木糖异构酶在酿酒酵母细胞表面的展示  

Displaying xylose isomerase on yeast cell surface

作　　者：沈煜[1] 侯进[1] 王芳[1] 鲍晓明[1] 曲音波[1] 

机构地区：[1]山东大学微生物技术国家重点实验室,济南250100

出　　处：《工业微生物》2006年第2期12-15,共4页Industrial Microbiology

基　　金：国家自然科学基金资助项目(No.50273019)

摘　　要：将来源于嗜热细菌Thermus thermophilus的木糖异构酶基因xylA,与酿酒酵母(Sac-charomyces cerevisiae)a-凝集素表面展示载体pYD1的Aga2p亚基C端序列融合。编码融合蛋白的基因序列前接上半乳糖诱导型启动子。用LiAc完整细胞法转化酿酒酵母EBY100。含重组质粒的菌株EBY100/pYD-xylA经半乳糖诱导表达外源融合蛋白,免疫荧光显微镜结果显示外源蛋白被锚定在细胞壁上,木糖异构酶活性测定结果表明,细胞壁上酶活测定值为1.52U,木糖异构酶在酿酒酵母细胞壁上得到活性表达。The xylose isomerase （XI） gene from Therrnus thermophilus was fused with the sequence encoding the Cterminal of Aga2p contained in the yeast a- agglutinin surface display vector pYD1. The fused gene was under the control of GAL promoter. The recombinant plasmid was transformed into the yeast stain EBY100 by the lithium acetate method. The strain EBY100/pYD- xyl/A containing the recombinant plasmid was induced by the galactose, and the result of immunofluorescence microscopy confirmed that the fused protein was displayed on the cell wall. The XI activity detected on cell wall was 1.52 U. The results showed that the XI was displayed on the cell wall actively.

关 键 词：木糖异构酶 酿酒酵母 a-凝集素 木糖 

分 类 号：TS261.1[轻工技术与工程—发酵工程]