Modular optimization of multi-gene pathways for fatty acids production in E. coli

Peng Xu, Qin Gu, Wenya Wang, Lynn Wong, Adam G.W. Bower, Cynthia H. Collins and Mattheos A.G. Koffas ()
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Peng Xu: Rensselaer Polytechnic Institute
Qin Gu: Rensselaer Polytechnic Institute
Wenya Wang: Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute
Lynn Wong: Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute
Adam G.W. Bower: Rensselaer Polytechnic Institute
Cynthia H. Collins: Rensselaer Polytechnic Institute
Mattheos A.G. Koffas: Rensselaer Polytechnic Institute

Nature Communications, 2013, vol. 4, issue 1, 1-8

Abstract: Abstract Microbial fatty acid-derived fuels have emerged as promising alternatives to petroleum-based transportation fuels. Here we report a modular engineering approach that systematically removed metabolic pathway bottlenecks and led to significant titre improvements in a multi-gene fatty acid metabolic pathway. On the basis of central pathway architecture, E. coli fatty acid biosynthesis was re-cast into three modules: the upstream acetyl coenzyme A formation module; the intermediary acetyl-CoA activation module; and the downstream fatty acid synthase module. Combinatorial optimization of transcriptional levels of these three modules led to the identification of conditions that balance the supply of acetyl-CoA and consumption of malonyl-CoA/ACP. Refining protein translation efficiency by customizing ribosome binding sites for both the upstream acetyl coenzyme A formation and fatty acid synthase modules enabled further production improvement. Fed-batch cultivation of the engineered strain resulted in a final fatty acid production of 8.6 g l−1. The modular engineering strategies demonstrate a generalized approach to engineering cell factories for valuable metabolites production.

Date: 2013
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DOI: 10.1038/ncomms2425

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