宇佐美曲霉木聚糖酶基因xynⅡ在不同毕赤酵母中的分泌表达  

Secreted expression of Aspergillus usamii xylanase gene in different Pichia pastoris

作　　者：周晨妍[1] 邬敏辰[2] 李东峰[1] 王瑾[1] 王武[1] 

机构地区：[1]江南大学工业生物技术教育部重点实验室,无锡214122 [2]江南大学医药学院,无锡214122

出　　处：《生物加工过程》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)

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...

关 键 词：宇佐美曲霉 内切-1 4-木聚糖酶基因 毕赤酵母 分泌表达 

分 类 号：Q78[生物学—分子生物学]