Novel Cellulase Profile of <italic>Trichoderma reesei</italic> Strains Constructed by <italic>cbh1</italic> Gene Replacement with <italic>eg3</italic> Gene Expression Cassette

作者：Tianhong Wang, Ti Liu, Zhihong Wu, Shili Liu, Yi Lu, Yinbo Qu · 发表于：Acta Biochimica et Biophysica Sinica · 年份：2004 · DOI：10.1093/abbs/36.10.667 · 被引用次数：18 · 研究领域：Biofuel production and bioconversion、Enzyme Production and Characterization、Microbial Metabolic Engineering and Bioproduction

To construct strains of the filamentous fungus Trichoderma reesei with low cellobiohydrolases while high endoglucanase activity, the P(cbh1)-eg3-T(cbh1) cassette was constructed and the coding sequence of the cellobiohydrolase I (CBHI) gene was replaced with the coding sequence of the eg3 gene by homologous recombination. Disruption of the cbh1 gene was confirmed by PCR, Southern dot blot and Western hybridization analysis in two transformants denoted as L13 and L29. The filter paper-hydrolyzing activity of strain L29 was 60% of the parent strain Rut C30, and the CMCase activity was increased by 33%. This relatively modest increase suggested that the eg3 cDNA under the control of the cbh1 promoter was not efficiently transcribed as the wild type cbhl gene. However our results confirmed that homologous recombination could be used to construct strains of the filamentous fungus Trichoderma reesei with novel cellulase profile. Such strains are of interest from the basic science perspective and also have potential in ustrial applications.