Development of a regulatable plasmid-based gene expression system for Clostridium thermocellum

Verfasser / Beitragende:
[Elizabeth Mearls, Daniel Olson, Christopher Herring, Lee Lynd]
Ort, Verlag, Jahr:
2015
Enthalten in:
Applied Microbiology and Biotechnology, 99/18(2015-09-01), 7589-7599
Format:
Artikel (online)
ID: 605501467
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024 7 0 |a 10.1007/s00253-015-6610-5  |2 doi 
035 |a (NATIONALLICENCE)springer-10.1007/s00253-015-6610-5 
245 0 0 |a Development of a regulatable plasmid-based gene expression system for Clostridium thermocellum  |h [Elektronische Daten]  |c [Elizabeth Mearls, Daniel Olson, Christopher Herring, Lee Lynd] 
520 3 |a Clostridium thermocellum can rapidly solubilize cellulose and produces ethanol as an end product of its metabolism. As such, it is a candidate for bioethanol production from plant matter. In this study, we developed an inducible expression system for C. thermocellum based on its native celC operon. We enhanced expression over the native operon structure by placing the repressor gene, glyR3, immediately after the celC promoter, and expressing the target gene after glyR3. Upon the addition of the inducer substrate, laminaribiose, an approximately 40-fold increase in gene expression was obtained using the test gene spo0A. Furthermore, induction of the sporulation histidine kinase, clo1313_1942, increased sporulation frequency by approximately 10,000-fold relative to an uninduced control. We have also shown that the laminaribiose (β1-3-linked carbon source) utilization pathway is not catabolite repressed by cellobiose, a β1-4-linked carbon source frequently used for C. thermocellum cultivation in laboratory conditions. Selective expression of target genes has the potential to inform metabolic engineering strategies as well as increase fundamental understanding of C. thermocellum biology. 
540 |a Springer-Verlag Berlin Heidelberg (outside the USA), 2015 
690 7 |a Laminaribiose  |2 nationallicence 
690 7 |a Inducible promoter  |2 nationallicence 
690 7 |a Biofuels  |2 nationallicence 
690 7 |a Spore formation  |2 nationallicence 
700 1 |a Mearls  |D Elizabeth  |u Thayer School of Engineering, Dartmouth College, 03755, Hanover, NH, USA  |4 aut 
700 1 |a Olson  |D Daniel  |u Thayer School of Engineering, Dartmouth College, 03755, Hanover, NH, USA  |4 aut 
700 1 |a Herring  |D Christopher  |u Thayer School of Engineering, Dartmouth College, 03755, Hanover, NH, USA  |4 aut 
700 1 |a Lynd  |D Lee  |u Thayer School of Engineering, Dartmouth College, 03755, Hanover, NH, USA  |4 aut 
773 0 |t Applied Microbiology and Biotechnology  |d Springer Berlin Heidelberg  |g 99/18(2015-09-01), 7589-7599  |x 0175-7598  |q 99:18<7589  |1 2015  |2 99  |o 253 
856 4 0 |u https://doi.org/10.1007/s00253-015-6610-5  |q text/html  |z Onlinezugriff via DOI 
898 |a BK010053  |b XK010053  |c XK010000 
900 7 |a Metadata rights reserved  |b Springer special CC-BY-NC licence  |2 nationallicence 
908 |D 1  |a research-article  |2 jats 
949 |B NATIONALLICENCE  |F NATIONALLICENCE  |b NL-springer 
950 |B NATIONALLICENCE  |P 856  |E 40  |u https://doi.org/10.1007/s00253-015-6610-5  |q text/html  |z Onlinezugriff via DOI 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Mearls  |D Elizabeth  |u Thayer School of Engineering, Dartmouth College, 03755, Hanover, NH, USA  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Olson  |D Daniel  |u Thayer School of Engineering, Dartmouth College, 03755, Hanover, NH, USA  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Herring  |D Christopher  |u Thayer School of Engineering, Dartmouth College, 03755, Hanover, NH, USA  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Lynd  |D Lee  |u Thayer School of Engineering, Dartmouth College, 03755, Hanover, NH, USA  |4 aut 
950 |B NATIONALLICENCE  |P 773  |E 0-  |t Applied Microbiology and Biotechnology  |d Springer Berlin Heidelberg  |g 99/18(2015-09-01), 7589-7599  |x 0175-7598  |q 99:18<7589  |1 2015  |2 99  |o 253