Fungal degradation of wood: Initial proteomic analysis of extracellular proteins of Phanerochaete chrysosporium grown on oak substrate

Ahmed Abbas, Hasan Koc, Feng Liu, Ming Tien

Research output: Contribution to journalArticle

79 Citations (Scopus)

Abstract

Two-dimensional (2-D) gel electrophoresis was used to separate the extracellular proteins produced by the white-rot fungus Phanerochaete chrysosporium. Solid-substrate cultures grown on red oak wood chips yielded extracellular protein preparations which were not suitable for 2-D gel analysis. However, pre-washing the wood chips with water helped decrease the amount of brown material which caused smearing on the acidic side of the isoelectric focusing gel. The 2-D gels from these wood-grown cultures revealed more than 45 protein spots. These spots were subjected to in-gel digestion with trypsin followed by either peptide fingerprint analysis by matrix assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF/MS) or by liquid chromatography (LC)/MS/MS sequencing. Data from both methods were analyzed by Protein Prospector and the local P. chrysosporium annotated database. MALDI-TOF/MS only identified two proteins out of 25 analyzed. This was most likely due to problems associated with glycosylation. Protein sequencing by LC/MS/MS of the same 25 proteins resulted in identification of 16 proteins. Most of the proteins identified act on either cellulose or hemicellulose or their hydrolysis products. Thus far no lignin peroxidase, Mn peroxidase or laccases have been detected.

Original languageEnglish (US)
Pages (from-to)49-56
Number of pages8
JournalCurrent Genetics
Volume47
Issue number1
DOIs
StatePublished - Jan 1 2005

Fingerprint

Phanerochaete
Proteomics
Proteins
Gels
manganese peroxidase
Liquid Chromatography
Mass Spectrometry
Lasers
Laccase
Peptide Mapping
Protein Sequence Analysis
Electrophoresis, Gel, Two-Dimensional
Isoelectric Focusing
Glycosylation
Cellulose
Trypsin
Digestion
Hydrolysis
Fungi
Databases

All Science Journal Classification (ASJC) codes

  • Genetics

Cite this

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abstract = "Two-dimensional (2-D) gel electrophoresis was used to separate the extracellular proteins produced by the white-rot fungus Phanerochaete chrysosporium. Solid-substrate cultures grown on red oak wood chips yielded extracellular protein preparations which were not suitable for 2-D gel analysis. However, pre-washing the wood chips with water helped decrease the amount of brown material which caused smearing on the acidic side of the isoelectric focusing gel. The 2-D gels from these wood-grown cultures revealed more than 45 protein spots. These spots were subjected to in-gel digestion with trypsin followed by either peptide fingerprint analysis by matrix assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF/MS) or by liquid chromatography (LC)/MS/MS sequencing. Data from both methods were analyzed by Protein Prospector and the local P. chrysosporium annotated database. MALDI-TOF/MS only identified two proteins out of 25 analyzed. This was most likely due to problems associated with glycosylation. Protein sequencing by LC/MS/MS of the same 25 proteins resulted in identification of 16 proteins. Most of the proteins identified act on either cellulose or hemicellulose or their hydrolysis products. Thus far no lignin peroxidase, Mn peroxidase or laccases have been detected.",
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Fungal degradation of wood : Initial proteomic analysis of extracellular proteins of Phanerochaete chrysosporium grown on oak substrate. / Abbas, Ahmed; Koc, Hasan; Liu, Feng; Tien, Ming.

In: Current Genetics, Vol. 47, No. 1, 01.01.2005, p. 49-56.

Research output: Contribution to journalArticle

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