Production of amorpha-4,11-diene in engineered yeasts.
- Author:
Jian-qiang KONG
1
;
Jun-hao SHEN
;
Yong HUANG
;
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 & Key Laboratory of Bioactive Substances and Resources Utilization of Chinese Herbal Medicine, Ministry of Education of PRC), Beijing 100050, China).
- Publication Type:Journal Article
- MeSH:
Alkyl and Aryl Transferases;
genetics;
metabolism;
DNA, Ribosomal;
genetics;
Fermentation;
Gas Chromatography-Mass Spectrometry;
methods;
Genetic Engineering;
methods;
Genome, Fungal;
genetics;
Plasmids;
Saccharomyces cerevisiae;
genetics;
metabolism;
Sesquiterpenes;
metabolism;
Transformation, Genetic
- From:
Acta Pharmaceutica Sinica
2009;44(11):1297-1303
- CountryChina
- Language:Chinese
-
Abstract:
Plasmid-carrying Saccharomyces cerevisia (W303-1B[pYeDP60/G/ADS]) and genome-transformed S. cerevisia (W303-1B[rDNA:ADS]), both harboring amorpha-4,11-diene synthase (ADS) gene were constructed to investigate the production of amorpha-4,11-diene. The recombinant plasmid pYeDP60/G/ADS that harbors the ADS gene was transformed into S. cerevisiae W303-1B, resulting in the engineered yeast W303-1B[pYeDP60/G/ADS], which contains multi-copies of the plasmid. The ADS gene expression cassette was obtained by PCR amplification of the pYeDP60/G/ADS template, and then introduced into S. cerevisiae W303-1B to obtain the engineered yeast W303-1B[rDNA:ADS], in which the ADS gene was integrated into the rDNA locus of the yeast genome through the homologous recombination. GC-MS analysis confirmed that both of the engineered yeasts could produce amorpha-4,11-diene. Moreover, the amorpha-4,11-diene yield of W303-1B[pYeDP60/G/ADS] was higher than that of W303-1B[rDNA:ADS]. Southern blot analysis showed that there is only one copy of ADS gene in the genome of W303-1B[rDNA:ADS]. It implied that the amorpha-4,11-diene yield can be improved by increasing the ADS gene copies.
