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Pull down of enzymes using xylanase activity based probes from Clostridium thermocellum

ABSTRACT: This experiment aimed to identify GH10 xylanases from Clostridium thermocellum secretome sample using activity based probes.

INSTRUMENT(S):

ORGANISM(S): Acetivibrio Thermocellus Dsm 2360

SUBMITTER: Thamy Correa

LAB HEAD: Gideon J Davies

PROVIDER: PXD065060 | Pride | 2026-06-23

REPOSITORIES: Pride

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			Action	DRS
	Clost_1_1_S1-A4_1_10766_uncalibrated.mzML	Mzml
	Clost_1_2_S1-A5_1_10767_uncalibrated.mzML	Mzml
	Clost_2_1_S1-A6_1_10768_uncalibrated.mzML	Mzml
	Clost_2_2_S1-A7_1_10769_uncalibrated.mzML	Mzml
	Clost_3_1_S1-A8_1_10770_uncalibrated.mzML	Mzml

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Project description:The members of the genus Clostridium, such as the other spore-forming anaerobic bacteria, have a complex and strictly regulated life-cycle but very little is known about genetic pathways involved at different stages. Clostridium sporogenes, Gram positive bacterium usually involved in food spoilage and frequently isolated from late blowed cheese is genetically indistinguishable from proteolytic Clostridium botulinum and is the non-neurotoxigenic counterpart often used as exemplar for the toxic subtypes. In this work we have performed a microscopy study combined with a custom array-based analysis of C. sporogenes cycle, from dormant spores to early stationary phase. We identified in spores a total of 211 transcripts validating the ipothesis that mRNAs are abundant and strikingly different from those present in growing cells. The spores transcripts included genes responsible of different life-sustaining functions, suggesting the theory of transcripts entrapment or of a basic poly-functional genes activation for future steps. In addition, after three hours from the beginning of the germination process, a 20% of the total up-regulated genes were temporally expressed in germinating spores. The vegetative condition appears to be the more active in terms of gene transcription and protein synthesis and also genes for germination and sporulation factors seem to be expressed at this point. These results suggest that spores are not silent entities and a wider knowledge about genetic pathways involved in Clostridium life cycle could be of help for a better understanding also of pathogenic clostridia types. RNA was isolated from spores, germinating spores (3h), outgrowth/transitional mid-log vegetative cells and early stationary cells of Clostridium sporogenes UC9000 previously isolated from milk. There were 3 biological replicates (independent cultures) for each condition. Probes corresponding to 3400 genomic CDSs of C. botulinum A ATCC 3502 were synthesized in three replicates randomly distributed on the chip.

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Pull down of enzymes using xylanase activity based probes from Clostridium thermocellum

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