Method for producing acetoin through high-efficiency fermentation by appropriately expressing novel bacillus subtilis NADH oxidizing enzyme

A technology of Bacillus subtilis and oxidase, which is applied in the fields of genetic engineering and fermentation engineering to achieve the effect of shortening the fermentation period
CN104017758AInactive Publication Date: 2014-09-03JIANGNAN UNIV
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Patent Information

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

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Abstract

The invention relates to a method for producing acetoin through high-efficiency fermentation by appropriately expressing a novel bacillus subtilis NADH oxidizing enzyme and belongs to the fields of genetic engineering and fermentation engineering. According to the method, the noval bacillus subtilis NADH oxidizing enzyme is found for the first time in genome of a high-yield acetoin bacterial strain B.subtilis JNA which is autonomously screened by a laboratory and has independent intellectual property, the enzymatic property of the noval bacillus subtilis NADH oxidizing enzyme is determined while an NADH oxidizing enzyme is expressed in bdhA gene deleted bacillus subtilis by appropriately expressing a carrier pMA5-PbdhA, finally the acetoin is produced through fermentation by utilizing recombinant bacteria modified by metabolic engineering, about 56.7g / L acetoin and 1.2g / L of 2,3-butanediol are obtained, the contents of 2,3-butanediol, lactic acid and acetic acid are respectively reduced by about 92.3%, 70.1% and 75.0%, the production efficiency of the acetoin is increased to 0.68g / (L.h), and a fermentation period is shortened for 1.7 times compared with the original bacterial strain; NADH regulation in bacillus subtilis is utilized for producing the acetoin for the first time at home and abroad, and the aims of shortening the fermentation period and reducing a NADH-dependent by-product are realized.
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Description

technical field

[0001] The method utilizes moderately expressed novel NADH oxidase to reduce NADH-dependent by-products and increase the yield of acetoin. The invention belongs to the fields of genetic engineering and fermentation engineering. It specifically relates to the screening of a novel NADH oxidase, the construction of genetically engineered bacteria and its fermentation production. technical background

[0002] Acetoin occurs naturally in corn, grapes, cocoa, apples, bananas, cheese, meat, and many other foods. It is a widely used and lovable food spice, with a strong creamy, fat, butter-like aroma, and a pleasant milky aroma after being highly diluted, so acetoin is mainly used to prepare milk-flavored, meat-flavored type, strawberry-flavored flavor, or directly used in dairy products. my country's national standard GB2760-86 clearly stipulates that it is a food-grade spice that is allowed to be used, and the safety number of the American Food and Extraction Ass...

Claims

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