Overexpression and isolation of CYP76AH1 in Salvia miltiorrhiza hairy roots.
10.19540/j.cnki.cjcmm.2017.0157
- Author:
Meng-Die QI
1
;
Jian WANG
1
;
Xiao-Jing MA
1
;
Quan ZHANG
2
;
Fang-Fang WANG
2
;
Ying KANG
2
;
Yong LIU
2
;
Hui-Xin LIN
1
Author Information
1. State Key Laboratory Breeding Base of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China.
2. Beijing University of Chinese Medicine, Beijing 100102, China.
- Publication Type:Journal Article
- Keywords:
CYP76AH1;
detergent;
protein complexes;
secondary metabolism;
tanshinone;
transgenic hairy root
- From:
China Journal of Chinese Materia Medica
2017;42(20):3901-3905
- CountryChina
- Language:Chinese
-
Abstract:
Protein complexes are involved in the synthesis of multiple secondary metabolites in plants, and their separation is essential to elucidate plant secondary metabolism and improve in vitro catalytic efficiency. In this study, the transgenic hairy roots of CYP76AH1, a key enzyme of tanshinone synthesis pathway, was constructed and the transgenic hairy roots of Danshen overexpressing CYP76AH1 protein were screened by Western blotting and used as a tissue culture material for the subsequent extraction of protein complex in tanshinone synthesis pathway. By optimizing the type and concentration of the detergent in the protein extraction buffer, the buffer containing 0.5% Triton X-100 was selected as the best extraction buffer, and a relatively large amount of soluble CYP76AH1 protein was isolated. This study lays the foundation for the further separation and purification of protein complexes interacting with CYP76AH1, and provides the idea for deep analysis of tanshinone metabolic pathway.
