糖基转移酶与阿洛酮糖差向异构酶在丝状真菌中高效表达系统的构建

Construction of Glycosyltransferase and Psicose Epimerase Efficient Expression System in Filamentous Fungi

里氏木霉作为一种成熟的丝状真菌工业生产菌株,其拥有优秀的蛋白质分泌能力和真核蛋白质加工修饰过程,因此,越来越多地被作为重组蛋白的生产宿主。本研究构建了里氏木霉胞内表达系统,分别表征了里氏木霉的4个诱导型启动子和9个组成型启动子的表达强度,其中grdh表达强度水平最强。随后,成功利用grdh启动糖基转移酶optEUGT11和阿洛酮糖差向异构酶optBLA表达,且体外均有酶活,表明构建的里氏木霉胞内表达系统表达异源蛋白的可行性。

Trichoderma reesei, a filamentous fungus, is a mature industrial production strain. They have great protein secretion, eukaryotic protein processing and modification abilities. Therefore, they are increasingly used as the production hosts of recombinant proteins. In this study, the intracellular expression system of Trichoderma reesei was constructed,the expression levels of four inducible promoters and nine constitutive promoters of Trichoderma reesei were also characterized. It was found that the expression intensity of grdh was the strongest among them. The grdh promoter was then used to initiate the expression of the glycosyltransferase optEUGT11 and the psicose epimerase optBLA, and these proteins were successfully activated in vitro. The results indicate that the constructed intracellular expression system of Trichoderma reesei can be applied to the expression of the heterologous proteins.