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Isolation and Preparation of Extracellular Proteins from Lignocellulose Degrading Fungi for Comparative Proteomic Studies Using Mass Spectrometry

  • Robert J. Gruninger
  • Adrian Tsang
  • Tim A. McAllister
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1588)

Abstract

Fungi utilize a unique mechanism of nutrient acquisition involving extracellular digestion. To understand the biology of these microbes, it is important to identify and characterize the function of proteins that are secreted and involved in this process. Mass spectrometry-based proteomics is a powerful tool to study complex mixtures of proteins and understand how the proteins produced by an organism change in response to different conditions. Many fungi are efficient decomposers of plant cell wall, and anaerobic fungi are well recognized for their ability to digest lignocellulose. Here, we outline a protocol for the enrichment and isolation of proteins secreted by anaerobic fungi after growth on simple (glucose) and complex (straw and alfalfa hay) carbon sources. We provide detailed instruction on generating protein fragments and preparing these for proteomic analysis using reversed phase chromatography and mass spectrometry.

Key words

Exo-proteome Fungi CAZy Proteomics Lignocellulose Mass spectrometry 

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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  • Robert J. Gruninger
    • 1
  • Adrian Tsang
    • 2
  • Tim A. McAllister
    • 1
  1. 1.Lethbridge Research and Development CentreAgriculture and Agri-Food CanadaLethbridgeCanada
  2. 2.Centre for Structural and Functional GenomicsConcordia UniversityQCCanada

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