Heterologous expression of dammarenediol synthase gene in an engineered Saccharomyces cerevisiae.
OBJECTIVE
Dammarenediol production by an engineered yeast Saccharomyces cerevisiae was investigated.


RESULTS
A dammarenediol-producing engineered yeast was constructed by heterologous expression of the dammarenediol synthase gene from Panax ginseng hairy roots through RT-PCR. Fermentation was carried out in a 5-L GRJY-bioreactor with an inoculum size of 1% v/v at 30°C. Dammarenediol detection was performed with silica gel chromatography and HPLC. Determination of dammarenediol synthase activity subcellular distribution was carried out by surveying the enzyme activity in microsomes, lipid particles and total yeast homogenate. When cultured under aerobic conditions, the engineered yeast could produce dammarenediol up to 250μgl(-1). However, when an anaerobic shift strategy was employed, dammarenediol accumulated at a level as twice as that under aerobic condition. The dammarenediol synthase and dammarenediol were mainly localized in lipid particles.


CONCLUSIONS
Dammarenediol could be heterologously produced in engineered yeast. The heterologously expressed dammarenediol synthase is mainly localized in lipid particles. Anaerobic shift strategy could enhance the dammarenediol level in the engineered yeast.


CONCLUSIONS
This study showed that the high-value plant product dammarenediol could be produced by heterologous expression of the according gene in yeast. Furthermore, the anaerobic shift strategy could be potentially applied in oxidosqualene-derived compounds production in yeast. Here, the information about subcellular distribution of heterologously expressed dammarenediol synthase in the engineered yeast was also provided.
Department of Bioengineering, College of Biological and Agricultural Engineering, Jilin University, ChangChun, China.
Y-L
YL
Liang
Department of Bioengineering, College of Biological and Agricultural Engineering, Jilin University, ChangChun, China.
S-J
SJ
Zhao
Department of Bioengineering, College of Biological and Agricultural Engineering, Jilin University, ChangChun, China.
L-X
LX
Xu
Department of Bioengineering, College of Biological and Agricultural Engineering, Jilin University, ChangChun, China.
X-Y
XY
Zhang
D019883:Alkyl and Aryl Transferases / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D019149:Bioreactors; D018076:DNA, Complementary / Q000235:genetics; D005818:Genetic Engineering; D005894:Panax / Q000201:enzymology*; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism*; D012503:Saponins / Q000032:analysis / Q000096:biosynthesis*; D014315:Triterpenes / Q000032:analysis
D018076:DNA, Complementary; D012503:Saponins; D014315:Triterpenes; C106188:dammarenediol; D019883:Alkyl and Aryl Transferases; C514651:dammarenediol-II synthase, Panax ginseng