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

Korripally, Premsagar; Hunt, Christopher G.; Houtman, Carl J.; Jones, Don C.; Kitin, Peter J.; Cullen, Dan; Hammel, Kenneth E.

doi:10.1128/AEM.02064-15

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

Journal Article · Fri Sep 04 00:00:00 EDT 2015 · Applied and Environmental Microbiology

DOI:https://doi.org/10.1128/AEM.02064-15· OSTI ID:1470155

Korripally, Premsagar ^[1]; Hunt, Christopher G. ^[2]; Houtman, Carl J. ^[2]; Jones, Don C. ^[2]; Kitin, Peter J. ^[2]; Cullen, Dan ^[3]; Hammel, Kenneth E. ^[3]

Mahatma Gandhi Univ., Nalgonda (India). Dept. of Biotechnology
Forest Products Lab., Madison, WI (United States)
Forest Products Lab., Madison, WI (United States); Univ. of Wisconsin, Madison, WI (United States). Dept. of Bacteriology

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. In this paper, 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. Finally, 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.

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Research Organization:: Univ. of Wisconsin, Madison, WI (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Biological and Environmental Research (BER)

Grant/Contract Number:: SC0006929

OSTI ID:: 1470155

Journal Information:: Applied and Environmental Microbiology, Vol. 81, Issue 22; ISSN 0099-2240

Publisher:: American Society for MicrobiologyCopyright Statement

Country of Publication:: United States

Language:: English

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Cited by: 35 works

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