Abstract:

Dammarenediol gene was amplified by RT-PCR from Panax ginseng hairy roots. A engineered saccharomyces cerevisiae was constructed using this gene and the yeast expression vector pAUR123. The positive clone was selected by Aureobasidin A resistance, and was confirmed by Polymerase Chain Reaction (PCR), Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE) and Thin Layer Chromatography (TLC). The results show that the engineered yeast could express functional dammarenediol synthase and produce dammarenediol. Dammarenediol synthase was expressed throughout the growth phase of the engineered yeast. The highest levels of dammarenediol synthase and dammarenediol (132 U/g dried cells and 251 μg/L) in the engineered yeast were observed after 36 h and 42 h growth in liquid YDP medium, respectively.

Key words: bioengineering, dammarenediol synthase, panax ginseng hairy roots, saccharomyces cerevisiae