Dataset Information

Investigation of xylan degradation in Thermococcus sp. 2319x1E

ABSTRACT: Thermococcus sp. 2319x1E is able to grow on different mono- or polysaccharides as carbon source, including xylan. To investigate xylan degradation in Thermococcus sp. 2319x1E, cells were grown on 4 different carbon sources for 12 h (mid-exponential phase) and harvested by centrifugation. Using a label-free quantification approach the whole proteomes were analyzed. In another experiment a glycosidase specific activity-based probe (JJB384) was used to isolate specific Thermococcus glycosidases activated during growth on xylan or xylose.

INSTRUMENT(S): Orbitrap Fusion Lumos, LTQ Orbitrap Elite

ORGANISM(S): Unclassified Thermococcus

SUBMITTER: Farnusch Kaschani

LAB HEAD: Farnusch Kaschani

PROVIDER: PXD026056 | Pride | 2022-02-17

REPOSITORIES: Pride

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Activity-Based Protein Profiling for the Identification of Novel Carbohydrate-Active Enzymes Involved in Xylan Degradation in the Hyperthermophilic Euryarchaeon Thermococcus sp. Strain 2319x1E.

Klaus Thomas T Ninck Sabrina S Albersmeier Andreas A Busche Tobias T Wibberg Daniel D Jiang Jianbing J Elcheninov Alexander G AG Zayulina Kseniya S KS Kaschani Farnusch F Bräsen Christopher C Overkleeft Herman S HS Kalinowski Jörn J Kublanov Ilya V IV Kaiser Markus M Siebers Bettina B

Frontiers in microbiology 20220112

Activity-based protein profiling (ABPP) has so far scarcely been applied in Archaea in general and, especially, in extremophilic organisms. We herein isolated a novel Thermococcus strain designated sp. strain 2319x1E derived from the same enrichment culture as the recently reported Thermococcus sp. strain 2319x1. Both strains are able to grow with xylan as the sole carbon and energy source, and for Thermococcus sp. strain 2319x1E (optimal growth at 85°C, pH 6-7), the inducti ...[more]

PMID: 35095781

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Project description:Hemicellulose, the second most abundant plant biomass fraction after cellulose, is widely viewed as a potential feedstock for the production of liquid fuels and other value-added materials. Degradation of hemicellulose by filamentous fungi requires production of many different enzymes, which are induced by biopolymers or its derivatives and regulated mainly at the transcriptional level through transcription factors (TFs). Neurospora crassa has been shown to express and secrete plant cell wall associated enzymes. To better understand genes specifically associated with degradation of hemicellulose, we identified 353 genes by transcriptome analysis of N. crassa wild type strain grown on beechwood xylan. Exposure to xylan induces 9 of the 19 predicted hemicellulase genes. The xylanolytic phenotype of strains with deletions in genes identified from the secretome and transcriptome analysis of wild type showed that none were essential for growth on beechwood xylan. The transcription factor XlnR/Xyr1 in Aspergillus and Trichoderma species is considered to be the major transcriptional regulator of genes encoding both cellulases and hemicellulases. We identified a xlnR/xyr1 homolog in N. crassa, NCU06971, termed xlr-1 (xylanase regulator 1). Deletion of xlr-1 in N. crassa abolishes the growth on xylan and xylose, but growth on cellulose was indistinguishable from wild type. To determine regulatory mechanisms associated with hemicellulose degradation, we explored the transcriptional regulon of XLR-1 under xylose and xylanolytic versus cellulolytic conditions. XLR-1 regulated only some predicted hemicellulase genes in N. crassa and was required for a full induction of several cellulase genes. Hemicellulase gene expression was induced by a combination of release from carbon catabolite repression (CCR) and induction. However, in N. crassa, xlr-1 is subject to non-CRE-1 mediated CCR. This systematic analysis provides the similarities and differences of hemicellulose degradation and regulation mechanisms used by N. crassa in comparison to other filamentous fungi. Four-condition experiments (minimal medium, xylan medium,xylose and Avicel medium) of mutant strain(xlr-1) compared to wild type strain; Cy3 and Cy5 dye swap

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Dataset Information

Investigation of xylan degradation in Thermococcus sp. 2319x1E

Dataset's files

Publications

Activity-Based Protein Profiling for the Identification of Novel Carbohydrate-Active Enzymes Involved in Xylan Degradation in the Hyperthermophilic Euryarchaeon <i>Thermococcus</i> sp. Strain 2319x1E.

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