Record ID No. |
334 |
Author(s) |
Takakura Y., Kuwata S. , 2003 |
Affiliation |
Japan Tobacco Inc, Plant Innovation Center, Plant Breeding & Genetics Research Laboratory, 700 Higashibara Toyoda, Iwata, Shizuoka 4380802, Japan |
Title |
Purification, characterization, and molecular cloning of a pyranose oxidase from the fruit body of the basidiomycete, Tricholoma matsutake |
Source. Vol.(no):Page |
Bioscience biotechnology and biochemistry. 67(12): 2598-2607p |
Categories |
Arbuscular Mycorrhiza |
Subjects |
Biochemistry Genetics |
Sub-subjects |
Miscellaneous |
Host |
n.a. |
Organism |
Tricholoma matsutake |
Country |
Japan, Asia |
Abstracts |
A new H2O2-generating pyranose oxidase was purified as a strong antifungal protein from an arbuscular mycorrhizal fungus, Tricholoma matsutake. The protein showed a molecular mass of 250 kDa in gel filtration, and probably consisted of four identical 62 kDa subunits. The protein contained flavin moiety and it oxidized D-glucose at position C-2. H2O2 and Dglucosone produced by the pyranose oxidase reaction showed antifungal activity, suggesting these compounds were the molecular basis of the antifungal property. The V-max Km, and k(cat) for D-glucose were calculated to be 26.6 U/mg protein, 1.28 mm, and 111/s, respectively. The enzyme was optimally active at pH 7.5 to 8.0 and at 50degreesC. The preferred substrate was D-glucose, but 1,5-anhydro-D-glucitol, L-sorbose, and D-xylose were also oxidized at a moderate level. The cDNA encodes a protein consisting of 564 amino acids, showing 35.1% identity to Coriolus versicolor pyranose oxidase. The recombinant protein was used for raising the antibody |