Recent advances in the study of amorpha-4,11-diene synthase and its metabolic engineering.
- Author:
Jian-Qiang KONG
1
;
Yong HUANG
;
Jun-Hao SHEN
;
Wei WANG
;
Ke-Di CHENG
;
Ping ZHU
Author Information
1. Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Key Laboratory of Biosynthesis of Natural Products, Ministry of Health of PRC, Beijing 100050, China.
- Publication Type:Journal Article
- MeSH:
Alkyl and Aryl Transferases;
biosynthesis;
genetics;
Amino Acid Sequence;
Antimalarials;
metabolism;
Arabidopsis;
enzymology;
genetics;
Artemisia annua;
enzymology;
genetics;
Artemisinins;
metabolism;
Aspergillus;
genetics;
metabolism;
Cloning, Molecular;
DNA, Complementary;
genetics;
Escherichia coli;
genetics;
metabolism;
Metabolic Engineering;
Saccharomyces cerevisiae;
genetics;
metabolism;
Tobacco;
enzymology;
genetics
- From:
Acta Pharmaceutica Sinica
2009;44(12):1320-1327
- CountryChina
- Language:Chinese
-
Abstract:
Amorpha-4,11-diene synthase (ADS) can convert farnesyl pyrophosphate (FPP) to amorpha-4, 11-diene, a precursor of artemisinin. ADS plays an important role in the biosynthesis of artemisinin. This review summarizes the molecular biology and metabolic engineering study of ADS in recent years. The genomic DNA and its cDNA sequences of amorpha-4, 11-diene synthase were cloned from Artemisia annua L. The cDNA encoding amorpha-4, 11-diene synthase contains a 1 641 bp open reading frame coding for 546 amino acids. ADS shows a broad pH optimum and an absolute requirement for divalent metal ions as cofactors. The specificity of ADS to the substrates and products is not high and the formation of amorpha-4, 11-diene by ADS from FPP is achieved by an initial 1, 6-closure with subsequent 1, 10-closure. The ADS cDNA cloned from Artemisia annua L, or totally synthesized by PCR, was introduced into different hosts including E. coli, S. cerevisiae, Nicotiana tabacum L. Arabidopsis thaliana and A. nidulans resulting in varied engineering microorganisms and cells producing amorpha-4, 11-diene. The way to improve the production of amorpha-4, 11-diene was investigated by two strategies such as improving the supply of substrate and directing FPP flux to amorpha-4, 11-diene production from competing pathways.
