Biotechnology and Bioengineering 2015-01-01

A synthetic O2 -tolerant butanol pathway exploiting native fatty acid biosynthesis in Escherichia coli.

András Pásztor, Pauli Kallio, Dávid Malatinszky, M Kalim Akhtar, Patrik R Jones

文献索引:Biotechnol. Bioeng. 112(1) , 120-8, (2014)



Several synthetic metabolic pathways for butanol synthesis have been reported in Escherichia coli by modification of the native CoA-dependent pathway from selected Clostridium species. These pathways are all dependent on the O2 -sensitive AdhE2 enzyme from Clostridium acetobutylicum that catalyzes the sequential reduction of both butyryl-CoA and butyraldehyde. We constructed an O2 -tolerant butanol pathway based on the activities of an ACP-thioesterase, acting on butyryl-ACP in the native fatty acid biosynthesis pathway, and a promiscuous carboxylic acid reductase. The pathway was genetically optimized by screening a series of bacterial acyl-ACP thioesterases and also by modification of the physical growth parameters. In order to evaluate the potential of the pathway for butanol production, the ACP-dependent butanol pathway was compared with a previously established CoA-dependent pathway. The effect of (1) O2 -availability, (2) media, and (3) co-expression of aldehyde reductases was evaluated systematically demonstrating varying and contrasting functionality between the ACP- and CoA-dependent pathways. The yield of butanol from the ACP-dependent pathway was stimulated by enhanced O2 -availability, in contrast to the CoA-dependent pathway, which did not function well under aerobic conditions. Similarly, whilst the CoA-dependent pathway only performed well in complex media, the ACP-dependent pathway was not influenced by the choice of media except in the absence of O2 . A combination of a thioesterase from Bacteroides fragilis and the aldehyde reductase, ahr, from E. coli resulted in the greatest yield of butanol. A product titer of ~300 mg/L was obtained in 24 h under optimal batch growth conditions, in most cases exceeding the performance of the reference CoA-pathway when evaluated under equivalent conditions.© 2014 Wiley Periodicals, Inc.


结构式 名称/CAS号 全部文献
甘油 结构式 甘油
无水氯化钙 结构式 无水氯化钙
正丁醇 结构式 正丁醇
磷酸氢二钾 结构式 磷酸氢二钾
硫酸镁 结构式 硫酸镁
磷酸二氢钾 结构式 磷酸二氢钾
氯化铵 结构式 氯化铵
二水氯化钙 结构式 二水氯化钙