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Method for characterizing extracellular proteins from the cell wall proteome of the copper tolerant fungus Phialophora malorum

Daniel, Geoffrey and Volc, Jindrich and Halada, Petr and Karunasekera, Hasanthi and Kim, Jong Sik (2019). Method for characterizing extracellular proteins from the cell wall proteome of the copper tolerant fungus Phialophora malorum. International Biodeterioration and Biodegradation. 144 , 104769
[Research article]

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Abstract

Phialophora malorum is a well-known copper tolerant fungus causing soft rot decay of copper-treated wood in-service in ground contact situations worldwide. The aim of the present work was to develop a method that allowed studies of the mechanism(s) behind extracellular copper tolerance by examining the profile of proteins expressed by P. malorum in the cell wall/slime (subproteome) environment in mycelia grown in Cu-supplemented media. The study involves development of a novel non-destructive approach using mild extraction, concentration, separation (SDS-PAGE electrophoresis) and characterization (MALDI-TOF MS/MS) of the proteins present. Studies revealed differential protein profiles and expression in Cu-supplemented (0.1% CuSO4) vs control shake cultures after 10 days growth. Twelve proteins including several hydrolytic (i.e. glucosidases, pectinase, polygalacturonase) enzymes were either unique or over-expressed in Cu-cultures compared to controls. The cell wall location of the proteins was consistent with a role in copper tolerance and detoxification by binding Cu-ions extracellularly. Several of the proteins (e.g. hydrolases, pectinase) are also involved in wood cell wall biomineralization. It is proposed that the concentration of metabolites (organic acids) slowly released in close vicinity of hyphal surfaces by weakly bound enzymes over-expressed through Cu-stress may function to reduce levels of free non-chelated Cu (II) ions entering the cell cytoplasm and therefore provide a first level of defense against copper toxicity.

Authors/Creators:Daniel, Geoffrey and Volc, Jindrich and Halada, Petr and Karunasekera, Hasanthi and Kim, Jong Sik
Title:Method for characterizing extracellular proteins from the cell wall proteome of the copper tolerant fungus Phialophora malorum
Series Name/Journal:International Biodeterioration and Biodegradation
Year of publishing :2019
Volume:144
Article number:104769
Number of Pages:7
ISSN:0964-8305
Language:English
Publication Type:Research article
Article category:Scientific peer reviewed
Version:Published version
Full Text Status:Public
Subjects:(A) Swedish standard research categories 2011 > 1 Natural sciences > 106 Biological Sciences (Medical to be 3 and Agricultural to be 4) > Microbiology (Microbiology in the medical area to be 30109)
(A) Swedish standard research categories 2011 > 4 Agricultural Sciences > 401 Agricultural, Forestry and Fisheries > Wood Science
Keywords:Copper tolerance, Differential protein profiles, MALDI-TOF MS/MS, Peptide sequencing, Phialophora malorum, Soft rot decay, Cell wall proteome
URN:NBN:urn:nbn:se:slu:epsilon-p-101433
Permanent URL:
http://urn.kb.se/resolve?urn=urn:nbn:se:slu:epsilon-p-101433
Additional ID:
Type of IDID
DOI10.1016/j.ibiod.2019.104769
Web of Science (WoS)000491619000023
ID Code:24618
Faculty:S - Faculty of Forest Sciences
Department:(S) > Department of Forest Biomaterials and Technology
Deposited By: SLUpub Connector
Deposited On:21 Jun 2021 14:59
Metadata Last Modified:21 Jun 2021 15:01

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