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学科主题: 物理化学
题名: CYP76AH1 catalyzes turnover of miltiradiene in tanshinones biosynthesis and enables heterologous production of ferruginol in yeasts
作者: Guo, Juan1;  Zhou, Yongjin J.2;  Hillwigc, Matthew L.3;  Shen, Ye1;  Yang, Lei4, 5;  Wang, Yajun1;  Zhang, Xianan1;  Liu, Wujun2;  Peters, Reuben J.3;  Chen, Xiaoya4, 5;  Zhao, Zongbao K.2;  Huang, Luqi1
通讯作者: 赵宗保 ;  Peters RJ ;  黄璐琦
关键词: phytoterpenoids biosynthesis ;  gene discovery ;  synthetic pathway ;  metabolic engineering
刊名: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
发表日期: 2013-07-16
DOI: 10.1073/pnas.1218061110
卷: 110, 期:29, 页:12108-12113
收录类别: SCI
合作性质: 
文章类型: Article
部门归属: 18
项目归属: 1816
产权排名: 待补充
WOS标题词: Science & Technology
类目[WOS]: Multidisciplinary Sciences
资助者: 2,2
研究领域[WOS]: Science & Technology - Other Topics
英文摘要: Cytochrome P450 enzymes (CYPs) play major roles in generating highly functionalized terpenoids, but identifying the exact biotransformation step(s) catalyzed by plant CYP in terpenoid biosynthesis is extremely challenging. Tanshinones are abietane-type norditerpenoid naphthoquinones that are the main lipophilic bioactive components of the Chinese medicinal herb danshen (Salvia miltiorrhiza). Whereas the diterpene synthases responsible for the conversion of (E,E,E)-geranylgeranyl diphosphate into the abietane miltiradiene, a potential precursor to tanshinones, have been recently described, molecular characterization of further transformation of miltiradiene remains unavailable. Here we report stable-isotope labeling results that demonstrate the intermediacy of miltiradiene in tanshinone biosynthesis. We further use a next-generation sequencing approach to identify six candidate CYP genes being coregulated with the diterpene synthase genes in both the rhizome and danshen hairy roots, and demonstrate that one of these, CYP76AH1, catalyzes a unique four-electron oxidation cascade on miltiradiene to produce ferruginol both in vitro and in vivo. We then build upon the previous establishment of miltiradiene production in Saccharomyces cerevisiae, with incorporation of CYP76AH1 and phyto-CYP reductase genes leading to heterologous production of ferruginol at 10.5 mg/L. As ferruginol has been found in many plants including danshen, the results and the approaches that were described here provide a solid foundation to further elucidate the biosynthesis of tanshinones and related diterpenoids. Moreover, these results should facilitate the construction of microbial cell factories for the production of phytoterpenoids.
关键词[WOS]: GENE-CLUSTER ;  SALVIA-MILTIORRHIZA ;  TERPENOID BIOSYNTHESIS ;  RICE ;  PATHWAY ;  CYTOCHROME-P450 ;  IDENTIFICATION ;  DITERPENES ;  DIVERSIFICATION ;  FOSMIDOMYCIN
语种: 英语
原文出处: 查看原文
WOS记录号: WOS:000322086100093
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/119582
Appears in Collections:中国科学院大连化学物理研究所_期刊论文

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作者单位: 1.China Acad Chinese Med Sci, Nat Resource Ctr Chinese Mat Med, Beijing 100700, Peoples R China
2.Chinese Acad Sci, Dalian Inst Chem Phys, Div Biotechnol, Dalian 116023, Peoples R China
3.Iowa State Univ, Dept Biochem Biophys & Mol Biol, Ames, IA 50011 USA
4.Chinese Acad Sci, Shanghai Inst Biol Sci, Natl Key Lab Plant Mol Genet, Inst Plant Physiol & Ecol, Shanghai 200032, Peoples R China
5.Chinese Acad Sci, Shanghai Inst Biol Sci, Plant Sci Res Ctr, Shanghai Chenshan Bot Garden, Shanghai 200032, Peoples R China

Recommended Citation:
Guo, Juan,Zhou, Yongjin J.,Hillwigc, Matthew L.,et al. CYP76AH1 catalyzes turnover of miltiradiene in tanshinones biosynthesis and enables heterologous production of ferruginol in yeasts[J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA,2013,110(29):12108-12113.
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