甘薯链霉菌中潜在纤维素酶增效蛋白基因克隆、表达及产物活性检测

Cloning,Expression and Activity Detection of Potential Cellulase Synergist Gene in Sterptomyces ipomoeae

近些年来发现有些非纤维素酶组分的蛋白能协同纤维素酶系水解纤维素,大大提高纤维素酶的酶解效率,这些蛋白统称为增效蛋白。增效蛋白的发现为木质纤维素的水解提供了新的思路。本研究根据NCBI上甘薯链霉菌中潜在纤维素酶增效蛋白基因（登录号WP_009327266）设计相关引物,从甘薯链霉菌c DNA中扩增出潜在目的基因,命名为Sip1,并构建重组质粒Sip1-p ET-32a（＋）转入大肠杆菌,得到重组菌Sip1-p ET32a（＋）-E.coli BL21和Sip1-p ET32a（＋）-E.coli Rosetta（DE3）。表达的粗蛋白产物具32.3%纤维素酶增效活性。

In recent years,some non-cellulase components of the protein can be coordinated with cellulase,greatly improving the enzymatic hydrolysis efficiency of cellulose.These proteins are called synergistic proteins.The discovery of synergistic protein provides a new idea for the hydrolysis of lignocellulose.In this study,the relevant primers were designed according to the potential cellulase synergist gene（accession number WP_009327266） in Sterptomyces ipomoeae on NCBI.The potential target gene was amplified from Sterptomyces ipomoeae c DNA and named Sip1.The recombinant plasmid Sip1-p ET-32 a（＋） was constructed and transformed into E.coli.We got the recombinant bacteria Sip1-p ET32 a（＋）-E.coli BL21 and Sip1-p ET32 a（＋）-E.coli Rosetta（DE3）.The crude protein had a 32.3% cellulase synergistic activity.