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The Phanerochaete chrysosporium secretome: database predictions and initial mass spectrometry peptide identifications in cellulose-grown medium
KTH, School of Biotechnology (BIO).
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2005 (English)In: Journal of Biotechnology, ISSN 0168-1656, E-ISSN 1873-4863, Vol. 118, 17-34 p.Article in journal (Refereed) Published
Abstract [en]

The white rot basidiomycete, Phanerochaete chrysosporium, employs an array of extracellular enzymes to completely degrade the major polymers of wood: cellulose, hemicellulose and lignin. Towards the identification of participating enzymes, 268 likely secreted proteins were predicted using SignalP and TargetP algorithms. To assess the reliability of secretome predictions and to evaluate the usefulness of the current database, we performed shotgun LC-MS/MS on cultures grown on standard cellulose-containing medium. A total of 182 unique peptide sequences were matched to 50 specific genes, of which 24 were among the secretome subset. Underscoring the rich genetic diversity of R chrysosporium, identifications included 32 glycosyl hydrolases. Functionally interconnected enzyme groups were recognized. For example, the multiple endoglucanases and processive exocellobiohydrolases observed quite probably attack cellulose in a synergistic manner. In addition, a hemicellulolytic system included endoxylanases, alpha-galactosidase, acetyl xylan esterase, and alpha-L-arabinofuranosidase. Glucose and cellobiose metabolism likely involves cellobiose dehydrogenase, glucose oxidase, and various inverting glycoside hydrolases, all perhaps enhanced by an epimerase. To evaluate the completeness of the current database, mass spectroscopy analysis was performed on a larger and more inclusive dataset containing all possible ORFs. This allowed identification of a previously undetected hypothetical protein and a putative acid phosphatase. The expression of several genes was supported by RT-PCR amplification of their cDNAs.

Place, publisher, year, edition, pages
2005. Vol. 118, 17-34 p.
Keyword [en]
Phanerochaete chrysosporium, genome, proteome, secretome, glycosyl hydrolase, cellulase
National Category
Industrial Biotechnology
URN: urn:nbn:se:kth:diva-9010DOI: 10.1016/j.jbiotec.2005.03.010ISI: 000230328500003ScopusID: 2-s2.0-20444468865OAI: diva2:14540
QC 20101217Available from: 2006-01-16 Created: 2006-01-16 Last updated: 2010-12-17Bibliographically approved
In thesis
1. Functional genomics of wood degradation and biosynthesis
Open this publication in new window or tab >>Functional genomics of wood degradation and biosynthesis
2005 (English)Licentiate thesis, comprehensive summary (Other scientific)
Abstract [en]

Forest biotechnology is a fast emerging field of research. The application of biotechnological tools will enhance the quality of the forest products. The resultant value added and environmentally sustainable products are an absolute necessity in the future. The study of wood biosynthesis and degradation will result in enormous knowledge resources, which can be used for exploiting wood properties. This thesis addresses questions representing both wood degradation and biosynthesis.

The wood degrading fungus Phanerochaete chrysosporium is expression profiled with the microarray technology. The objective is to understand the expression pattern of the extracellular carbohydrate active enzymes (CAZymes) secreted by the organism. The data obtained increases our understanding of gene expression upon growth on cellulose.

Wood biosynthesis is studied with the model wood forming tree species, Populus. The plentiful data resources from the expression profiling during wood formation in Populus are used as the platform of this work. One of the wood specific genes, PttMAP20, previously with an unknown function is studied in this thesis. The immunolocalisation of PttMAP20 with specific antibodies is demonstrated. The putative microtubule-targeting domain of the protein is demonstrated microscopically and by using a biochemical binding assay.

Place, publisher, year, edition, pages
Stockholm: KTH, 2005. 42 p.
populus, xylogenesis, secondary cell wall, cellulose, hemicellulose, microarrays, transcript profiling
National Category
Other Biological Topics
urn:nbn:se:kth:diva-592 (URN)91-7178-250-8 (ISBN)
2006-01-30, Sal FB54, AlbaNova, Roslagstullsbacken, Stockholm, 10:00
QC 20101217Available from: 2006-01-16 Created: 2006-01-16 Last updated: 2011-11-23Bibliographically approved

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Rajangam, Alex S.Teeri, Tuula T.
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