中文摘要：

甘油是一种极其理想的耐高渗透压介质。利用PCR方法，从产甘油假丝酵母W12002-5中扩增出了2个产甘油的关键酶基因GPD和GPP，分别编码3-磷酸甘油脱氢酶（glycerol 3-phosphate dehydrogenase，GPD）和3-磷酸甘油磷酸酶（glycerol 3-phosphate phosphatase，GPP）。利用T-Vector在Escherichia coli JM109中克隆得到大量的GPD和GPP基因，并成功构建了重组质粒pYX212-GPD和pYX212．GPP；通过LiAc转化法将重组质粒导入酿酒酵母Saccharomyces cerevisiae W303-1A。初步实验结果表明：发酵过程中pYX212-GPD／S．cerevisiae W303-1A的生物量高于pYX212-GPP／S．cerevisiae W303-1A和野生型S．cerevisiae W303-1A；发酵72h后，pYX212-GPD／S．cerevisiae W303-1A发酵液中甘油含量大约为12mmol／L，明显高于野生型S．cerevisiaeW303-1A的甘油含量，而pYX212-GPP／S．cerevisiae W303-1A与野生型S．cerevisiae W303-1A在甘油含量上相差不大，均只有4mmol／L左右。

英文摘要：

Glycerol is an ideal osmotolerant medium. Two genes of GPD and GPP encoding glycerol 3- phosphate dehydrogenase and glycerol 3-phosphate phosphatase, key enzymes in glycerol synthesis, were cloned from Candida glycerolgenesis WL2002-5 using PCR method. In Escherichia coli JM109, enough GPD and GPP genes were amplified using T-vector, furthermore, expression plasmids pYX212-GPD containing GPD gene and pYX212-GPP containing GPP gene were constructed, which were introduced into Saccharomyces cerevisiae W303- 1A using LiAc transformation method successfully. Primary results showed that the biomass of pYX212-GPD/S. cerevisiae W303-1A was higher than those of pYX212-GPP/S, cerevisiae W303-1A and the wild type during the fermentation. After 72h fermentation, introduction of GPD gene could increase glycerol production to about 12mmoL/L, while GPP gene had little effect on glycerol production （only 4mmoL/L） in comparison with the wild type.