摘要
为构建具有分泌黑曲霉β-葡萄糖苷酶活力的酵母工程菌,探讨β-葡萄糖苷酶基因(bgl基因)在酵母菌中的表达情况.克隆了黑曲霉bgl基因的cDNA片段,利用质粒pPICZαA构建了该基因的酵母表达载体,经线性化后采用电穿孔法转入毕赤酵母菌中.以甲醇为诱导物进行诱导培养,检测结果表明黑曲霉bgl基因已成功转入酵母菌中并可正常表达,重组蛋白具有β-葡萄糖苷酶水解活性,在诱导培养72 h后,发酵液的酶活力可达到0.78 U/mL.
In order to construct an engineering yeast strain capable of secreting β-1,4-glucosidase of Aspergillus niger,the article studied the expression of β-1,4-glucosidase gene（bgl） in yeast.The cDNA of bgl gene of Aspergillus niger was cloned,and a yeast expression vector of the bgl gene cDNA was constructed by using pPICZαA plasmid.The vector was linearized and transferred into Pichia pastoris by electroporation method.The results showed that the gene was transferred to Pichia pastoris and was expressed successfully through induction culture in methanol as the inducer;the recombinant protein had the activity in hydrolyzing β-1,4-glucosidase;after induction culture for 72 hours,the enzyme activity of the borth was up to 0.78 U/mL.
出处
《河南工业大学学报（自然科学版）》
CAS
北大核心
2010年第5期42-45,共4页
Journal of Henan University of Technology：Natural Science Edition
基金
河南工业大学校博士科研启动基金项目
河南省优秀青年骨干教师项目(155)
关键词
黑曲霉
毕赤酵母
Β-葡萄糖苷酶基因
表达
Aspergillus niger
Pichia Pastoris
β-1,4-glucosidase gene
expression
