Abstract
Proteins secreted by the white-rot, softwood-degrading fungus Phanerochaete carnosa during growth on cellulose and spruce were analyzed using tandem mass spectrometry and de novo sequencing. Homology-driven proteomics was applied to compare P. carnosa peptide sequences to proteins in Phanerochaete chrysosporium using MS BLAST and non-gapped alignment. In this way, 665 and 365 peptides from cellulose and spruce cultivations, respectively, were annotated. Predicted activities included endoglucanases from glycoside hydrolase (GH) families 5, 16, and 61, cellobiohydrolases from GH6 and GH7, GH3 β-glucosidases, xylanases from GH10 and GH11, GH2 β-mannosidases, and debranching hemicellulases from GH43 and CE15. Peptides corresponding to glyoxal oxidases, peroxidases, and glycopeptides that could participate in lignin degradation were also detected. Overall, predicted activities detected in extracellular filtrates of cellulose and spruce cultures were similar, suggesting that the adaptation of P. carnosa to growth on lignocellulose might result from fine tuning the expression of similar enzyme families.
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Acknowledgments
This work was supported by grants from the Natural Sciences and Engineering Research Council to E.R.M. We would like to acknowledge Dr. E. Yang (Proteomics Core Facility, Sunnybrook Research Institute) for sharing his expertise in proteomic analyses and critical review of the manuscript. We also thank Mr. T. Rimstad for kindly sharing his expertise in Python programming and Dr. I. Adzhubey (Division of Genetics, Harvard Medical School) for his assistance with MS BLAST.
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Supplementary Table 1
Non-gapped alignment of peptides from extracellular filtrates of P. carnosa grown on cellulose and spruce to probable lignocellulose-degrading enzymes from P. chrysosporium (DOC 105 kb)
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Mahajan, S., Master, E.R. Proteomic characterization of lignocellulose-degrading enzymes secreted by Phanerochaete carnosa grown on spruce and microcrystalline cellulose. Appl Microbiol Biotechnol 86, 1903–1914 (2010). https://doi.org/10.1007/s00253-010-2516-4
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DOI: https://doi.org/10.1007/s00253-010-2516-4