Novel cellulase profile of Trichoderma reesei strains constructed by cbh1 gene replacement with eg3 gene expression cassette.
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.
Wang
Tian-Hong
TH
The State Key Laboratory of Microbial Technology, Shandong University, Jinan 250100, China. wangtianhong@sdu.edu.cn.
Liu
Ti
T
Wu
Zhi-Hong
ZH
Liu
Shi-Li
SL
Lu
Yi
Y
Qu
Yin-Bo
YB
D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D002480:Cellulase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D043366:Cellulose 1,4-beta-Cellobiosidase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D004789:Enzyme Activation; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology; D015966:Gene Expression Regulation, Fungal / Q000502:physiology; D020868:Gene Silencing / Q000502:physiology; D011401:Promoter Regions, Genetic; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000378:metabolism; D013045:Species Specificity; D014242:Trichoderma / Q000145:classification / Q000201:enzymology* / Q000235:genetics*
D001426:Bacterial Proteins; D011994:Recombinant Proteins; D002480:Cellulase; C061269:cel-3 protein, Fibrobacter succinogenes; D043366:Cellulose 1,4-beta-Cellobiosidase