preferably the amino acid at position 79 is S; (ii) the amino acid at position 202 is different from A, preferably the amino acid at position 202 is N; (iii) the amino acid at position 280 is different from L, preferably the amino acid at position 280 is A;(iv) the amino acid at position 301 is different from D, preferably the amino acid at position 301 is A; (v) the amino acid at position 3 is different from M, preferably the amino acid at position 3 is Q; (vi) the amino acid at position 11 is different from C, preferably the amino acid at position 11 is A; (vii) the amino acid at position 17 is different from L, preferably the amino acid at position 17 is P; (viii) the amino acid at position 40 is different from K, preferably the amino acid at position 40 is M;(ix) the amino acid at position 111 is different from Q, preferably the amino acid at position 111 is E.

5. The method according to claim 3 or 4, characterized in that the protein has the amino acid substitutions at position 186 and at position 79 and at position 301 and at position 40, wherein preferably the amino acid at position 186 is Y and the amino acid at position 79 is S and the amino acid at position 301 is A and the amino acid at position 40 is M.; BCS221003-Ausland Dr. ML 75 6. The process according to any one of claims 1 to 5, characterized in that the protein is used in an amount of 0.1-50 wt.% based on the mixture (I).

7. The process according to any one of claims 1 to 6, characterized in that the protein is used in an amount of 0.5-10 wt.% based on the mixture (I).

8. The process according to any one of claims 1 to 7, characterized in that the protein is used in an amount of 1-5 wt.% based on the mixture (I).

9. The process according to any one of claims 1 to 8, characterized in that step 1 is carried out without solvent or in the presence of a solvent from the group consisting of toluene, xylenes, mesitylene, n-butanol, and ethanol.

10. The process according to any one of claims 1 to 9, characterized in that in step 1, the acylating or carboxylating agent RC(=O)R 1preferably in an amount of 1-25 equivalents based on the amount of substance used in the mixture (I).

11. The process according to any one of claims 1 to 10, characterized in that in step 1 the acylating or carboxylating agent RC(=O)R 1preferably in an amount of 1-5 equivalents based on the amount of mixture (I) used.

12. The process according to any one of claims 1 to 11, characterized in that in step 1, the reaction is carried out at temperatures of 70-130°C.

13. The process according to any one of claims 1 to 12, characterized in that in step 1, the reaction is carried out at temperatures of 105-125°C.

14. The process according to any one of claims 1 to 13, characterized in that in step 1, a palladium on carbon (Pd / C) or a palladium on aluminum oxide (Pd / Al2O3) catalyst with a palladium loading of 0.5-10 wt. % is used.

15. The process according to claim 14, wherein the catalysts are used in an amount of 0.5-5 wt. % based on the compounds of formula (I). 16.Process according to one of claims 1 to 15, characterized in that in step 1 the reaction is carried out in a pressure range of 1-5 bar hydrogen pressure. BCS221003-Ausland Dr. ML 76 17. Compounds of formula (II) where R = MeO, EtO, iPrO and n-BuO.

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