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Reference Number BB/E021271/1
Title Metabolic engineering for improved solvent production by Clostridium acetobutylicum
Status Completed
Energy Categories Renewable Energy Sources(Bio-Energy, Other bio-energy) 25%;
Not Energy Related 50%;
Renewable Energy Sources(Bio-Energy, Production of transport biofuels (incl. Production from wastes)) 25%;
Research Types Basic and strategic applied research 100%
Science and Technology Fields BIOLOGICAL AND AGRICULTURAL SCIENCES (Biological Sciences) 100%
UKERC Cross Cutting Characterisation Not Cross-cutting 100%
Principal Investigator Prof N (Nigel ) Minton
No email address given
Centre for Biomolecular Sciences
University of Nottingham
Award Type Research Grant
Funding Source BBSRC
Start Date 01 October 2007
End Date 31 March 2011
Duration 42 months
Total Grant Value £643,519
Industrial Sectors Pharmaceuticals and Biotechnology
Region East Midlands
Programme
 
Investigators Principal Investigator Prof N (Nigel ) Minton , Centre for Biomolecular Sciences, University of Nottingham (99.999%)
  Other Investigator Dr K (Klaus ) Winzer , Centre for Biomolecular Sciences, University of Nottingham (0.001%)
Web Site
Objectives Objectives not supplied
Abstract Clostridium acetobutylicum undertakes a complex biphasic fermentation: first it generates acids and then solvents, such as acetone and butanol. The project aims to generate metabolically engineered strains with altered fermentation characteristics, with the ultimate goals of (i) generating butanol hyperproducing strains and (ii) identifying the signals that govern the shift from acid to solvent formation. Single and multiple mutants of all major fermentation genes will be constructed in the sequenced strain ATTC 824. These will be characterised, initially in batch culture, with respect to their growth and fermentation characteristics. Mutants of interest will be studied in more detail: The expression of all fermentation and key glycolysis genes will be followed using quantitative RT-PCR over the entire growth period, and the concentration of relevant metabolites will be determined by LC-MS and GC-MS. The activity of key fermentation enzymes will also be monitored. Gene array analyses will be performed for key mutants at key stages of their fermentation to obtain an overall picture of the transcriptional changes occurring. In combination, this will allow us to link the individual induction time points and relative expression level of fermentation genes with the observed mutant phenotypes and to identify metabolic bottlenecks. For key mutants, the analyses described above will also be applied to chemostat-grown cultures. This will allow us to study the transcriptional, biochemical, and metabolic changes associated with the shift from acid to solvent formation under highly reproducible conditions and to dissect the specific effects of putative shift-inducing signals such as metabolite pool level, growth rate, pH, and acid concentration. Based on the above findings, a set of multiple mutants will be generated where most branches of the fermentation pathway have been inactivated (and limiting functions been over-expressed), leading to maximised butanol formation
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Added to Database 07/10/13