Gene recombination bacterium and application thereof in preparing chiral pure acetoin and 2,3-butanediol

Abstract

The invention discloses an E. coli BL21(pETDuet-ydjLnox) containing a 2R,3R-butanediol dehydrogenase gene ydjL and an NADH oxidase gene nox, wherein the strain is preserved in the 'China Center for Type Culture Collection' on December 23 in 2008, and the preservation number is CCTCC NO: M 208259. The invention also discloses application of a gene recombination bacterium in producing chiral S-AC by catalyzing meso-BD, producing chiral R-AC by catalyzing 2R,3R-BD and producing chiral pure 2S,3S-BD by splitting a 2,3-BD mixture. The concentration of a chiral AC prepared from the recombinant E. coli can reach over 6 grams per liter (the ee value is more than or equal to 96 percent); and the ee value of the chiral pure 2S,3S-BD is more than or equal to 98 percent, and the regeneration of a cofactor is achieved, thus the gene recombination bacterium has great industrial application prospect.

Description

technical field

[0001] The present invention relates to a gene recombination bacterium and application thereof, specifically, relates to a recombination capable of combining 2R, 3R-butanediol dehydrogenase (2R, 3R-BDH) gene (ydjL) and NADH oxidase gene ( nox) co-expressed Escherichia coli, and use this engineering bacteria to efficiently transform meso-2, 3-butanediol (meso-BD) to produce S-acetoin (S-AC), 2R, 3R-butanediol (2R, 3R-BD) the method for producing R-acetoin (R-AC) and resolution 2,3-butanediol (2,3-BD) mixture produces 2S, 3S-butanediol (2S, 3S -BD) approach. Background technique

[0002] Acetoin (AC) is usually pale yellow liquid or crystal, has two isomers (R-AC and S-AC), naturally exists in wine, honey, cocoa, butter, coffee, strawberry and Rainbow currant and other substances. The national standard GB2760-86 stipulates that it is a food spice that is allowed to be used, and the FEMA safety number is 2008. AC also has very important uses in the food, fra...

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