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Improvement of catechin production in Escherichia coli through combinatorial metabolic engineering 期刊论文

编号：

b919952b-aa6c-4208-85a4-f764e958a8fe
作者：

Zhao, Shujuan(赵淑娟)#^[1,3]Jones, J. Andrew#^[1]Lachance, Daniel M.^[2];Bhan, Namita^[1];Khalidi, Omar^[1];Venkataraman, Sylesh^[5];Wang, Zhengtao(王峥涛)^[3,4]Koffas, Mattheos A. G.*^[1,2]
语种：

英文
期刊：

METABOLIC ENGINEERING ISSN：1096-7176 2015 年 28 卷 (43 - 53) ; MAR
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关键词：

Catechin ePathBrick Copy number balancing NADPH availability Protein scaffolds Pathway optimization
摘要：

Reconstruction of highly efficient biosynthesis pathways is essential for the production of valuable plant secondary metabolites in recombinant microorganisms. In order to improve the titer of green Lea catechins in Escherichia coli, combinatorial strategies were employed using the ePathBrick vectors to express the committed catechin pathway: flavanone 3 beta-hydroxylase (F3H), dihydroflavonol 4-reductase (DFR), and leucoanthocyanidin reductase (LAR). Three F3H, three DFR, and two LAR genes originating from different plant species were selected and synthesized, to create 18 pathway variants to be screened in E. coli. Constructs containing F3H(syn) originally from Camellia sinensis, DFRsyn from Anthurium andraeanum, C. sinensis, or Fragaria ananass, and LAR(syn) from Desmodium uncinatum (p148, p158 and p168) demonstrated high conversion efficiency with either eriodictyol or naringenin as substrate. A highly efficient construct was created by assembling additional copies of DFRsyn and LAR(syn) enabling a titer of 374.6 +/- 43.6 mg/L of (+)-catechin. Improving the NADPH availability via Delta pgi Delta ppc mutation, BL Delta pgi Delta ppc-p148 produced the highest titer of catechin at 760.9 +/- 84.3 mg/L. After utilizing a library of scaffolding proteins. the strain BL Delta pgi Delta ppc-p168-759 reached the highest titer of (+)-catechin of 910.9 +/- 61.3 mg/L from 1.0 g/L of eriodictyol in batch culture with M9 minimal media. The impact of oxygen availability on the biosynthesis of catechin was also investigated. (C) 2014 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.
推荐引用方式
GB/T 7714：

Zhao Shujuan,Jones J. Andrew,Lachance Daniel M., et al. Improvement of catechin production in Escherichia coli through combinatorial metabolic engineering [J].METABOLIC ENGINEERING,2015,28:43-53.
APA：

Zhao Shujuan,Jones J. Andrew,Lachance Daniel M.,Bhan Namita,&Koffas Mattheos A. G..(2015).Improvement of catechin production in Escherichia coli through combinatorial metabolic engineering .METABOLIC ENGINEERING,28:43-53.
MLA：

Zhao Shujuan, et al. "Improvement of catechin production in Escherichia coli through combinatorial metabolic engineering" .METABOLIC ENGINEERING 28(2015):43-53.
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