title: [Production of amorpha-4,11-diene in engineered yeasts].
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.
authors.affiliation: 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).
authors.forename: Jian-qiang
authors.initials: JQ
authors.lastname: Kong
authors.forename: Jun-hao
authors.initials: JH
authors.lastname: Shen
authors.forename: Yong
authors.initials: Y
authors.lastname: Huang
authors.forename: Wei
authors.initials: W
authors.lastname: Wang
authors.forename: Ke-di
authors.initials: KD
authors.lastname: Cheng
authors.forename: Ping
authors.initials: P
authors.lastname: Zhu
mesh_terms: D019883:Alkyl and Aryl Transferases / Q000235:genetics* / Q000378:metabolism; D004275:DNA, Ribosomal / Q000235:genetics; D005285:Fermentation; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods; D005818:Genetic Engineering / Q000379:methods*; D016681:Genome, Fungal / Q000235:genetics; D010957:Plasmids; D000081223:Polycyclic Sesquiterpenes; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D012717:Sesquiterpenes / Q000378:metabolism*; D014170:Transformation, Genetic
chemical_list: D004275:DNA, Ribosomal; D000081223:Polycyclic Sesquiterpenes; D012717:Sesquiterpenes; C515348:amorpha-4,11-diene; D019883:Alkyl and Aryl Transferases; C403714:amorpha-4,11-diene synthase