Cloning and functional identification of a new NADPH-cytochrome P450 reductase in Andrographis paniculata.
10.19540/j.cnki.cjcmm.20171030.011
- Author:
Meng-Die QI
1
;
Jian WANG
2
;
Xiao-Jing MA
2
;
Quan ZHANG
1
;
Fang-Fang WANG
1
;
Ying KANG
1
;
Hui-Xin LIN
2
;
Yong LIU
1
Author Information
1. Beijing University of Chinese Medicine， Beijing 100102， China.
2. 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.
- Publication Type:Journal Article
- Keywords:
andrographolide;
biosynthetic pathway;
cytochrome P450 reductase;
flonicamid;
functional identification;
prokaryotic expression
- MeSH:
Acetates;
Andrographis;
enzymology;
genetics;
Biosynthetic Pathways;
Cloning, Molecular;
Cyclopentanes;
Diterpenes;
metabolism;
NADPH-Ferrihemoprotein Reductase;
genetics;
Oxylipins;
Plant Leaves;
enzymology;
Plant Proteins;
genetics
- From:
China Journal of Chinese Materia Medica
2018;43(2):309-315
- CountryChina
- Language:Chinese
-
Abstract:
Andrographolide is a main active ingredient in traditional Chinese medicine Andrographis paniculata，with a variety of pharmacological activity，widely used in clinical practice. However its biosynthetic pathway has not been resolved. Cytochrome P450 reductase provides electrons for CYP450 and plays an important role in the CYP450 catalytic process. In this study，the coding sequence of A. paniculata CPR was screened and cloned by homologous alignment，named ApCPR4. The ApCPR4 protein was obtained by prokaryotic expression. After isolation and purification，the enzyme activity was identified . The results showed that ApCPR4 could reduce the cytochrome c and ferricyanide in NADPH-dependent manner. In order to verify its function，ApCPR4 and CYP76AH1 were co-transformed into yeast engineering bacteria. The results showed that ApCPR4 could help CYP76AH1 catalyze the formation of rustols in yeast. Real-time quantitative PCR results showed that the expression of ApCPR4 increased gradually in leaves treated with methyl jasmonate (MeJA). The expression pattern was consistent with the trend of induction and accumulation of andrographolide by MeJA，suggesting that ApCPR4 was associated with biosynthesis of andrographolide.
