title: [Synergistic effect of amorpha-4,11-diene synthase gene in engineered Saccharomyces cerevisiae].
abstract: To construct an engineered Saccharomyces cerevisiae producing high titres of amorpha-4,11-diene, we investigated the possible synergistic effect of different vectors containing amorpha-4,11-diene synthase(ADS) gene within one yeast cell. We constructed the ADS recombinant plasmid pGADADS. This plasmid and another ADS recombinant plasmid pYeDP60/G/ADS were alone, or co-transformed into yeast Saccharomyces cerevisiae W303-1B and WK1, respectively, resulting in the following engineered yeasts, W303B[pGADADS], W303B[pYGADS], W303B[pYGADS+pGADADS], WK1[pGADADS], WK1[pYGADS] and WK1[pYGADS+pGADADS]. All of the six strains were cultured for GC-MS analysis of amorpha-4,11-diene. The results showed that all of the engineered yeasts could produce amorpha-4,11-diene. The yield of the product was improved with increasing ADS gene copies while no deleterious effect on the strain growth was found. Moreover, the product yield of the engineered yeast co-transformed with multiple plasmids was much higher than the total yield of the different engineered yeasts with only one plasmid, respectively. In conclusion, there was a distinct synergistic effect between different recombinant ADS plasmids within one cell. Our results facilitate the construction of the engineered yeast with high yield of amorpha-4,11-diene, the precursor of artemisinin.
authors.affiliation: 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, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, China. jianqiangk@imm.ac.cn
authors.forename: Jianqiang
authors.initials: J
authors.lastname: Kong
authors.forename: Xiaohui
authors.initials: X
authors.lastname: Zhi
authors.forename: Wei
authors.initials: W
authors.lastname: Wang
authors.forename: Kedi
authors.initials: K
authors.lastname: Cheng
authors.forename: Ping
authors.initials: P
authors.lastname: Zhu
mesh_terms: D019883:Alkyl and Aryl Transferases / Q000096:biosynthesis / Q000235:genetics*; D037621:Artemisinins / Q000737:chemistry / Q000378:metabolism*; D005818:Genetic Engineering / Q000379:methods; D005822:Genetic Vectors / Q000235:genetics*; D011995:Recombination, Genetic; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*
chemical_list: D037621:Artemisinins; D019883:Alkyl and Aryl Transferases; C403714:amorpha-4,11-diene synthase