title: [Construction of Saccharomyces cerevisiae whole-cell biocatalyst system for conversion miltiradiene].
abstract: Tanshinones are the bioactive components of the Chinese medicinal herb Salvia miltiorrhiza, while its biosynthetic pathway remains to be characterized. Rapid identification and characterization of the genes correlated to tanshinones biosynthesis is very important. As one of the intermediates of tanshinones biosynthesis, the ferruginol content is relative low in both root and engineered bacteria. It is urgent to construct an efficient system for conversion of miltiradiene to ferruginol to obtain large amount of ferruginol as the substrates for further identifying other downstream genes involved in tanshinones biosynthesis. In this study, we constructed the whole-cell yeast biocatalysts co-expressing miltiradiene oxidase CYP76AH1 and cytochrome P450 reductases (SmCPR1) from Salvia miltiorrhiza, and then characterized it with RT-PCR. After permeabilization, the yeast whole-cell could catalyze turnover of miltiradiene to ferruginol efficiently through single-step biotransformation with a conversion efficiency up to 69.9%. The yeast whole-cell biocatalyst described here not only provide an efficient platform for producing ferruginol in recombinant yeast but also an alternative strategy for identifying other CYP genes involved in tanshinones biosynthesis.
authors.affiliation: National Resource Center for Chinese Material Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Cai
authors.affiliation: National Resource Center for Chinese Material Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China.
authors.forename: Juan
authors.initials: J
authors.lastname: Guo
authors.affiliation: Department of Biotechnology of Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China.
authors.forename: Yong-Jin
authors.initials: YJ
authors.lastname: Zhou
authors.affiliation: Department of Biotechnology of Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China.
authors.forename: Zhi-Wei
authors.initials: ZW
authors.lastname: Zhu
authors.affiliation: National Resource Center for Chinese Material Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China.
authors.forename: Wen-Yan
authors.initials: WY
authors.lastname: Wu
authors.affiliation: National Resource Center for Chinese Material Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China.
authors.forename: Lu-Qi
authors.initials: LQ
authors.lastname: Huang
authors.affiliation: National Resource Center for Chinese Material Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China.
authors.forename: Min
authors.initials: M
authors.lastname: Chen
authors.affiliation: Department of Biotechnology of Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China.
authors.forename: Zong-Bao
authors.initials: ZB
authors.lastname: Zhao
mesh_terms: D045784:Abietanes / Q000096:biosynthesis* / Q000737:chemistry; D053898:Biosynthetic Pathways; D001711:Biotransformation; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics / Q000378:metabolism; D004224:Diterpenes / Q000378:metabolism*; D004587:Electrophoresis, Agar Gel; D005784:Gene Amplification; D009251:NADPH-Ferrihemoprotein Reductase / Q000235:genetics / Q000378:metabolism; D016366:Open Reading Frames; D010957:Plasmids; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D027561:Salvia miltiorrhiza / Q000737:chemistry*
chemical_list: D045784:Abietanes; D004224:Diterpenes; C431074:ferruginol; C573600:miltiradiene; C021751:tanshinone; D003577:Cytochrome P-450 Enzyme System; D009251:NADPH-Ferrihemoprotein Reductase