title: Analysis of the Biotechnological Potential of a Lentinus crinitus Isolate in the Light of Its Secretome.
abstract: Analysis of fungal secretomes is a prospection tool for the discovery of new catalysts with biotechnological applications. Since enzyme secretion is strongly modulated by environmental factors, evaluation of growth conditions is of utmost importance to achieve optimal enzyme production. In this work, a nonsequenced wood-rotting fungus, Lentinus crinitus, was used for secretome analysis by enzymatic assays and a proteomics approach. Enzyme production was assessed after the fungus was cultured in seven different carbon sources and three nitrogen-containing compounds. The biomass yields and secreted protein arrays differed drastically among growing conditions. A mixture of secreted extracts derived from solid and liquid cultures was inspected by shotgun mass spectrometry and two-dimensional gel electrophoresis (2-DE) prior to analysis via LC-MS/MS. Proteins were identified using mass spectrometry (MS)-driven BLAST. The spectrum of secreted proteins comprised CAZymes, oxidase/reductases, proteases, and lipase/esterases. Although preseparation by 2-DE improved the number of identifications (162) compared with the shotgun approach (98 identifications), the two strategies revealed similar protein patterns. Culture media with reduced water content stimulated the expression of oxidases/reductases, while hydrolases were induced during submerged fermentation. The diversity of proteins observed within both the CAZyme and oxidoreductase groups revealed in this fungus a powerful arsenal of enzymes dedicated to the breakdown and consumption of lignocellulose.
authors: Departamento de Bioquímica, Setor de Ciências Biológicas, Centro Politécnico, Universidade Federal do Paraná , 81531-990 Curitiba-PR, Brazil.
authors: Geison
authors: G
authors: Cambri
authors: Department of Cancer Research and Molecular Medicine, Norwegian University of Science and Technology, NTNU , N-7491 Trondheim, Norway.
authors: Mirta Mittelstedt Leal
authors: MM
authors: de Sousa
authors: Department of Cancer Research and Molecular Medicine, Norwegian University of Science and Technology, NTNU , N-7491 Trondheim, Norway.|Proteomics and Metabolomics Core Facility (PROMEC), Norwegian University of Science and Technology, NTNU , N-7491 Trondheim, Norway.
authors: Davi de Miranda
authors: DM
authors: Fonseca
authors: Laboratório de Genômica Funcional, Instituto Carlos Chagas, Fundação Oswaldo Cruz , 81350-010 Curitiba-PR, Brazil.
authors: Fabricio
authors: F
authors: Marchini
authors: Departamento de Bioquímica, Setor de Ciências Biológicas, Centro Politécnico, Universidade Federal do Paraná , 81531-990 Curitiba-PR, Brazil.
authors: Joana Lea Meira
authors: JL
authors: da Silveira
authors: Departamento de Bioquímica, Setor de Ciências Biológicas, Centro Politécnico, Universidade Federal do Paraná , 81531-990 Curitiba-PR, Brazil.
authors: Jaime
authors: J
authors: Paba
mesh_terms: D018533:Biomass; D001709:Biotechnology; D004798:Enzymes / Q000032:analysis / Q000096:biosynthesis / Q000378:metabolism; D005656:Fungal Proteins / Q000302:isolation & purification* / Q000378:metabolism; D020074:Lentinula / Q000737:chemistry*; D008031:Lignin / Q000378:metabolism; D040901:Proteomics / Q000379:methods*
chemical_list: D004798:Enzymes; D005656:Fungal Proteins; C036909:lignocellulose; D008031:Lignin
keywords: 2D-PAGE; CAZyme; LC−MS; Lentinus; secretome