Novel (R)-2,3-butanediol dehydrogenase
AssigneeDaicel Chemical Industries, Ltd.
InventorsHiroaki Yamamoto, Keiko Onodera, Yoshiki Tani
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Abstract
The objective of the present invention is to produce a novel (R)-2,3-butanediol dehydrogenase useful for the production of ketones, alcohols, and particularly optically active vicinal diols. It has been found that Pichia angusta produces novel (R)-2,3-butanediol dehydrogenase that shows the high activity and high stereoselectivity. Further, polynucleotide encoding this (R)-2,3-butanediol dehydrogenase was cloned, and the nucleotide sequence thereof was determined. The expression of the glycerol dehydrogenase was carried out in heterologous microorganisms.
Claims23
An (R)-2,3-butanediol dehydrogenase, wherein
- (a) the dehydrogenase produces (R)-acetoin by acting on (2R,3R)-2,3-butanediol using nicotinamide adenine dinucleotide as a coenzyme and produces (2R,3R)-2,3-butanediol by reducing 2,3-butanedione using reduced form of nicotinamide adenine dinucleotide as a coenzyme;
- (b) the dehydrogenase uses nicotinamide adenine dinucleotide as a coenzyme in oxidation reaction and uses reduced form of nicotinamide adenine dinucleotide as a coenzyme in reduction reaction and preferentially oxidizes a hydroxyl group of 2,3-butanediol in (R) configuration;
- and
- (c) the dehydrogenase has 100 U or higher of (R)-2,3-butanediol dehydrogenase activity per 1 mg of the dehydrogenase when purified.
An isolated polynucleotide selected from the group consisting of:
- (a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO:1;
- (b) a polynucleotide encoding a polypeptide comprising the amino acid sequence of SEQ ID NO:2;
- (c) a polynucleotide encoding a polypeptide that comprises an amino acid sequence comprising the amino acid sequence of SEQ ID NO:
- 2 in which one or more amino acids are substituted, deleted, inserted, and/or added and that is functionally equivalent to a polypeptide comprising the amino acid sequence of SEQ ID NO:2;
- and
- (d) a polynucleotide that hybridizes under stringent conditions to a polynucleotide comprising the nucleotide sequence of SEQ ID NO:
- 1 and that encodes a polypeptide functionally equivalent to a polypeptide comprising the amino acid sequence of SEQ ID NO:2.
Dependencies are inferred from the claim text.