cellulase and methylglucoside

We have investigated the effect of disruption of the bgl1-(beta-glucosidase l-encoding) gene of Trichoderma reesei on the formation of other beta-glucosidase activities and on the induction of cellulases. To this end the bgl1 locus was disrupted by insertion of the Aspergillus nidulans amdS (acetamidase-encoding) gene. The bgl1-disrupted strain did not produce the 75 kDa extracellular beta-glucosidase on cellulose or lactose, but still formed beta-glucosidase activity on glucose, cellobiose, xylan or beta-1,3-glucan, suggesting that the enzyme(s) exhibiting this beta-glucosidase activity is (are) not encoded by bgl1. The cellulase-inducer sophorose induced the bgl1-encoded beta-glucosidase, whereas the remaining beta-glucosidase activity was induced by methyl-beta-D-glucoside. The bgl1-gene product was mainly secreted into the medium, whereas the other beta-glucosidase activity was mainly associated with the cells. A bgl1-multicopy strain formed higher amounts of cellulases than the parent strain. Nonsaturating concentrations of sophorose efficiently induced cellobiohydrolase l formation in the bgl1-multicopy strain, but less efficiently in the bgl1-disrupted strain. The multicopy strain and the parent strain were comparably efficient at saturating sophorose concentrations. The beta-glucosidase inhibitor nojirimycin strongly inhibited induction in all strains. These data suggest that the bgl1-encoded beta-glucosidase is not identical to the plasma-membrane-bound, constitutive, methyl-beta-glucoside inducible beta-glucosidase, but represents an extracellular cellulose-induced enzyme. Both enzymes contribute to rapid induction of cellulases by modifying the inducer sophorose.

Topics: Base Sequence; beta-Glucosidase; Cellulase; Cellulose; Cloning, Molecular; DNA Primers; DNA, Fungal; Enzyme Induction; Gene Amplification; Gene Expression; Genes, Fungal; Glucans; Methylglucosides; Molecular Sequence Data; Promoter Regions, Genetic; Trichoderma

Transcription of the Trichoderma longibrachiatum egl1 gene is induced in the presence of lactose and beta-methylglucoside and repressed by glucose. A DNA fragment containing 722 bp upstream of the ATG codon has been sequenced. The gene has two major transcription start points (20 and 24 nucleotides upstream from the ATG codon) and several transcription termination points (located in a region around 130 nt downstream of the stop codon). Two 6-mer sequences (5'-CTGGAG-3') separated by 16 bp are present in the egl1 gene promoter. These sequences match the Aspergillus nidulans consensus CreA binding site and might be implicated in carbon catabolite repression of egl1 transcription.

Topics: Base Sequence; Binding Sites; Cellulase; Enzyme Repression; Fungal Proteins; Gene Expression Regulation, Fungal; Genes, Fungal; Lactose; Methylglucosides; Molecular Sequence Data; Promoter Regions, Genetic; Repressor Proteins; Terminator Regions, Genetic; Transcription, Genetic; Trichoderma