Abstract
The efficiency of microorganisms to degrade lignified plants is of great importance in the Earth’s carbon cycle, but also in industrial biorefinery processes, such as for biofuel production. Here, we present a large-scale proteomics approach to investigate and compare the enzymatic response of five filamentous fungi when grown on five very different substrates: grass (sugarcane bagasse), hardwood (birch), softwood (spruce), cellulose and glucose. The five fungi included the ascomycetes Aspergillus terreus, Trichoderma reesei, Myceliophthora thermophila, Neurospora crassa and the white-rot basidiomycete Phanerochaete chrysosporium, all expressing a diverse repertoire of enzymes. In this study, we present comparable quantitative protein abundance values across five species and five diverse substrates. The results allow for direct comparison of fungal adaptation to the different substrates, give indications as to the substrate specificity of individual carbohydrate-active enzymes (CAZymes), and reveal proteins of unknown function that are co-expressed with CAZymes. Based on the results, we present a quantitative comparison of 34 lytic polysaccharide monooxygenases (LPMOs), which are crucial enzymes in biomass deconstruction.
Highlights
The efficiency of microorganisms to degrade lignified plants is of great importance in the Earth’s carbon cycle, and in industrial biorefinery processes, such as for biofuel production
The fungi were grown on agar plates containing one of the five substrates and the secreted proteins were collected from the agar gel below a permeable membrane located beneath the growth site, which serves to filter away cells
The agar is homogeneous throughout the plate and the sample location is beneath the site of inoculation for all fungi, i.e. the distance of protein diffusion is equal and the protein amounts at the sample location should be comparable
Summary
The efficiency of microorganisms to degrade lignified plants is of great importance in the Earth’s carbon cycle, and in industrial biorefinery processes, such as for biofuel production. We present a quantitative comparison of 34 lytic polysaccharide monooxygenases (LPMOs), which are crucial enzymes in biomass deconstruction. Plant biomass, such as woody plants and grasses, are generally called lignocellulose because they are composed of the three main natural polymers: cellulose, hemicellulose and lignin. LPMOs have been show active on s tarch[8] In addition to these enzymes, fungi utilize oxidoreductases to modify and depolymerize lignin, including heme peroxidases, laccases, FAD-dependent oxidases and dehydrogenases[9,10,11]. The fungi secreted a large repertoire of LPMOs, the individual abundance of which varied significantly between substrates, suggesting considerable functional variation among these important enzymes
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