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Title: Regulation of Gene Expression during the Onset of Ligninolytic Oxidation by Phanerochaete chrysosporium on Spruce Wood

Source: Applied and Environmental Microbiology

Author(s)Korripally, Premsagar; Hunt, Christopher G.; Houtman, Carl J.; Jones, Don C.; Kitin, Peter J.; Cullen, Dan; Hammel, Kenneth E.; Brakhage, A. A.

Publication Year: 2015  View PDF »

Category: Journal Articles
Associated Research Project(s):   FPL-4707-2B   FPL-4709-1B   FPL-4712-2B

Abstract: Since uncertainty remains about how white rot fungi oxidize and degrade lignin in wood, it would be useful to monitor changes in fungal gene expression during the onset of ligninolysis on a natural substrate. We grew Phanerochaete chrysosporium on solid spruce wood and included oxidant-sensing beads bearing the fluorometric dye BODIPY 581/591 in the cultures. Confocal fluorescence microscopy of the beads showed that extracellular oxidation commenced 2 to 3 days after inoculation, coincident with cessation of fungal growth. Whole transcriptome shotgun sequencing (RNA-seq) analyses based on the v.2.2 P. chrysosporium genome identified 356 genes whose transcripts accumulated to relatively high levels at 96 h and were at least four times the levels found at 40 h. Transcripts encoding some lignin peroxidases, manganese peroxidases, and auxiliary enzymes thought to support their activity showed marked apparent upregulation. The data were also consistent with the production of ligninolytic extracellular reactive oxygen species by the action of manganese peroxidase-catalyzed lipid peroxidation, cellobiose dehydrogenase-catalyzed Fe3+ reduction, and oxidase-catalyzed H2O2 production, but the data do not support a role for iron-chelating glycopeptides. In addition, transcripts encoding a variety of proteins with possible roles in lignin fragment uptake and processing, including 27 likely transporters and 18 cytochrome P450s, became more abundant after the onset of extracellular oxidation. Genes encoding cellulases showed little apparent upregulation and thus may be expressed constitutively. Transcripts corresponding to 165 genes of unknown function accumulated more than 4-fold after oxidation commenced, and some of them may merit investigation as possible contributors to ligninolysis.

Keywords: Lignin biodegradation; white rot; peroxidases; reactive oxygen species; transcriptomics

Publication Review Process: Formally Refereed

Date posted: 12/15/2015

This publication is also viewable on Treesearch:  view
RITS Product ID: 77934
Current FPL Scientists associated with this product (listed alphabetically)
Hammel, Kenneth E.
Research Chemist
Houtman, Carl J.
Chemical Engineer
Hunt, Christopher G.
Research Chemist

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