Construction of Saccharomyces cerevisiae whole-cell biocatalyst system for conversion miltiradiene.
- Author:
Yuan CAI
1
;
Juan GUO
1
;
Yong-Jin ZHOU
2
;
Zhi-Wei ZHU
2
;
Wen-Yan WU
1
;
Lu-Qi HUANG
1
;
Min CHEN
1
;
Zong-Bao ZHAO
2
Author Information
1. National Resource Center for Chinese Material Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China.
2. Department of Biotechnology of Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China.
- Publication Type:Journal Article
- MeSH:
Biosynthetic Pathways;
Biotransformation;
Cytochrome P-450 Enzyme System;
genetics;
metabolism;
Diterpenes;
metabolism;
Diterpenes, Abietane;
biosynthesis;
chemistry;
Electrophoresis, Agar Gel;
Gene Amplification;
NADPH-Ferrihemoprotein Reductase;
genetics;
metabolism;
Open Reading Frames;
Plasmids;
Saccharomyces cerevisiae;
genetics;
metabolism;
Salvia miltiorrhiza;
chemistry
- From:
Acta Pharmaceutica Sinica
2013;48(10):1618-1623
- CountryChina
- Language:Chinese
-
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.
