title: Mutation breeding of acetoin high producing Bacillus subtilis blocked in 2,3-butanediol dehydrogenase.
abstract: Bacillus subtilis mutants were obtained after the wild strain JNA 3-10 was mutagenized by UV irradiation coupled with diethyl sulfate. A visual filter assay was employed for the qualitative identification of 2,3-butanediol dehydrogenase (BDH) blocked B. subtilis. Selected mutants were tested for the activities of acetoin reductase (AR) and BDH. According to further batch fermentation, one mutant named JNA-UD-6 that produced 24.3 % more acetoin than JNA 3-10 with the corresponding byproducts of 2,3-butanediol decreased by 39.8 % was isolated. A nonsense mutation (p.Tyr118X) that precluded the synthesis of a full-length functional AR/BDH within the bdhA gene of JNA-UD-6 was detected. Acetoin production of JNA-UD-6 was further improved to about 53.9 g/L in a 5-L fermentor with 150 g/L glucose consumed. However,a small amount of 2,3-butanediol was found in late phase of JNA-UD-6 fermentation, and it was due to the existence of a putative gene that encoding a minor AR. This work proved a strategy to efficiently breeding an acetoin high producing strain by traditional mutation methods.
authors.affiliation: The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, 214122, Jiangsu, People's Republic of China.
authors.forename: Xian
authors.initials: X
authors.lastname: Zhang
authors.forename: Rongzhen
authors.initials: R
authors.lastname: Zhang
authors.forename: Taowei
authors.initials: T
authors.lastname: Yang
authors.forename: Jing
authors.initials: J
authors.lastname: Zhang
authors.forename: Meijuan
authors.initials: M
authors.lastname: Xu
authors.forename: Huazhong
authors.initials: H
authors.lastname: Li
authors.forename: Zhenghong
authors.initials: Z
authors.lastname: Xu
authors.forename: Zhiming
authors.initials: Z
authors.lastname: Rao
mesh_terms: D000093:Acetoin / Q000378:metabolism*; D000429:Alcohol Oxidoreductases / Q000235:genetics*; D001412:Bacillus subtilis / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism* / Q000528:radiation effects; D002072:Butylene Glycols / Q000378:metabolism; D018389:Codon, Nonsense; D055786:Gene Knockout Techniques; D060847:Metabolic Engineering / Q000379:methods*; D016296:Mutagenesis; D009153:Mutagens / Q000378:metabolism; D050505:Mutant Proteins / Q000235:genetics; D014466:Ultraviolet Rays
chemical_list: D002072:Butylene Glycols; D018389:Codon, Nonsense; D009153:Mutagens; D050505:Mutant Proteins; C026978:2,3-butylene glycol; D000093:Acetoin; D000429:Alcohol Oxidoreductases; C019801:butanediol dehydrogenase
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