摘要
将宇佐美曲霉E001的内切-1,4-木聚糖酶基因克隆到毕赤酵母表达载体pPIC9K中,得到重组质粒pPXY-NII,将其经SalⅠ线性化后分别转化2株毕赤酵母GS115和KM71,xynⅡ基因通过同源重组被整合到毕赤酵母染色体上,并处于酵母α因子的下游,经筛选获得阳性重组菌PXGL98(Mut+)和PXKL29(Muts)。该木聚糖酶基因在2株毕赤酵母中均实现了分泌表达。同时对工程菌的发酵条件进行了优化,在甲醇诱导下,PXGL98与PXKL29培养物上清液中的酶活力分别可达1156.92 U/mL和1646.03 U/mL。
The endo-1,4-xylanase gene from Aspergillus usamii E001 was cloned into the Pichia pastoris expression vector,pPIC9K,the recombinant plasmid was named pPXYNII.The recombinant plasmid pPXYNII was linearized with SalⅠand then transformed into Pichia pastoris GS115 and KM71,respectively.The xynⅡgene was in frame integrated into the Pichia genome through homologous recombination.The recombinant Pichia pastoris GS115(PXGL98) and KM71(PXKL29) secreted functional endo-1,4-xylanase,and the enzymatic activities reac...
出处
《生物加工过程》
CAS
CSCD
2008年第2期38-42,共5页
Chinese Journal of Bioprocess Engineering
基金
国家863资助项目(2006AA10Z305)
