(Pinus taeda L.) by proteomic analysis

Characterization of protein patterns from decayed wood of
loblolly pine (Pinus taeda L.) by proteomic analysis
Young-Min Young Min Kang, Kang,
Lynn Prewitt, and Susan Diehl
Department of Forest Products
Mississippi State University
IRG/WP 08-10654, 08 10654, Tuesday, May 27 th , 2008
The 39th Annual Meeting of IRG in Istanbul, Turkey

Contents
I. Introduction
II. Materials and Methods
III. Results
IV. Conclusion

I. Introduction
Biodeterioration is the undesirable change of
the properties of a material by an organism
Wood decay: the loss of billions of US dollars annually
The main fungi of wood decay: Basidiomycete
Traditional methods for protecting wood: chemical preservatives

I. Introduction (continued)
Identification of basidiomycetes and
proteins (enzymes) on wood
What kind of microbes present on wood?
Not all microbes are actively decaying the
wood
When organisms are metabolically active,
such as during wood decay, they produce
proteins

I. Introduction (continued)
� Expressional proteomics can provide a snap shot of all
proteins expressed by an organism at the time of extraction.
� During the process of decay, fungi express different
proteins involved in both the decay of wood and metabolism
of the fungus.
� OBJECTIVE of this study is to evaluate protein expression
in decay fungi growing on loblolly pine wood

II. Materials and Methods
Fresh Wood Decayed Wood Inoculated Wood
GT
Gloeophyllum trabeum

II. Materials and Methods (continued)
• IEF and 2DE (Two-dimensional gel electrophoresis)

II. Materials and Methods (continued)
Excise Protein Spots and ID by Mass Spec.
Direct Spotting onto
MALDI target
GPS Explorer
Software for Data
Analysis
4800 MALDI TOF/TOF TM
Analyzer
MS/MS Analysis
MS Analysis

III. Results
FW
Fresh Wood = No Proteins Detected

III. Results (continued)
DW
Decayed wood = 1 Protein identified as Actin

III. Results (continued)
IW
Inoculated wood with G. trabeum

III. Results (continued)
Inoculated wood (IW):
� 76 Total proteins detected
� 5 Unidentified (incomplete peptides)
� 54 Hypothetical proteins (unknown proteins)
� 17 Identified
Of those identified: 3 decay enzymes
alcohol oxidase
lipoxygenase
catalase

III. Results (continued)
GT
Pure culture of G. trabeum

III. Results (continued)
From Pure Culture of GT :
� 110 Total proteins detected
� 8 Unidentified (incomplete peptides)
� 57 Hypothetical proteins (unknown proteins)
� 45 Identified
Including actin, heat shock protein, cell division
related enzymes and general metabolic enzymes

IV. Conclusion
This is the first reported proteomic analysis of
decayed wood and provides a useful tool for
characterization of proteins involved in wood
biodeterioration.
GT on wood needs wood decay enzymes to break
down the wood for food while GT in culture does not
need these enzymes to obtain the food.
This type of comparison will help us understand the
complex mechanism of decay, in particular it will
allow us to discover new proteins not previously
known to play a role in decay.

Conclusion (continued)
This study also highlights one of the current limitations
to fungal proteomic studies, the lack of identified fungal
proteins in the available databases.
This problem parallels that which occurred in years
past with the lack of sequence information on GenBank.
We plan to create our own wood decay protein
database.

Conclusion (continued)

ACKNOWLEDGMENTS
Dr. Darrel Nicholas and Dr. Tor Schultz
at Mississippi State University
Dr. Tibor Pechan
(Life Sciences and Biotechnology Institute)
Grant Kirker, Robert Bucci, Lee Mangum, and Min Lee

Questions ?
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