Cloning, sequencing, and heterologous expression of a cellobiohydrolase cDNA from the basidiomycete Corticium rolfsii.
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.
Yasokawa
Daisuke
D
Section of Biotechnology, Department of Applied Technology, Hokkaido Food Processing Research Center, 589-4 Bunkyodai Midorimachi, Ebetsu, Hokkaido 069-0836, Japan. duke@foodhokkaido.gr.jp
Shimizu
Takeshi
T
Nakagawa
Ryoji
R
Ikeda
Takayuki
T
Nagashima
Koji
K
D001483:Base Sequence; D001487:Basidiomycota / Q000201:enzymology*; D043366:Cellulose 1,4-beta-Cellobiosidase / Q000096:biosynthesis / Q000235:genetics*; D003001:Cloning, Molecular; D016384:Consensus Sequence; D018076:DNA, Complementary / Q000096:biosynthesis / Q000737:chemistry; D005800:Genes, Fungal; D008969:Molecular Sequence Data; D030781:Organisms, Genetically Modified; D020075:Phanerochaete / Q000235:genetics; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D014170:Transformation, Genetic
D018076:DNA, Complementary; D043366:Cellulose 1,4-beta-Cellobiosidase