Combining mutagenesis on Glu281 of prenyltransferase NovQ and metabolic engineering strategies for the increased prenylated activity towards menadione
文献类型：期刊论文
| 作者 | Ni, Wenfeng1,2; Zheng, Zhiming2 ; Liu, Hui2; Wang, Peng2; Wang, Han1,2; Sun, Xiaowen1,2; Yang, Qiang1,2; Fang, Zhiwei1,2; Tang, Hengfang1,2; Zhao, Genhai2
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| 刊名 | APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
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| 出版日期 | 2020-03-03 |
| ISSN号 | 0175-7598 |
| 关键词 | Prenyltransferase NovQ Menadione hydroquinol Site-directed mutagenesis Mevalonate pathway Metabolic engineering |
| DOI | 10.1007/s00253-020-10470-w |
| 通讯作者 | Zheng, Zhiming(zhengzhiming2014@163.com) ; Zhao, Genhai(zhgh327@126.com) |
| 英文摘要 | Prenyltransferase NovQ is a vital class involved in the biosynthesis of secondary metabolites such as clorobiocin and novobiocin. To investigate the relationship between structure and catalytic properties of NovQ, here, we have analyzed the substrate-binding site, namely PT barrel, and revealed that menadione hydroquinol formed intermolecular interactions with the residue Glu281 near the center of the active pocket. In this study, Glu281 was substituted with 9 diverse amino acids and catalytic properties of mutants were observed in vitro. Among them, E281Q showed 2.05-fold activities towards the aromatic substrate and prenyl donor, while others obtained catalytic efficiency between 8.4 and 88.6% of that of wild-type NovQ. Furthermore, the effects of catalytic conditions and substrate status on the activity of NovQ and its mutants were considered to obtain the optimized prenylated reaction. When the evolutionary NovQ variant E281Q was overexpressed in the host constructed to synthesize dimethylallyl diphosphate through the engineered mevalonate (MVA) pathway, we harvested up to 4.7 mg/L prenylated menadione at C-3 position by exogenously supplying the aromatic substrate. The construction of the microbial platform based on NovQ opens a new orientation to further biosynthesize various vitamin K-2 with other ABBA prenyltransferases in E. coli. |
| WOS关键词 | IDENTIFICATION ; CLOQ ; BIOSYNTHESIS ; MENAQUINONES ; EXTRACTION ; MECHANISM ; HOMOLOGS ; RESIDUES ; PLATFORM ; PATHWAY |
| 资助项目 | Key research and development plan of Anhui Province[1804b06020342] ; Natural Science Foundation of Anhui Province[1908085MB48] ; Natural Science Foundation of Anhui Province[1908085MB43] ; China National Key Research and Development Program[2019YFA0904300] ; Major Projects of Science and Technology in Anhui Province[17030801036] |
| WOS研究方向 | Biotechnology & Applied Microbiology |
| 语种 | 英语 |
| 出版者 | SPRINGER |
| WOS记录号 | WOS:000518077800003 |
| 资助机构 | Key research and development plan of Anhui Province ; Natural Science Foundation of Anhui Province ; China National Key Research and Development Program ; Major Projects of Science and Technology in Anhui Province |
| 源URL | [http://ir.hfcas.ac.cn:8080/handle/334002/104003]  |
| 专题 | 中国科学院合肥物质科学研究院 |
| 通讯作者 | Zheng, Zhiming; Zhao, Genhai |
| 作者单位 | 1.Univ Sci & Technol China, Hefei 230026, Anhui, Peoples R China 2.Chinese Acad Sci, Hefei Inst Phys Sci, Key Lab High Magnet Field & Ion Beam Phys Biol, 350 Shushanhu Rd, Hefei 230031, Anhui, Peoples R China |
| 推荐引用方式 GB/T 7714 | Ni, Wenfeng,Zheng, Zhiming,Liu, Hui,et al. Combining mutagenesis on Glu281 of prenyltransferase NovQ and metabolic engineering strategies for the increased prenylated activity towards menadione[J]. APPLIED MICROBIOLOGY AND BIOTECHNOLOGY,2020. |
| APA | Ni, Wenfeng.,Zheng, Zhiming.,Liu, Hui.,Wang, Peng.,Wang, Han.,...&Zhao, Genhai.(2020).Combining mutagenesis on Glu281 of prenyltransferase NovQ and metabolic engineering strategies for the increased prenylated activity towards menadione.APPLIED MICROBIOLOGY AND BIOTECHNOLOGY. |
| MLA | Ni, Wenfeng,et al."Combining mutagenesis on Glu281 of prenyltransferase NovQ and metabolic engineering strategies for the increased prenylated activity towards menadione".APPLIED MICROBIOLOGY AND BIOTECHNOLOGY (2020). |
入库方式： OAI收割
来源：合肥物质科学研究院
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