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| Status |
Public on Feb 28, 2015 |
| Title |
Contribution of cell-bound and cell-free cellulosomal versus non-cellulosomal systems in deconstruction of cellulosic biomass by Clostridium thermocellum |
| Platform organism |
Acetivibrio thermocellus ATCC 27405 |
| Sample organism |
Acetivibrio thermocellus DSM 1313 |
| Experiment type |
Expression profiling by array
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| Summary |
The high cellulose digestion capability of the anaerobic thermophilic bacterium, Clostridium thermocellum, can be attributed to its efficient glycoside hydrolase system, consisting of both a free-enzyme system and a cellulosomal system, wherein carbohydrate active enzymes (CAZymes) are organized by primary and secondary scaffoldin proteins into large protein complexes bound to the bacterial cell wall. Using an integrated series of experiments encompassing polysaccharide depolymerization assays, direct biochemical analyses and transcriptomics, we show herein that, in addition to cell-bound cellulosomes and “free” enzymes, C. thermocellum naturally employs a system of cell-free cellulosomes that are not designed to be tethered to the cell and can diffuse away to attack polysaccharide substrates at some distance from the cell. By characterizing the cells and the secretomes of a series of mutants in which genes for either individual scaffoldins or combinations thereof are deleted, we demonstrate that the cellulosome is far more important in cellulose degradation than are the free enzymes, and that the primary scaffoldin CipA is much more important for the action of the cellulosomes than is the collective contribution of the secondary scaffoldins. Extensive transcriptomics analyses confirm the above results and extend the study to the effects of scaffoldin deletions on the organism as a whole.
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| Overall design |
To gain insights into the transcriptome profiles of C. thermocellum strains when various cellulosomal scaffoldin genes were deleted. High quality RNA was extracted from C. thermocellum grown on cellulose. Transcriptome profiles were obtained at one time point during actively growing fermentations, mid exponential phase when 50% of the substrate had been consumed.
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| Contributor(s) |
Xu Q, Podkaminer K, Yang S, Resch MG, Klingeman DM, Donohoe BS, Olson DG, Baker JO, Decker SR, Giannone RJ, Wilson CM, Hettich RL, Brown SD, Lynd LR, Bayer EA, Himmel ME, Bomble YJ |
| Citation(s) |
26989779 |
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| Submission date |
Dec 04, 2014 |
| Last update date |
Apr 01, 2016 |
| Contact name |
Charlotte Wilson |
| Organization name |
ORNL
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| Department |
Biosciences Bldg 1520
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| Lab |
329
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| Street address |
1 Bethel Valley Road
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| City |
Oak Ridge |
| State/province |
Tennessee |
| ZIP/Postal code |
37831-6342 |
| Country |
New Zealand |
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| Platforms (1) |
| GPL18170 |
NimbleGen Clostridium thermocellum ATCC 27405 HX12 array (12 x 135k)(non-condensed version) [080724_C_thermo_27405_expr] |
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| Samples (14)
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| Relations |
| BioProject |
PRJNA269315 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE63883_RAW.tar |
36.8 Mb |
(http)(custom) |
TAR (of TXT) |
| Processed data included within Sample table |
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