易错PCR技术提高黑曲霉N25植酸酶活力的研究  

Increasing Activity of Phytase from Aspergillus niger N25 by Error-prone PCR

作　　者：冯慧玲[1] 李春梅[1] 吴振芳[1] 陈惠[1] 

机构地区：[1]四川农业大学生命科学与理学院,雅安625014

出　　处：《生物技术通报》2010年第10期226-230,共5页Biotechnology Bulletin

基　　金：国家自然科学基金(30671530)

摘　　要：利用易错PCR技术对黑曲霉(Aspergillus niger)N25的植酸酶基因phyA进行定向进化研究,突变基因产物重组于表达载体pET32a(+)中,并导入大肠杆菌BL21(DE3)构建突变体文库,经筛选获得了最佳突变菌株pET32a-phyAep,其植酸酶活力比出发酶提高了41.8%。突变酶的酶学性质研究发现,与野生酶相比,它的热稳定性,最适温度和最适pH值无显著变化。Random mutagenesis on Aspergillus niger N25 phytase gene phyA was conducted by using error-prune PCR strategy. The error-prone PCR product was recombinated in expression vector pET32a （ ＋ ） , and introducted into Bacillus coli BI21 （DE3） , constructed mutant library. We have obtained the best mutant strain pET32a-phyAep after screening, which has 41.8% higher phytase enzyme activity than original strain. The results showed that the thermostability, the optimum temperature and the optimum pH value did not have the significant change compare with the wild phytase.

关 键 词：植酸酶 定向进化 易错PCR 酶活力 

分 类 号：S816[农业科学—饲料科学] Q816[农业科学—畜牧学]