Abstract
White-rot fungi secret a large number of hydrolytic and oxidative enzymes for degradation of lignocellulosic material. The sequencing of the genome of the white-rot fungus Phanerochaete chrysosporium has facilitated the characterization of its complete extracellular proteome. P. chrysosporium was grown on liquid medium, containing glucose, cellulose or wood chips as the carbon source, and also in solid substrate fermentation bags. For liquid-grown cultures, the extracellular protein fraction was separated by 2D gel electrophoresis. Protein spots were analysed by in-gel digestion and liquid chromatography (LC)/MS/MS. A total of 18 additional protein spots from the 2D gels yielded hits from blast searches. From solid substrate cultures in which the fungus was grown in bags, the proteins were resolved by SDS-PAGE, subjected to in-gel digestion and then identified by LC/MS/MS. An additional 16 proteins yielded hits on blast searches. Enzymes involved in cellulose, hemicellulose, lignin and protein degradation were identified. Expression patterns were very similar between cellulose-grown cultures and wood-grown cultures. In addition to enzymes which act on lignocellulosic material, proteases were also found, indicating the need of fungi to scavenge for nitrogen in wood.
Highlights
Woody biomass is a complex mixture of cellulose, hemicellulose and lignin
Using 2D gels with in-gel digestion and liquid chromatography (LC)/MS/MS, we previously identified 16 proteins from the extracellular proteome of P. chrysosporium
P. chrysosporium was again grown on submerged wood medium with the objective of determining the extracellular proteome (Fig. 1)
Summary
Woody biomass is a complex mixture of cellulose, hemicellulose and lignin. Enzymes involved in degradation of these two polymers must hydrolyse the glycosidic bonds that join the monomeric subunits, and the sugars which modify the hemicellulose (Kirk & Cullen, 1998). These side groups render hemicellulose non-crystalline because they prevent efficient packing of hemicellulose. Lignin is the most complex of the three polymers and can constitute over 25 % by weight of woody biomass (Adler, 1977). The complexity of lignin arises from the 12 different types of linkages that link the monomers (Adler, 1977)
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