MUTANT NITRILE HYDRATASE, NUCLEIC ACID WHICH ENCODES SAID MUTANT NITRILE HYDRATASE, EXPRESSION VECTOR AND TRANSFORMANT CONTAINING SAID NUCLEIC ACID, METHOD OF PRODUCING SAID MUTANT NITRILE HYDRATASE AND METHOD OF PRODUCING AMIDE COMPOUND Russian patent published in 2020 - IPC C12N15/09 C12N15/70 C12N5/10 

FIELD: biotechnology.

SUBSTANCE: group of inventions relates to biotechnology. Disclosed is a mutant nitrile hydratase derived from Pseudonocardia thermophila, having α sub-unit and β subunit and containing at least one amino acid residue substitution selected from the group consisting of the following amino acid residues (a) - (e): (a) replacing the amino acid corresponding to 40^th amino acid residue from the N-terminus of SEQ ID NO: 1, by Asn in α-subunit, (b) replacing an amino acid corresponding to 43^rd amino acid residue from the N-terminus of SEQ ID NO: 1, by Val in α-conjunct (c) replacing the amino acid corresponding to 205^th amino acid residue from the N-terminus of SEQ ID NO: 2, by Val in β-subunit, (d) replacing an amino acid corresponding to 206^th amino acid residue from the N-terminus of SEQ ID NO: 2, on Gln in β-solid and (e) replacing the amino acid corresponding to 215th amino acid residue from the N-terminus of SEQ ID NO: 2, by Asn in β-conjuncture. Also presented is a nucleic acid coding mutant nitrile hydratase, an expression vector containing nucleic acid, a transformant for producing mutant nitrile hydratase containing an expression vector. As well as a method of producing mutant nitrile hydratase, involving cultivation of a transformant in a medium and recovering mutant nitrile hydratase from at least one of the cultivated transformant or medium, and a method of producing an amide compound, involving bringing the mutant nitrile hydratase into contact with a nitrile compound and removing impurities from the solution containing the amide compound at pH from 3.5 to 6.5.

EFFECT: group of inventions provides obtaining mutant nitrile hydratase, which preserves excellent enzymatic activity catalyzing reaction of synthesis of amide compound from nitrile compound even in acid range at pH from 3_5 to 6_5.

14 cl, 24 tbl, 29 ex