Analysis of the Biotechnological Potential of a Lentinus crinitus Isolate in the Light of Its Secretome
| Autor: | Mirta M. L. Sousa, Fabricio K Marchini, Geison Cambri, Jaime Paba, Joana Léa Meira Silveira, Davi de Miranda Fonseca |
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| Rok vydání: | 2016 |
| Předmět: |
Proteomics
0106 biological sciences 0301 basic medicine Proteases Lentinula Lignin 01 natural sciences Biochemistry Fungal Proteins 03 medical and health sciences Liquid chromatography–mass spectrometry 010608 biotechnology Biomass Lipase chemistry.chemical_classification Gel electrophoresis biology General Chemistry biology.organism_classification Enzymes 030104 developmental biology Enzyme Secretory protein chemistry biology.protein Lentinus Biotechnology |
| Zdroj: | Journal of Proteome Research. 15:4557-4568 |
| ISSN: | 1535-3907 1535-3893 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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