α-葡萄糖苷酶基因在黑曲霉中的同源表达

Homologous Expression of α-glucosidase Gene in Aspergillus niger

本实验以黑曲霉(Aspergillus niger)内源α-葡萄糖苷酶为基础,对黑曲霉菌株部分序列进行了克隆,构建了能够通过农杆菌介导法整合到黑曲霉基因组中的pSZH-10-6-agd C2表达载体。在转化黑曲霉之后检测到目的基因能够合成蛋白并分泌到细胞外的发酵液中。通过测定发酵液中的重组蛋白对底物的催化能力发现此基因在改造后仍具备酶活性,并且得出此酶的最适反应温度为40℃左右,最适pH值在6左右。

Base on Aspergillus niger endogenous alpha glycosidase enzymes,parts of the Aspergillus niges strain sequences were cloned,the pSZH-10-6-agd C2 expression vector was constructed and integrated into the Aspergillus niger genome by Agrobacterium mediated method.After the transformation of Aspergillus niger,the target gene was detected to be able to synthesize the protein and secreted to extracellular fermented liquid.Through the determination of catalytic ability of recombinant proteins in fermented liquid on substrate,this gene was found still having enzyme activity after this genetic transformation,and it was concluded that the optimal reaction temperature of the enzyme was about 40℃,the optimum pH value was about 6.