Dissection of Malonyl-Coenzyme A Reductase of Chloroflexus aurantiacus Results in Enzyme Activity Improvement

CORC > 青岛生物能源与过程研究所 > 中国科学院青岛生物能源与过程研究所 > 材料生物技术研究中心

	Dissection of Malonyl-Coenzyme A Reductase of Chloroflexus aurantiacus Results in Enzyme Activity Improvement
	Liu, Changshui 1,3; Wang, Qi 1,3; Xian, Mo1,2 ; Ding, Yamei 4; Zhao, Guang 1,2
刊名	PLOS ONE
	2013-09-20
卷号	8 期号:9 页码:e75554
英文摘要	The formation of fusion protein in biosynthetic pathways usually improves metabolic efficiency either channeling intermediates and/or colocalizing enzymes. In the metabolic engineering of biochemical pathways, generating unnatural protein fusions between sequential biosynthetic enzymes is a useful method to increase system efficiency and product yield. Here, we reported a special case. The malonyl-CoA reductase (MCR) of Chloroflexus aurantiacus catalyzes the conversion of malonyl-CoA to 3-hydroxypropionate (3HP), and is a key enzyme in microbial production of 3HP, an important platform chemical. Functional domain analysis revealed that the N-terminal region of MCR (MCR-N; amino acids 1-549) and the C-terminal region of MCR (MCR-C; amino acids 550-1219) were functionally distinct. The malonyl-CoA was reduced into free intermediate malonate semialdehyde with NADPH by MCR-C fragment, and further reduced to 3HP by MCR-N fragment. In this process, the initial reduction of malonyl-CoA was rate limiting. Site-directed mutagenesis demonstrated that the TGXXXG(A)X(1-2) G and YXXXK motifs were important for enzyme activities of both MCR-N and MCR-C fragments. Moreover, the enzyme activity increased when MCR was separated into two individual fragments. Kinetic analysis showed that MCR-C fragment had higher affinity for malonyl-CoA and 4-time higher K-cat/K-m value than MCR. Dissecting MCR into MCR-N and MCR-C fragments also had a positive effect on the 3HP production in a recombinant Escherichia coli strain. Our study showed the feasibility of protein dissection as a new strategy in biosynthetic systems.
学科主题	生物基化学品
WOS标题词	Science & Technology
类目[WOS]	Multidisciplinary Sciences
研究领域[WOS]	Science & Technology - Other Topics
关键词[WOS]	RECOMBINANT ESCHERICHIA-COLI ; DEHYDROGENASE ; PROTEINS ; PATHWAY ; GENE ; BIOSYNTHESIS ; ACID
收录类别	SCI
语种	英语
WOS记录号	WOS:000324768000081
公开日期	2014-03-24
内容类型	期刊论文
源URL	[http://ir.qibebt.ac.cn:8080/handle/337004/1667]
专题	青岛生物能源与过程研究所_材料生物技术研究中心
作者单位	1.Chinese Acad Sci, Qingdao Inst Bioenergy & Bioproc Technol, Qingdao, Shandong, Peoples R China 2.Chinese Acad Sci, Key Lab Biobased Mat, Qingdao, Shandong, Peoples R China 3.Univ Chinese Acad Sci, Beijing, Peoples R China 4.Chinese Acad Sci, Inst Oceanol, Qingdao, Shandong, Peoples R China
推荐引用方式 GB/T 7714	Liu, Changshui,Wang, Qi,Xian, Mo,et al. Dissection of Malonyl-Coenzyme A Reductase of Chloroflexus aurantiacus Results in Enzyme Activity Improvement[J]. PLOS ONE,2013,8(9):e75554.
APA	Liu, Changshui,Wang, Qi,Xian, Mo,Ding, Yamei,&Zhao, Guang.(2013).Dissection of Malonyl-Coenzyme A Reductase of Chloroflexus aurantiacus Results in Enzyme Activity Improvement.PLOS ONE,8(9),e75554.
MLA	Liu, Changshui,et al."Dissection of Malonyl-Coenzyme A Reductase of Chloroflexus aurantiacus Results in Enzyme Activity Improvement".PLOS ONE 8.9(2013):e75554.