摘要
为探索在野生型粉状毕赤酵母(Pichiafarinosa)中整合表达来源于耐高渗鲁氏酵母(Zygosacharomycesrouxii)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%。因此,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。
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 ZeocinTM. 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.
出处
《中国生物工程杂志》
CAS
CSCD
北大核心
2006年第8期32-36,共5页
China Biotechnology
关键词
甘油
3-磷酸甘油脱氢酶
耐高渗鲁氏酵母
整合表达
Glycerol Glycerol 3-phosphate dehydrogenase Osmotolerant yeast Zygosaccharomyces rouxii Integrative expression