Engineering a D-lactate dehydrogenase that can super-efficiently utilize NADPH and NADH as cofactors

CORC > 天津工业生物技术研究所 > 生物催化与合成生物学研究祝俊/林建平 > 期刊论文

	Engineering a D-lactate dehydrogenase that can super-efficiently utilize NADPH and NADH as cofactors
	Meng, Hengkai 1,2; Liu, Pi 3; Sun, Hongbing 2,3; Cai, Zhen 2; Zhou, Jie 2; Lin, Jianping 3,4; Li, Yin 2
刊名	SCIENTIFIC REPORTS
	2016-04-25
卷号	6
英文摘要	Engineering the cofactor specificity of a natural enzyme often results in a significant decrease in its activity on original cofactor. Here we report that a NADH-dependent dehydrogenase (D-LDH) from Lactobacillus delbrueckii 11842 can be rationally engineered to efficiently use both NADH and NADPH as cofactors. Point mutations on three amino acids (D176S, I177R, F178T) predicted by computational analysis resulted in a modified enzyme designated as D-LDH. The K-cat/K-m of the purified D-LDH on NADPH increased approximately 184-fold while the K-cat/K-m on NADH also significantly increased, showing for the first time that a rationally engineered D-LDH could exhibit comparable activity on both NADPH and NADH. Further kinetic analysis revealed that the enhanced affinity with NADH or NADPH and the significant increased K-cat of D-LDH* resulted in the significant increase of S-LDH* activity on both NADPH and NADH. This study thus demonstrated that the cofactor specificity of dehydrogenase can be broadened by using targeted engineering approach, and the engineered enzyme can efficiently function in NADH-rich, or NADPH-rich, or NADH and NADPH-rich environment.
WOS标题词	Science & Technology
类目[WOS]	Multidisciplinary Sciences
研究领域[WOS]	Science & Technology - Other Topics
关键词[WOS]	SYNECHOCYSTIS SP PCC6803 ; SP PCC 6803 ; LACTIC-ACID ; PHOTOSYNTHETIC PRODUCTION ; LACTOBACILLUS-BULGARICUS ; SUBSTRATE-SPECIFICITY ; COENZYME SPECIFICITY ; CYANOBACTERIUM ; REDUCTASE ; EVOLUTION
收录类别	SCI
语种	英语
WOS记录号	WOS:000374655100001
内容类型	期刊论文
源URL	[http://124.16.173.210/handle/834782/2255]
专题	天津工业生物技术研究所_生物催化与合成生物学研究祝俊/林建平_期刊论文
作者单位	1.Univ Sci & Technol China, Dept Cellular Biol, Hefei 230026, Peoples R China 2.Chinese Acad Sci, Inst Microbiol, CAS Key Lab Microbial Physiol & Metab Engn, Beijing, Peoples R China 3.Chinese Acad Sci, Tianjin Inst Ind Biotechnol, Tianjin, Peoples R China 4.Nankai Univ, Coll Pharm, Tianjin 300071, Peoples R China
推荐引用方式 GB/T 7714	Meng, Hengkai,Liu, Pi,Sun, Hongbing,et al. Engineering a D-lactate dehydrogenase that can super-efficiently utilize NADPH and NADH as cofactors[J]. SCIENTIFIC REPORTS,2016,6.
APA	Meng, Hengkai.,Liu, Pi.,Sun, Hongbing.,Cai, Zhen.,Zhou, Jie.,...&Li, Yin.(2016).Engineering a D-lactate dehydrogenase that can super-efficiently utilize NADPH and NADH as cofactors.SCIENTIFIC REPORTS,6.
MLA	Meng, Hengkai,et al."Engineering a D-lactate dehydrogenase that can super-efficiently utilize NADPH and NADH as cofactors".SCIENTIFIC REPORTS 6(2016).