冬小麦膨胀素基因EXPB7工程菌构建及纤维素水解作用分析

Construction of Engineering Strain of Winter Wheat Swollenin Gene EXPB7 and Analysis of Cellulose Hydrolysis

为了寻找促进纤维素降解的有效方法来提高纤维素酶的降解效率。本研究以东农冬麦1号膨胀素基因EXPB7为供体,构建黑曲霉表达载体,以黑曲霉CICC2462为受体,采用农杆菌介导法遗传转化黑曲霉,构建膨胀素基因黑曲霉工程菌pSZHGS-EXPB7,并对该工程菌发酵液的纤维素水解促进作用进行分析。结果表明,黑曲霉重组膨胀素蛋白在对滤纸处理2 d后崩解效果明显,与出发菌株的分泌蛋白相比,具有一定的促进作用;重组菌株发酵液上清处理滤纸产生的还原糖含量显著高于出发菌株(P<0.05),与仅用酶液处理滤纸相比,能够将纤维素酶的水解效率提高32.9%。

In order to find an effective method for promoting cellulose degradation to improve the degradation efficiency of cellulase. In this study,the Aspergillus oryzae EXPB7 was used as a donor to construct the Aspergillus niger expression vector,and the Aspergillus niger CICC2462 was used as a receptor to genetically trans form Aspergillus niger by Agrobacterium-mediated transformation to construct the swollenin gene Aspergillus niger. pSZHGS-EXPB7,and the cellulose hydrolysis promoting effect of the fermentation broth of the engineering bacteria was analyzed. The results showed that the Aspergillus niger recombinant swollenin protein had obvious disintegration effect after 2 days of treatment on the filter paper,and it had a certain promoting effect compared with the secreted protein of the original strain. The glucose content of the supernatant of the recombinant strain fermentation supernatant was significantly higher than that of the original strain(P<0.05),and the hydrolysis efficiency of the cellulase was increased by 32.9% compared with the filter paper treated with the enzyme solution alone.