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首页> 外文期刊>Applied Microbiology >Driving Forces Enable High-Titer Anaerobic 1-Butanol Synthesis in Escherichia coli
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Driving Forces Enable High-Titer Anaerobic 1-Butanol Synthesis in Escherichia coli

机译：驱动力可实现大肠杆菌中的高滴度厌氧1-丁醇合成

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摘要

1-Butanol, an important chemical feedstock and advanced biofuel, is produced by Clostridium species. Various efforts have been made to transfer the clostridial 1-butanol pathway into other microorganisms. However, in contrast to similar compounds, only limited titers of 1-butanol were attained. In this work, we constructed a modified clostridial 1-butanol pathway in Escherichia coli to provide an irreversible reaction catalyzed by trans -enoyl-coenzyme A (CoA) reductase (Ter) and created NADH and acetyl-CoA driving forces to direct the flux. We achieved high-titer (30 g/liter) and high-yield (70 to 88% of the theoretical) production of 1-butanol anaerobically, comparable to or exceeding the levels demonstrated by native producers. Without the NADH and acetyl-CoA driving forces, the Ter reaction alone only achieved about 1/10 the level of production. The engineered host platform also enables the selection of essential enzymes with better catalytic efficiency or expression by anaerobic growth rescue. These results demonstrate the importance of driving forces in the efficient production of nonnative products.
机译：1-丁醇是一种重要的化学原料和先进的生物燃料，是梭状芽胞杆菌产生的。已经进行了各种努力以将梭菌的1-丁醇途径转移到其他微生物中。但是，与相似的化合物相比，只能获得有限的1-丁醇滴度。在这项工作中，我们在大肠杆菌中构建了一条经修饰的梭菌1-丁醇途径，以提供一种由反式-烯酰辅酶A（CoA）还原酶（Ter）催化的不可逆反应，并产生了NADH和乙酰辅酶A驱动力来引导通量。我们厌氧地实现了1-丁醇的高滴度（30克/升）和高产率（理论值的70％到88％）的生产，可与甚至超过本地生产商证明的水平。如果没有NADH和乙酰辅酶A的驱动力，仅Ter反应就只能达到大约1/10的生产水平。经过设计的宿主平台还可以通过厌氧生长抢救来选择具有更好催化效率或表达能力的必需酶。这些结果证明了驱动力在有效生产非本地产品中的重要性。

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