Abstract
To identify the extracellular enzymes produced by Enoki-take mushrooms (Flammulina velutipes), we performed secretome analysis using transcriptomic sequence database of the fungus. F.velutipes was cultivated using various types of biomass and related polysaccharides as carbon sources and total RNA was extracted from the mycelia obtained from each culture. The normalized cDNA library constructed from the total RNA was sequenced by 2nd generation DNA sequencer and subsequently a transcriptomic sequence database of F.velutipes was constructed. The extracellular proteins produced by the fungus in the cellulose-degrading culture were separated by 2-dimensional gel electrophoresis and the obtained spots were identified using the transcriptomic sequence database. A total of 41 protein spots were assigned to corresponding contig sequences, and the various enzymes related to carbohydrate degradation were identified, suggesting the validity of the database for the secretome analysis without total genomic sequence or genome-wide annotation.
