Heterologous expression of dammarenediol synthase gene in an engineered Saccharomyces cerevisiae

被引:12
作者
Liang, Y. -L. [1 ]
Zhao, S. -J. [1 ]
Xu, L. -X. [1 ]
Zhang, X. -Y. [1 ]
机构
[1] Jilin Univ, Coll Biol & Agr Engn, Dept Bioengn, Changchun 130022, Peoples R China
基金
中国国家自然科学基金;
关键词
dammarenediol; hairy roots; heterologous expression; Panax ginseng; Saccharomyces cerevisiae; PANAX-GINSENG; COMBINATORIAL BIOSYNTHESIS; SECONDARY METABOLITES; TOPOLOGY PREDICTION; YEAST; PROTEINS; PROGRESS; ACID;
D O I
10.1111/j.1472-765X.2012.03295.x
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Aims: Dammarenediol production by an engineered yeast Saccharomyces cerevisiae was investigated. Methods and 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 degrees 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 250 mu g l(-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. Significance and Impact of the Study: 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.
引用
收藏
页码:323 / 329
页数:7
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