Improvement of catechin production in Escherichia coli through combinatorial metabolic engineering

doi:10.1016/j.ymben.2014.12.002

	Improvement of catechin production in Escherichia coli through combinatorial metabolic engineering
	Zhao, Shujuan 1,3; Jones, J. Andrew 1; Lachance, Daniel M.2; Bhan, Namita 1; Khalidi, Omar 1; Venkataraman, Sylesh 5; Wang, Zhengtao 3,4; Koffas, Mattheos A.G.1,2
刊名	Metabolic Engineering
	2015-03-01
卷号	28 页码:43-53
ISSN号	10967176
DOI	10.1016/j.ymben.2014.12.002
文献子类	Article
英文摘要	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 tea catechins in Escherichia coli, combinatorial strategies were employed using the ePathBrick vectors to express the committed catechin pathway: flavanone 3β-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 F3Hsynoriginally from Camellia sinensis, DFRsynfrom Anthurium andraeanum, C. sinensis, or Fragaria ananass, and LARsynfrom 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 DFRsynand LARsynenabling a titer of 374.6±43.6mg/L of (+)-catechin. Improving the NADPH availability via the δpgiδppc mutation, BLδpgiδppc-p148 produced the highest titer of catechin at 760.9±84.3mg/L. After utilizing a library of scaffolding proteins, the strain BLδpgiδppc-p168-759 reached the highest titer of (+)-catechin of 910.9±61.3mg/L from 1.0g/L of eriodictyol in batch culture with M9 minimal media. The impact of oxygen availability on the biosynthesis of catechin was also investigated.
语种	英语
出版者	Academic Press Inc.
内容类型	期刊论文
源URL	[http://119.78.100.183/handle/2S10ELR8/266978]
专题	中国科学院上海药物研究所
通讯作者	Koffas, Mattheos A.G.
作者单位	1.Department of Chemical and Biological Engineering, Rensselaer Polytechnic Institute, Troy; NY; 12180, United States; 2.Department of Biological Sciences, Rensselaer Polytechnic Institute, Troy; NY; 12180, United States; 3.The SATCM Key Laboratory for New Resources and Quality Evaluation of Chinese, Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, Shanghai; 201203, China; 4.The MOE Key Laboratory for Standardization of Chinese Medicines and Medicines, Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, Shanghai; 201210, China; 5.Chromadex Inc., 2830 Wilderness Place, Boulder; CO; 80301, United States
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.,Khalidi, Omar.,...&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.