Chu Haipei, Xin Bo, Liu Peihai, Wang Yu, Li Lixiang, Liu Xiuxiu, Zhang Xuan, Ma Cuiqing, Xu Ping, Gao Chao
State Key Laboratory of Microbial Technology, Shandong University, Jinan, 250100 People's Republic of China.
Rizhao Entry-Exit Inspection and Quarantine Bureau, Rizhao, 276800 People's Republic of China.
Biotechnol Biofuels. 2015 Sep 15;8:143. doi: 10.1186/s13068-015-0324-x. eCollection 2015.
Butane-2,3-diol (2,3-BD) is a fuel and platform biochemical with various industrial applications. 2,3-BD exists in three stereoisomeric forms: (2R,3R)-2,3-BD, meso-2,3-BD and (2S,3S)-2,3-BD. Microbial fermentative processes have been reported for (2R,3R)-2,3-BD and meso-2,3-BD production.
The production of (2S,3S)-2,3-BD from glucose was acquired by whole cells of recombinant Escherichia coli coexpressing the α-acetolactate synthase and meso-butane-2,3-diol dehydrogenase of Enterobacter cloacae subsp. dissolvens strain SDM. An optimal biocatalyst for (2S,3S)-2,3-BD production, E. coli BL21 (pETDuet-PT7-budB-PT7-budC), was constructed and the bioconversion conditions were optimized. With the addition of 10 mM FeCl3 in the bioconversion system, (2S,3S)-2,3-BD at a concentration of 2.2 g/L was obtained with a stereoisomeric purity of 95.0 % using the metabolically engineered strain from glucose.
The engineered E. coli strain is the first one that can be used in the direct production of (2S,3S)-2,3-BD from glucose. The results demonstrated that the method developed here would be a promising process for efficient (2S,3S)-2,3-BD production.
2,3-丁二醇(2,3-BD)是一种具有多种工业应用的燃料和平台生物化学品。2,3-丁二醇以三种立体异构体形式存在:(2R,3R)-2,3-丁二醇、内消旋-2,3-丁二醇和(2S,3S)-2,3-丁二醇。已有关于微生物发酵生产(2R,3R)-2,3-丁二醇和内消旋-2,3-丁二醇的报道。
通过共表达阴沟肠杆菌亚种溶解菌株SDM的α-乙酰乳酸合酶和内消旋-丁烷-2,3-二醇脱氢酶的重组大肠杆菌全细胞,实现了从葡萄糖生产(2S,3S)-2,3-丁二醇。构建了用于生产(2S,3S)-2,3-丁二醇的最佳生物催化剂大肠杆菌BL21(pETDuet-PT7-budB-PT7-budC),并优化了生物转化条件。在生物转化体系中添加10 mM FeCl3,使用代谢工程菌株从葡萄糖中获得了浓度为2.2 g/L、立体异构体纯度为95.0%的(2S,3S)-2,3-丁二醇。
工程化大肠杆菌菌株是首个可用于直接从葡萄糖生产(2S,3S)-2,3-丁二醇的菌株。结果表明,本文开发的方法将是一种高效生产(2S,3S)-2,3-丁二醇的有前景的工艺。