Modular Pathway Engineering of Diterpenoid Synthases and the Mevalonic Acid Pathway for Miltiradiene Production

	Modular Pathway Engineering of Diterpenoid Synthases and the Mevalonic Acid Pathway for Miltiradiene Production
	Zhou, Yongjin J.1,6; Gao, Wei 3,4; Rong, Qixian 3; Jin, Guojie 1,6; Chu, Huiying 2; Liu, Wujun 1; Yang, Wei 1; Zhu, Zhiwei 1,6; Li, Guohui 2; Zhu, Guofeng 5
刊名	journal of the american chemical society
	2012-02-15
卷号	134 期号:6 页码:3234-3241
ISSN号	0002-7863
通讯作者	huangluqi
产权排序	1,1
英文摘要	microbial production can be advantageous over the extraction of phytoterpenoids from natural plant sources, but it remains challenging to rationally and rapidly access efficient pathway variants. previous engineering attempts mainly focused on the mevalonic acid (mva) or methyl-d-erythritol phosphate (mep) pathways responsible for the generation of precursors for terpenoids biosynthesis, and potential interactions between diterpenoids synthases were unexplored. miltiradiene, the product of the stepwise conversion of (e,e,e)-geranylgeranyl diphosphate (ggpp) catalyzed by diterpene synthases smcps and smksl, has recently been identified as the precursor to tanshionones, a group of abietane-type norcliterpenoids rich in the chinese medicinal herb salvia miltiorrhiza. here, we present the modular pathway engineering (mope) strategy and its application for rapid assembling synthetic miltiradiene pathways in the yeast saccharomyces cerevisiae. we predicted and analyzed the molecular interactions between smcps and smksl, and engineered their active sites into close proximity for enhanced metabolic flux channeling to miltiradiene biosynthesis by constructing protein fusions. we show that the fusion of smcps and smksl, as well as the fusion of bts1 (ggpp synthase) and erg20 (farnesyl diphosphate synthase), led to significantly improved miltiradiene production and reduced byproduct accumulation. the mope strategy facilitated a comprehensive evaluation of pathway variants involving multiple genes, and, as a result, our best pathway with the diploid strain yj2x reached miltiradiene titer of 365 mg/l in a 15-l bioreactor culture. these results suggest that terpenoids synthases and the precursor supplying enzymes should be engineered systematically to enable an efficient microbial production of phytoterpenoids.
学科主题	物理化学
WOS标题词	science & technology ; physical sciences
类目[WOS]	chemistry, multidisciplinary
研究领域[WOS]	chemistry
关键词[WOS]	saccharomyces-cerevisiae ; metabolic enzymes ; yeast ; overproduction ; expression ; reductase ; biosynthesis ; organization ; complexes ; coli
收录类别	SCI
语种	英语
WOS记录号	WOS:000301161500065
公开日期	2013-10-11
内容类型	期刊论文
源URL	[http://159.226.238.44/handle/321008/117888]
专题	大连化学物理研究所_中国科学院大连化学物理研究所
作者单位	1.Chinese Acad Sci, Dalian Inst Chem Phys, Div Biotechnol, Dalian 116023, Peoples R China 2.Chinese Acad Sci, Dalian Inst Chem Phys, State Key Lab Mol React Dynam, Dalian 116023, Peoples R China 3.CACMS, Inst Chinese Mat Med, Beijing 100700, Peoples R China 4.Capital Med Univ, Sch Tradit Chinese Med, Beijing 100069, Peoples R China 5.CAMS, Inst Pathogen Biol, Beijing 100730, Peoples R China 6.Chinese Acad Sci, Grad Univ, Beijing 100049, Peoples R China
推荐引用方式 GB/T 7714	Zhou, Yongjin J.,Gao, Wei,Rong, Qixian,et al. Modular Pathway Engineering of Diterpenoid Synthases and the Mevalonic Acid Pathway for Miltiradiene Production[J]. journal of the american chemical society,2012,134(6):3234-3241.
APA	Zhou, Yongjin J..,Gao, Wei.,Rong, Qixian.,Jin, Guojie.,Chu, Huiying.,...&Zhao, Zongbao K..(2012).Modular Pathway Engineering of Diterpenoid Synthases and the Mevalonic Acid Pathway for Miltiradiene Production.journal of the american chemical society,134(6),3234-3241.
MLA	Zhou, Yongjin J.,et al."Modular Pathway Engineering of Diterpenoid Synthases and the Mevalonic Acid Pathway for Miltiradiene Production".journal of the american chemical society 134.6(2012):3234-3241.