产丁醇大肠杆菌工程菌的构建

The Construction of Recombinant Escherichia coli Producing Butanol

【目的】为了在大肠杆菌(Escherichia coli)中导入改良的丁醇合成途径,使非生产菌株大肠杆菌具备产丁醇的能力。【方法】克隆大肠杆菌乙酰转移酶基因atoB和丙酮丁醇梭菌(Clostridium acetobutylicum)丁醇合成途径关键酶基因(crt、hbd、adhE),构建多顺反子表达质粒pSE380-atoB-adhE-crt-hbd;克隆齿垢密螺旋体(Treponema denticola)反式烯酰辅酶A还原酶基因ter,构建表达质粒pSTV29-ter,并将双质粒导入到大肠杆菌。【结果】构建的工程菌能半厌氧发酵产微量丁醇,产量为0.08g/L。【结论】大肠杆菌中的丁醇合成途径导入成功,构建了产丁醇的大肠杆菌工程菌。

[Objective] A modified 1-butanol pathway was constructed in non-native producer Escherichia coli to produce butanol. [Method]By cloning the acetyltransferase gene from E. coli, and the key butanol synthetic pathway genes crt,hbd and adhE from Clostridium acetobutylicurn , the polycistron expression plasmid pSE380- atoB - adhE - crt - hbd was constructed;by cloning the trans-2-enoyl-CoA reductase gene ter from Treponema denticola, the expression plasmid pSTV29-ter was constructed. [Result]The recombinant E. coli containing the double plasmids was fermented in micro-aerobic and the maximal concentration of butanol was 0.08g/L at 24h. [Conclusion]The butanol synthetic pathway was expressed in E. coli and the recombinant strain of producing butanol was constructed successfully.