摘要
从瑞氏木霉(Trichoderma reesei)cDNA文库中克隆到外切葡聚糖纤维二糖水解酶I(CBHI)cDNA基因,将该基因分别连接到酿酒酵母(Saccharom yces cerevisiae)表达载体pAJ401和pV T100_U上,构建了重组质粒pA J401-cbh1和pV T100_U-cbh1。分别电转化2个重组质粒转移至酿酒酵母H158中,得到重组酵母HPC和HAC,实现了CBHI的分泌型表达。再将质粒pA J401-cbh1转入已含有瑞氏木霉内切葡聚糖酶I基因eg1的重组酵母H 1m中,构建了同时表达cbh1和eg1的重组酵母菌株HMEPC。比较HMEPC和H1m对纤维素底物的降解,前者滤纸酶活比后者提高14.3%,表明CBHI与EGI对滤纸降解有协同作用。
The cbhl gene was obtained by PCR from T.reesei cDNA library and was inserted into S.cerevisiae expression vector pAJ401 and pVT 100_U to construct the expression cassette of cbh 1. The cbh 1 gene was transferred to S.cerevisiae on the different expression plasmid, and CBHI was produced by recombinant S. cerevisiae strains HPC and HAC with cellobiohydrolases activities 4 U/L and 3.5 U/L respectively. CBHI and EGI co-expressing recombinant S.cerevisiae strain HMEPC was constructed. The filter paper activity of HMEPC increased 14.3 % than that ofHlm, which indicated the synergism between cbhl and egl. （Tran. by YUE Yang）
出处
《酿酒科技》
北大核心
2005年第9期28-30,35,共4页
Liquor-Making Science & Technology
关键词
瑞氏木霉
纤维素酶基因
酿酒酵母
Trichoderrna reesei
cellobiohydrolases Ⅰ
gene expression
Saccharomyces cerevisiae
