The genes encoding glycoside hydrolase family 6 and 7 cellulases from the brown-rot fungus Coniophora puteana

Abstract

Four genes encoding glycoside hydrolase (GH) family 6 and 7 cellulases (cel6A, cel6B, cel7A, and cel7B) were obtained from the brown-rot fungus Coniophora puteana by genomic polymerase chain reaction (PCR) using consensus degenerate hybrid oligonucleotide primers (CODEHOPs) designated from the amino acid sequence of cellobiohydrolases (CBHs) from white-rot fungi. The nucleotide sequences of four genes showed high homology with basidiomycetes CBHs, suggesting the fi rst cloning of the genes encoding Cel6 and Cel7 from brown-rot fungi. PCR using CODEHOP pairs at the catalytic domain successfully amplifi ed both cel6A and cel6B, whereas only cel6A fragment was obtained using the primers including the carbohydrate-binding modules (CBMs), suggesting lack of CBM in Cel6B. Moreover, both cel7A and cel7B were amplified by the PCR using CODEHOP pairs at the catalytic domain, but not by those including CBM, suggesting the absence of Cel7 with CBM in the fungus. From these results, three of four cellulases from C. puteana may not carry CBM, which has an important role for the degradation of crystalline cellulose.