纤维素酶嵌合体EGX-Gluc1C的构建与表达

Construction and Expression of the Chimera Cellulase EGX-Gluc1C

[目的]设计和构建一种由β-糖苷水解酶Gluc1C和多功能纤维素酶EGX融合而成的纤维素酶嵌合体EGX-Gluc1C。[方法]首先构建了嵌合体的质粒,采用GST标准纯化方法,分别获得重组蛋白Gluc1C、EGX、纤维素酶嵌合体EGX-Gluc1C,并且制备了抗Gluc1C的血清。[结果]通过GST纯化的3个蛋白纯度达90%,可以进行后续的酶活检测;通过Western blot验证,发现抗Gluc1C的血清效价很高(稀释比例达1∶5 000)。[结论]该研究可为纤维素酶的工业化应用提供一定理论依据。

[Objective]To design and construct chimera cellulase EGX-GluclC originated from the fusion of(3-glucosidase GluclC and a cullulase EGX.[Method]First,we generated the plasmid inserting chimera cellulase.Then,we purified the recombinant GluclC,EGX and EGXGluclC according to the standard GST purification manual.We also generated the anti-GluclC serum from the rabbit.[Result]The coomassie staining results demonstrated that the purity of these proteins was90%,which was sufficient for enzyme catalytic assay.Through verification of western blot,the anti-GluclC serum produced from rabbit was of high quality and the dilution ratio can be1:5000.[Conclusion]The study provided a theoretical basis for the industrial application of cellulase.