Directed evolution of mevalonate kinase in Escherichia coli by random mutagenesis for improved lycopene

doi:10.1039/c8ra01783b

QIBEBT-IR

	Directed evolution of mevalonate kinase in Escherichia coli by random mutagenesis for improved lycopene
	Chen, Hailin 1,2; Liu, Changqing 1; Li, Meijie 1; Zhang, Haibo1 ; Xian, Mo1 ; Liu, Huizhou1
	2018
发表期刊	RSC ADVANCES
ISSN	2046-2069
卷号	8 期号:27 页码:15021-15028
摘要	Lycopene is a terpenoid pigment that has diverse applications in the fields of food and medicine. Metabolic engineering in microbial hosts has shown that mevalonate kinase (MK, EC2.7.1.366) is one of the rate-limiting enzymes in the lycopene synthetic pathway. In this study, a directed evolution strategy in Escherichia coli was used to optimize the activity of Saccharomyces cerevisiae MK. Using three rounds of error-prone PCR; screening the development of a lycopene-dependent color reaction; and combinatorial site-specific saturation mutagenesis, three activity-enhancing mutations were identified: V13D, S148I, and V301E. V13D was near the MK catalytic center, in the -sheet that forms a salt-bridge with nearby Arg-248. S148I was located in the -helix lid and improved the stability of the -helix. V301E may increase MK folding by influencing its secondary structure. The Km (RS)-mevalonate of purified mutant MK decreased by 74% compared with the Km (RS)-mevalonate of the wild-type MK, and the Kcat (RS)-mevalonate was improved by 26% compared with wild type. Fermentation experiments revealed that lycopene production of the mutant MK increased 2.4-fold compared with wild-type MK.
文章类型	Article
WOS标题词	Science & Technology ; Physical Sciences
DOI	10.1039/c8ra01783b
关键词[WOS]	ISOPRENOID BIOSYNTHETIC-PATHWAY ; HMG-COA REDUCTASE ; SACCHAROMYCES-CEREVISIAE ; BETA-CAROTENE ; METHANOCOCCUS-JANNASCHII ; ISOPENTENYL DIPHOSPHATE ; MOLECULAR-CLONING ; IN-VITRO ; GENE ; INHIBITION
收录类别	SCI
语种	英语
WOS研究方向	Chemistry
项目资助者	National Natural Science Foundation of China(21572242) ; Taishan Scholars Climbing Program of Shandong(tspd20150210) ; National Natural Science Foundation of China (NSF)(31400084) ; Youth Innovation Promotion Association CAS(2017252)
WOS类目	Chemistry, Multidisciplinary
WOS记录号	WOS:000430801400036
出版者	ROYAL SOC CHEMISTRY
引用统计
文献类型	期刊论文
条目标识符	http://ir.qibebt.ac.cn/handle/337004/11494
专题	中国科学院青岛生物能源与过程研究所
通讯作者	Zhang, Haibo; Xian, Mo
作者单位	1.Chinese Acad Sci, Qingdao Inst Bioenergy & Bioproc Technol, CAS Key Lab Biobased Mat, 189 Songling Rd, Qingdao 266101, Peoples R China 2.Univ Chinese Acad Sci, Sino Danish Coll, 19 A Yuquan Rd, Beijing 100049, Peoples R China
推荐引用方式 GB/T 7714	Chen, Hailin,Liu, Changqing,Li, Meijie,et al. Directed evolution of mevalonate kinase in Escherichia coli by random mutagenesis for improved lycopene[J]. RSC ADVANCES,2018,8(27):15021-15028.
APA	Chen, Hailin,Liu, Changqing,Li, Meijie,Zhang, Haibo,Xian, Mo,&Liu, Huizhou.(2018).Directed evolution of mevalonate kinase in Escherichia coli by random mutagenesis for improved lycopene.RSC ADVANCES,8(27),15021-15028.
MLA	Chen, Hailin,et al."Directed evolution of mevalonate kinase in Escherichia coli by random mutagenesis for improved lycopene".RSC ADVANCES 8.27(2018):15021-15028.