Method for producing acetoin by efficient bioconversion of 2,3-butanediol by using Bacillus subtilis nicotinamide adenine dinucleotide (NAD)<+> regeneration system

A technology of acetoin and butanediol, applied in the fields of genetic engineering and bioengineering, can solve the problems of low accumulation concentration, difficult separation in downstream engineering, inability to obtain pure acetoin, etc., and achieves efficient production, cost reduction, and simplified separation Effect
CN104017764AInactive Publication Date: 2014-09-03JIANGNAN UNIV
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
JIANGNAN UNIV
Publication Date
2014-09-03
Estimated Expiration
Not applicable · inactive patent

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Abstract

The invention discloses a method for producing acetoin by efficient bioconversion of 2,3-butanediol by using a Bacillus subtilis nicotinamide adenine dinucleotide (NAD)<+> regeneration system, and belongs to the field of genetic engineering. Acetoin reductase and educed form of nicotinamide-adenine dinucleotide (NADH) oxidase in a high-yield acetoin strain B.subtilis JNA with independent intellectual property rights, which are autonomously screened from a laboratory are cloned by the method disclosed by the invention, and excessive coexpression of the acetoin reductase and NADH oxidase in B.subtilis JNA is carried out, so that the production of acetoin by effective conversion of the 2,3-butanediol in the wild-type high-yield acetoin bacillus subtilis by virtue of the NAD<+> regeneration system is realized for the first time at home and abroad. The enzyme activity determination and intracellular coenzyme level research on the built gene engineering strain prove that the 2,3-butanediol can be lastingly and effectively converted by B.subtilis JNA / pMA5-bdhA-yodC to produce the acetoin. 120g / L of 2,3-butanediol can be finally converted into about 92.5g / L of acetoin by the B.subtilis JNA / pMA5-bdhA-yodC when the temperature is 40 DEG C and the pH is 8.0 under the optimal whole-cell conversion condition of adding 5mM of MnC12, the acetoin yield can be up to 2.31g / (L.h), and is the highest lever for producing the acetoin from the bacillus subtilis in the current report, and a foundation is provided for industrial production of the acetoin from microorganisms.
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Description

technical field

[0001] Utilize NAD + The invention relates to a method for efficiently biotransforming 2,3-butanediol to produce acetoin in a regeneration system, and the invention belongs to the fields of genetic engineering and bioengineering. Specifically, it relates to the construction of a genetically engineered bacterial strain co-expressed with acetoin reductase and NADH oxidase and a method for using the bacterial strain to transform 2,3-butanediol into whole cells to produce acetoin. technical background

[0002] Acetoin has a wide range of applications in food flavoring, biochemistry and pharmacology. At present, there are relatively in-depth researches on its synthesis and production at home and abroad. In recent years, the production of acetoin by microbial fermentation has received more and more attention, and scientists from various countries have carried out work in this area one after another. At present, the research on the production of acetoin by ferment...

Claims

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