Differential Expression in Phanerochaete chrysosporium of Membrane-Associated Proteins Relevant to Lignin Degradation
Fungal lignin-degrading systems must include membrane-associated proteins that participate in diverse processes such as uptake and oxidation of lignin fragments, secretion of ligninolytic secondary metabolites, and defense of the mycelium against ligninolytic oxidants. Despite their importance, little is known about the nature or regulation of these membrane-associated components. We grew the white rot basidiomycete Phanerochaete chrysosporium on cellulose or glucose as the carbon source and monitored the mineralization of a 14C-labeled synthetic lignin by these cultures to assess their ligninolytic competence. The results showed that the cellulose-grown cultures were ligninolytic, whereas the glucose-grown ones were not. We isolated microsomal membrane fractions from both types of culture and analyzed tryptic digests of them by shotgun liquid chromatography/tandem mass spectrometry. Comparison of the results against the predicted P. chrysosporium proteome showed that a catalase (Joint Genome Institute P. chrysosporium protein I.D. 124398), an alcohol oxidase (126879), two transporters (137220 and 132234), and two cytochrome P450s (5011 and 8912) were up-regulated under ligninolytic conditions. Real time reverse transcription polymerase chain reaction assays showed that RNA transcripts encoding all of these proteins were also up-regulated in ligninolytic cultures. Catalase 124398, alcohol oxidase 126879, and transporter 137220 were found in a proteomic analysis of partially purified plasma membranes from ligninolytic P. chrysosporium, and are therefore most likely associated with the outer envelope of the fungus.
- Research Organization:
- Pacific Northwest National Lab. (PNNL), Richland, WA (United States). Environmental Molecular Sciences Lab. (EMSL)
- Sponsoring Organization:
- USDOE
- DOE Contract Number:
- AC05-76RL01830
- OSTI ID:
- 974994
- Report Number(s):
- PNNL-SA-63951; AEMIDF; 28293; BM0102070; TRN: US201007%%932
- Journal Information:
- Applied and Environmental Microbiology, 74(23):7252-7257, Vol. 74, Issue 23; ISSN 0099-2240
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
CARBON SOURCES
CATALASE
CELLULOSE
CYTOCHROMES
GLUCOSE
LIGNIN
MASS SPECTROSCOPY
MEMBRANES
METABOLITES
MINERALIZATION
MYCELIUM
OXIDASES
OXIDATION
OXIDIZERS
PHANEROCHAETE
POLYMERASE CHAIN REACTION
PROTEINS
RNA
SECRETION
TRANSCRIPTION
Phanerochaete chrysosporium
lignin
Environmental Molecular Sciences Laboratory