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

TY - JOUR

T1 - Modular optimization of multi-gene pathways for fatty acids production in E. coli

AU - Xu, Peng

AU - Gu, Qin

AU - Wang, Wenya

AU - Wong, Lynn

AU - Bower, Adam G.W.

AU - Collins, Cynthia H.

AU - Koffas, Mattheos A.G.

PY - 2013

Y1 - 2013

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=84877804801&partnerID=8YFLogxK

U2 - 10.1038/ncomms2425

DO - 10.1038/ncomms2425

M3 - 文章

C2 - 23361000

AN - SCOPUS:84877804801

SN - 2041-1723

VL - 4

JO - Nature Communications

JF - Nature Communications

M1 - 1409

ER -