Separation and Purification of GST-glycerol-3-phosphate Dehydrogenase

作     者：Hongmei ZHAO Shihai LI Yasuo WATANABE 

作者机构：College of Animal SciencesYangtze University Agriculture DepartmentEhime University 

出 版 物：《Agricultural Biotechnology》 (农业生物技术（英文版）)

年 卷 期：2016年第5卷第5期

页      面：44-45页

学科分类：0710[理学-生物学] 071010[理学-生物化学与分子生物学] 07[理学] 

基　　金：Supported by Social Service Project of New Countryside Development Research Institute of Yangtze University(201411) 

主　　题：Saccharomyces cerevisiae Glycerol-3-phosphate dehydrogenase Galactose SDS-PAGE gel electrophoresis Separation and purification 

摘      要：In order to investigate the expression of glycerol-3 -phosphate dehydrogenase by GCY1 gene in recombinant Saccharomyces cerevisiae, induction culture of the S. cerevisiaestrain was performed with SD-URA 2% galactose, 3 × YP ＋ 6% glucose, SC-URA 2% galactose, and SC-URA 2% galactose ＋ 5% NaCI glyeerol-3-phosphate dehydregenase, the cultured S. cerevisiaewas comminuted followed by full-automatic high-speed purification, and SDS-PAGE gel electrophoresis was performed for molecular weight of the GST fusion protein. The results showed that after shaking culture of the S. cerevisiae containing GCY1 at 25 ℃, the OD values of its 3 × YP ＋ 6% glucose culture and SC-URA 2% galaetose ＋ 5% NaC1 culture were 8.75 and 7.35, respectively. It was shown by purification with a Profinia low-pressure liquid chromatograph that only the S. cerevisiae cultured in SC-URA 2% galactose ＋ 5% NaC1 medium expressed glycerel-3-phosphate de- hydrogenase, the molecular weight of which was detected as 65 ku by SDS-PAGE gel electrophoresis.