| Popis: |
Industrial and agricultural wastes such as sugarcane bagasse (BCA) and crop-shells are abundant in Brazil and, in this view, represent important substrates for biotechnology purposes. Previously, we have demonstrated that the thermophilic fungus Humicola grisea var. thermoidea, when cultivated in lignocellulosic materials such as BCA and ball-milled straw (BMS), secretes several hydrolytic enzymes resulting in a complex proteic profile. Among others, in this enzymatic pool we detected a highly expressed protein with a molecular mass of about 50 kDa. In the present work, this major component was identified by distinct experimental approaches such as immunoblot, isoelectric-focusing and automatic sequencing of tryptic peptides. Amino acids homology analyses identified this protein (CBH1.2) as corresponding to the cellobiohydrolase 1.2 gene (cbh1.2) product. The CBHI.2 protein presented a molecular mass of 49.6 kDa and a pI of 5.27. The full-length cbh1.2 cDNA was obtained through reverse transcriptase–PCR (RT–PCR) in order to be expressed in different heterologous hosts. Taken together, our results point to the relevance of CBHI.2 in the H. grisea hydrolytic system and to the potential industrial application of its gene product and the highly induced culture supernatant. |
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