中文摘要：

为了了解酿酒酵母（Saccharomyces cerevisiae,S.cerevisiae）谷氨酸脱羧酶（GAD）的情况,分别用酵母提取物及胰蛋白胨（3×YP）＋6%棉子糖液体培养基、3×YP＋6%半乳糖液体培养基进行诱导培养,粉碎破碎酵母,采用ProfiniaTM蛋白质纯化系统纯化,并用SDS-PAGE凝胶电泳测定分子质量,同时研究了磷酸吡哆醛（PLP）对酿酒酵母谷氨酸脱羧酶（S.cerevisiaeGAD）活性的影响。结果表明：转基因酿酒酵母在28℃振动培养47h,3×YP＋6%棉子糖培养基OD600值为7.4,3×YP＋6%半乳糖培养基OD600值为6.8。棉子糖诱导的酿酒酵母没有表达目的基因,没有纯化到GAD;相反,半乳糖诱导的酿酒酵母表达了目的基因,纯化到了GAD,其分子质量为67ku。PLP对GAD的活化作用较显著。

英文摘要：

To understand the situation of glutamate decarboxylase （GAD） of Saccharemyces cerevisiae （S. cerevisiae）, 3 x yeast peptone （YP） with 6% raffinose liquid medium and 3 × YP with 6% galactose liquid medium were used for induction culture, respectively. The S. cerevisiae were crushed and broken, and then were purified using ProfiniaTM protein purification system. The molecular weight was determined by SDS - PAGE gel electrophoresis, and the effect of pyridoxal phosphate （PLP） on activity of GAD of S. cerevisiae was studied simultaneously. The re- suits showed that the OD600 value in 3 × YP with 6% ratffinose liquid medium was 7.4 when transgenic S. cerevisiae were used for vibration cul- ture at 28 ℃ for 47 h, and the OD600 value in 3 × YP with 6% galactose liquid medium was 6.8 ; S. cerevisiae induced by raffinose did not express the target gene, and there was no purification for GAD, on the contrary, S. cerevisiae induced by galactose did. The molecular mass was 67 ku, and PLP had obvious activation on GAD.