Alcohol dehydrogenase mutant and application thereof in synthesis of chiral diaryl alcohol compounds

Abstract

The invention discloses an alcohol dehydrogenase mutant and application thereof in synthesis of chiral diaryl alcohol compounds. The alcohol dehydrogenase mutant disclosed by the invention is a protein, which is obtained through mutating amino acid residues located in a catalytic center pocket of alcohol dehydrogenase TbSADH, and particularly is the protein, which is obtained through mutating 39thand / or 42th and / or 84th and / or 85th and / or 86th and / or 104th and / or 110th and / or 294th amino acid residues of an amino acid sequence of the alcohol dehydrogenase TbSADH. According to the alcohol dehydrogenase mutant and the application thereof, the alcohol dehydrogenase TbSADH is subjected to enzyme reforming by applying an orthogenesis technology and method, a series of mutants, which have enzyme activity to diaryl ketones with represented by (4-chlorophenyl)pyrid-2-ketone, are obtained, and the mutants are applied to biocatalytic synthesis of optically-pure chiral diaryl alcohol compounds.

Description

technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to alcohol dehydrogenase mutants obtained by enzymatic modification of alcohol dehydrogenase derived from Thermoanaerobacter brockii by applying directed evolution technology and methods, and these mutants are used as biocatalysts in Application of asymmetric reduction in the preparation of optically pure chiral bis-aryl alcohol compounds. Background technique [0002] As an important class of building blocks, chiral bisaryl secondary alcohols can be used in the synthesis of a variety of chiral drugs, and the process of synthesizing chiral bisaryl alcohols from prochiral bisaryl ketones through asymmetric reduction has atomic Advantages of high economy. Among them, (S)-(4-chlorophenyl)pyridine-2-methanol is a chiral precursor for the synthesis of antihistamines, which can be used to synthesize antihistamines carbinoxamine (Barouh et al., J. Med.Chem., 1971,14(9):834-836) an...

AI Technical Summary

Problems solved by technology

However, the efficiency of these whole-cell transformation methods is low, and there is no report on the amino acid sequence of the relevant enzyme

In 2007, Truppo et al. reported the use of commercial carbonyl reductase (ketoreductase, KRED) to asymmetrically reduce prochiral ketones to synthesize (S)-(4-chlorophenyl)pyridine-2-methanol, but the ee value was only 60 %, the sequence of the enzyme protein is unknown, and the source of the species has not been reported (Truppo et al., Org. Lett., 2007, 9(2):335-338)

However, since (S)-(4-chlorophenyl)pyridine-2-methanol is the precursor required for the synthesis of drug intermediates, there is currently no good enzyme that can efficiently catalyze (4-chlorophenyl)pyridine-2-methanone Generates (S)-products with high stereoselectivity

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