4.4 Article

Cloning, sequencing, and heterologous expression of a cellobiohydrolase cDNA from the basidiomycete Corticium rolfsii

Journal

BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY
Volume 67, Issue 6, Pages 1319-1326

Publisher

TAYLOR & FRANCIS LTD
DOI: 10.1271/bbb.67.1319

Keywords

cDNA; sequencing; expression; cellobiohydrolase

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From a Corticium rolfsii cDNA library, a clone homologous to other fungal cellobiohydrolase (CBH1) genes was isolated using the polymerase chain reaction. In the nucleotide sequence, one 1.6 kb long open reading frame coding for a polypeptide of 530 amino acid residues was detected which showed 64% identity with CBH1 of Phanerochaete chrysosporium. With expression of the 1.8 kb cDNA using the Aspergillus oryzae expression system, we detected microcrystalline cellulose (Avicel) hydrolyzing activity in the culture supernatant. The secreted protein, accompanied by the activity, was 89 kDa by SDS-polyacrylamide gel electrophoresis.

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