The secretome ofPleurotus sapidus

Abstract

Due to their unique capability to attack lignified biopolymers, extracellular enzymes of white-rot fungi enjoy an increasing interest in various fields of white biotechnology. The edible fungus Pleurotus sapidus was selected as a model organism for the analysis of the secretome by means of 2-DE. For enzyme production, the fungus was grown in submerged cultures either on peanut shells or on glass wool as a carrier material. Identification of the secreted enzymes was performed by tryptic digestion, ESI-MS/MS ab initio sequencing, and homology searches against public databases. The spectrum of secreted enzymes comprised various types of hydrolases and lignolytic enzymes of the manganese peroxidase/versatile peroxidase family. While peptidases were secreted mainly by the cultures grown on peanut shells, versatile peroxidase type enzymes dominated in the cultures grown on glass wool.

Quantitative Secretomic Analysis of Trichoderma reesei Strains Reveals Enzymatic Composition for Lignocellulosic Biomass Degradation

“…We have quantified 65, 37, 34, and 54 cellulolytic, hemicellulolytic, and lignin-degrading proteins and peptidases, respectively, where all were significantly higher than the corresponding 52, 16, 14, and 16 proteins that were expressed in P. chrysosporium when cultivated in cellulosic or lignin culture medium (7). The significant expression of lignocellulolytic and proteolytic enzymes were noted in T. reesei secretome when compared with those reported by fungus P. sapidus (52) and P. chrysosporium (41). Using two-dimensional gel and MALDI-TOF-MS analysis of A. oryzae (wheat bran as a major carbon source) secretome, Oda et al (53) identified 43 proteins from solid states and 37 from submerged cultures that had either lignocellulolytic or proteolytic activity.…”

Salicyl Alcohol Oxidase of the Chemical Defense Secretion of Two Chrysomelid Leaf Beetles

“…Protein spots were cut from SDS-polyacrylamide gels, washed three times with water before being digested with trypsin. The resulting peptides were subjected to electrospray ionization tandem mass spectrometry (ESI-MS/MS) as described by Zorn et al (22). For microsequencing of the N terminus, the protein was blotted onto a polyvinylidene difluoride membrane and analyzed by automated Edman degradation in an ABI Protein Sequencer 494.…”

Proteomics Methodology Applied to the Analysis of Filamentous Fungi - New Trends for an Impressive Diverse Group of Organisms

“…A reference map is a useful tool for the location of secreted proteins in addition to provide a good chance to identify those proteins. Thus, the first secretome reference maps just presented a few dozens of spots as that of P. sapidus, a white-rot fungus able to attack lignified biopolymers (Zorn et al, 2005). But the protein purification systems have evolved and several hundreds can be easily observed, as in case of T. harzianum (Suárez et al, 2005) or P. chrysogenum (Jami et al, 2010b) reference maps.…”