Li Jin, Han Ruizhi, Xu Guochao, Dong Jinjun, Ni Ye
Wei Sheng Wu Xue Bao. 2015 Nov 4;55(11):1427-36.
] Several key genes (thlA, bcs-operon/crt-bcd1 -etfB2-fixB2-hbd and adhE) in butanol pathway from Clostridium saccharobutylicum DSM13864 were cloned, and a butanol-producing Escherichia coli strain was successfully constructed.
Using genome of Clostridium saccharobutylicum DSM13864 as template, the key genes in butanol synthesis pathway were amplified, the recombinant plasmids pETDuet-bcs and pRSFDuet-thlA-adhE were constructed. Then the resultant plasmids were transformed into E. coli JM109 (DE3) to obtain E. coli BUT1 for butanol production, under the semi-anaerobic condition. Effects of different mediums on butanol production were studied.
The recombinant E. coli was capable of producing butanol (25.4 mg/L) under semi-anaerobic fermentation. After optimization on the fermentation medium, butanol titer reached 34.1 mg/L.
Butanol production by recombinant E. coli harboring exogenous butanol-producing pathway from Clostridium saccharobutylicum provides a feasible solution to overcome the hurdles in traditional butanol production approach by Clostridia.
克隆了丙酮丁醇梭菌DSM13864丁醇合成途径中的几个关键基因（thlA、bcs操纵子/crt-bcd1 -etfB2-fixB2-hbd和adhE），并成功构建了一株产丁醇的大肠杆菌菌株。
以丙酮丁醇梭菌DSM13864的基因组为模板，扩增丁醇合成途径中的关键基因，构建重组质粒pETDuet-bcs和pRSFDuet-thlA-adhE。然后将所得质粒转化到大肠杆菌JM109（DE3）中，在半厌氧条件下获得用于生产丁醇的大肠杆菌BUT1。研究了不同培养基对丁醇生产的影响。
重组大肠杆菌在半厌氧发酵条件下能够产生丁醇（25.4 mg/L）。对发酵培养基进行优化后，丁醇产量达到34.1 mg/L。
携带来自丙酮丁醇梭菌的外源丁醇合成途径的重组大肠杆菌产丁醇为克服传统梭菌丁醇生产方法中的障碍提供了一种可行的解决方案。
