鲁氏酵母3-磷酸甘油脱氢酶基因（ZrGPD1）在粉状毕赤酵母中的表达

为探索在野生型粉状毕赤酵母（Pichia farinosa）中整合表达来源于耐高渗鲁氏酵母（Zygosacharomyces rouxii）的3-磷酸甘油脱氢酶基因（ZrGPD1）以提高产甘油能力的可行性，应用PCR方法从P. farinosa的染色体中扩增出乳清苷酸脱羧酶基因（URA3）片段，以此作为同源整合的靶序列，构建了整合型表达载体pUR-ZG。电击转化粉状毕赤酵母，以抗生素Zeocin为筛选标记，获得转化子pfa-gu，摇瓶发酵结果表明：以P. farinosa作为对照菌株，发酵72h后，转化子pfa-gu的生物量和甘油含量均高于对照菌株，其中甘油含量达到37g/L，比对照提高了30%。结论：在P. farinosa中异源表达ZrGPD1能够提高细胞的产甘油能力和对渗透压的调节能力。

Heterologous expression of Zygosacharomyces rouxii ZrGpd 1 in Pichia farinosa

To examine the effects of heterologous expression of ZrGPD1 (encoding glycerol 3-phosphate dehydrogenase ) cloned from osmotolerant yeast Zygosacharomyces rouxii on glycerol production in wild Pichia farinosa,the URA3 gene was amplified from P. farinosa as the homology integrative region. A recombinant plasmid (pUR-ZG) was constructed then transformed into P. farinosa by electroporation. The transformant pfa-gu was obtained by the selectable marker Zeocin TM . Primary results showed that the biomass of pfa-gu was higher than the wild type in the flask and after 72h fermentation the concentration of glycerol of pfa-gu was 37g/L enhanced 30% in comparison with the wild type. It is concluded that heterologous expression of ZrGPD1 is useful for increasing glycerol production and the ability of osmoregulation in P. farinosa.