RU2003108867A - BUTINOL I EXTERASIS - Google Patents

Claims (20)

1. A protein having butynol I esterase activity, containing at least one partial amino acid sequence corresponding to sequence number 3, 4, 5 or 6, as well as its functional equivalents having butynol I esterase activity. 2. The protein according to claim 1, containing the amino acid sequence corresponding to sequence number 2, as well as its functional equivalents. 3. The protein according to claim 1 or 2, wherein the functional equivalent contains at least one partial sequence of at least 10 consecutive amino acid residues from the amino acid sequence corresponding to sequence number 2, 3, 4, 5, or 6. 4. The protein according to one of the preceding paragraphs, containing a polypeptide chain with a molecular weight of about 41300 Da. 5. The protein according to one of the preceding paragraphs, characterized in that it is produced by the strain Pseudomonas glumae Lu 2023 under the number DSM 13176. 6. The protein according to one of the preceding paragraphs, characterized in that it catalyzes at least one of the following reactions: a) enantio-selective hydrolysis of optically active esters of the formula I

where R ¹ means a normal or branched, if necessary, once or repeatedly substituted alkyl with 1-10 carbon atoms, alkenyl with 2-10 carbon atoms, alkynyl with 2-10 carbon atoms, and R ² means normal or branched, if necessary, singly or repeatedly substituted alkyl with 1-10 carbon atoms, alkenyl with 2-10 carbon atoms, alkynyl with 2-10 carbon atoms, C ₇ -C ₁₅ aralkyl or containing one or more nuclei, optionally, once or repeatedly substituted aromatic residue, R ¹ and / or R ² include ce I at least one asymmetric carbon atom; and b) enantio-selective transesterification of esters of formula I with optically active alcohols of formula II

where R ² has the above meaning, and, if necessary, contains one or more asymmetric carbon atoms. 7. A polynucleotide encoding a protein according to one of the preceding paragraphs, as well as its functional equivalents, complementary polynucleotides to it, and nucleotide sequences hybridizing with it. 8. The polynucleotide according to claim 7, containing a partial nucleotide sequence of at least 30 consecutive nucleotide residues of nucleotide sequence number 1. 9. The expression block containing at least one polynucleotide according to claim 7 or 8, operatively associated with at least one regulatory nucleotide sequence. 10. Recombinant vector for transformation of a eukaryotic or prokaryotic host organism containing the polynucleotide according to claim 7 or 8, or an expression block according to claim 9. 11. The method of producing the protein according to one of claims 1 to 6, in which a microorganism is cultured endogenously producing the protein or transformed with the vector of claim 10 and the protein is isolated from the cell culture. 12. The method according to claim 11, in which the strain Pseudomonas glumae Lu 2023 under the number DSM 13176 is used as the producing microorganism. 13. The protein obtained by the method according to one of claim 11 or 12. 14. The strain Pseudomonas glumae Lu 2023 under the number DSM 13176, as well as its variants and mutants. 15. A microorganism, characterized in that it carries a vector according to claim 10. 16. The method of enantio-selective hydrolysis of esters using the protein according to one of claims 1 to 6, in which a) the protein is contacted with a mixture of stereoisomers of an optically active ester of formula I; and b) the optically active compounds and / or non-hydrolyzed ether enantiomers resulting from stereo-selective hydrolysis of one of the stereoisomers are isolated from the reaction solution. 17. A method of enantio-selective transesterification, in which a) a mixture of stereoisomers of an optically active alcohol of formula II is contacted with an ester of formula I in the presence of a protein according to one of claims 1 to 6, and the unreacted stereoisomer of alcohol is isolated from the reaction solution; or b) the mixture of stereoisomers of an optically active ester of formula I is contacted with an alcohol of formula II in the presence of a protein according to one of claims 1 to 6, and one of the stereoisomers of the optically active alcohol contained in the ether is isolated from the reaction solution. 18. The method according to 17, in which transesterification uses vinyl ether as an acylating agent for optically active alcohol. 19. The method according to one of claims 16, 17 or 18, wherein an organic solvent is used as the reaction solution. 20. Optically active alcohols, carboxylic acids or esters obtained using the protein according to one of claims 1 to 6.