Abstract
A pyranose 2-oxidase gene from the brown-rot basidiomycete Gloeophyllum trabeum was isolated using homology-based degenerate PCR. The gene structure was determined and compared to that of several pyranose 2-oxidases cloned from white-rot fungi. The G. trabeum pyranose 2-oxidase gene consists of 16 coding exons with canonical promoter CAAT and TATA elements in the 5’UTR. The corresponding G. trabeum cDNA was cloned and contains an ORF of 1,962 base pairs encoding a 653 amino acid polypeptide with a predicted molecular weight of 72 kDa. A Hisx6 tagged recombinant G. trabeum pyranose 2-oxidase was generated and expressed heterologously in Escherichia coli yielding 15 U enzyme activity per ml of induced culture. Structural alignment and phylogenetic analysis were performed and are discussed.
Keywords
Gene cloning,
Gloeophyllum trabeum,
glucose oxidase,
pyranose 2-oxidase,
wood-decaying fungi,
fungi,
biotechnology,
industrial applications,
Basidiomycetes,
cloning,
microbial metabolism,
gene expression,
genetic engineering,
genetics,
molecular genetics,
enzymes,
recombinant DNA,
amino acids,
polypeptides,
Escherichia coli,
phylogeny,
brown rot,
oxidation,
decay fungi,
CAAT,
TATA,
genetic analysis
Citation
Dietrich, Diane; Crooks, Casey. 2009. Gene cloning and heterologous expression of pyranose 2-oxidase from the brown-rot fungus, Gloeophyllum trabeum. Biotechnology letters. Vol. 31, no. 8 (Aug. 2009): pages 1223-1228.
