Directed evolution of mevalonate kinase in Escherichia coli by random mutagenesis for improved lycopene
文献类型：期刊论文
| 作者 | Chen, Hailin1,2; Liu, Changqing1; Li, Meijie1; Zhang, Haibo1 ; Xian, Mo1; Liu, Huizhou1
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| 刊名 | RSC ADVANCES
 |
| 出版日期 | 2018 |
| 卷号 | 8期号:27页码:15021-15028 |
| ISSN号 | 2046-2069 |
| DOI | 10.1039/c8ra01783b |
| 文献子类 | Article |
| 英文摘要 | 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. |
| WOS关键词 | ISOPRENOID BIOSYNTHETIC-PATHWAY ; HMG-COA REDUCTASE ; SACCHAROMYCES-CEREVISIAE ; BETA-CAROTENE ; METHANOCOCCUS-JANNASCHII ; ISOPENTENYL DIPHOSPHATE ; MOLECULAR-CLONING ; IN-VITRO ; GENE ; INHIBITION |
| WOS研究方向 | Chemistry |
| 语种 | 英语 |
| WOS记录号 | WOS:000430801400036 |
| 出版者 | ROYAL SOC CHEMISTRY |
| 资助机构 | 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) |
| 源URL | [http://ir.qibebt.ac.cn/handle/337004/11493]  |
| 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. |
入库方式： OAI收割
来源：青岛生物能源与过程研究所
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