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.

Decolorization of remazol brilliant blue R with laccase from Lentinus crinitus grown in agro-industrial by-products

“…It is speculated that the observed decolorization results were the outcome of distinct laccase isoenzyme expression in CH and CP media with different affinities by the substrate and/ or different catalytic properties. It is known that isoenzyme production of laccase codified by genes organized in gene families is common in fungi (Piscitelli et al 2010) and the expression of at least 10 multicopper oxidases in L. crinitus has been demonstrated (Cambri et al 2016). Moreover, the expression of laccase isoenzymes is differentially regulated and depends on the cultivation conditions as source of carbon, nitrogen and phenolic and/or aromatic compounds (Giardina et al , Pezzella et al 2013.…”

Analysis of decolorization potential of Myrothecium roridum in the light of its secretome and toxicological studies

“…lucidum allowed to reveal that proteins which can play significant bioactive roles and provide a new foundation for the further functional investigations of this fungus. However so far, only a few reports have been published on the identification of fungal extracellular proteins in the context of their later application in biodegradation processes, including the decolorization of dyes (Cambri et al 2016).

Fig.

…”

Insights into the Regulation of Algal Proteins and Bioactive Peptides Using Proteomic and Transcriptomic Approaches

“…As mentioned previously, prior to MS analysis, proteins extracted from a biological sample may be separated and digested followed by peptide sequencing using MS/MS and automated database searching. Depending on the means of protein isoelectric point and molecular weight, two-dimensional (2-D) gel electrophoresis can be used for protein separation [65]. Complementary separation steps such as liquid chromatography (LC) techniques could be used to reduce the complexity of a peptide mixture based on the physicochemical properties of peptides such as mass, charge, and hydrophobicity [66].…”