Biofuels Production projects

Engineering Phage Resistance in Solventogenic Clostridia Using CRISPR/Cas

Bacteriophages, viruses that infect bacteria, have the potential to interfere with any industrial process that relies on bacterial fermentation including the acetone-butanol-ethanol (ABE) fermentation with solventogenic clostridia. During the history of the ABE fermentation, there have been several documented bacteriophage (phage) infections resulting in dramatically reduced yields. It is prudent to expect that phage infections will continue with the redevelopment of the ABE fermentation industry. The traditional method to deal with phage infections is to isolate bacteriophage insensitive mutants (BIMs) using a process termed "immunization." However, this strategy is not ideal because it 1) only treats the current phage infection; 2) takes a minimum of 2 weeks; and 3) often results in BIMs with reduced fermentation characteristics compared to the parental strain. To insure the profitability of the ABE industry, it is necessary to develop an economical solution to phage infections.

project Highlights

2014 Highlights

Bacteriophages -- viruses that infect bacteria -- can potentially interfere with industrial processes such as the acetone-butanol-ethanol (ABE) fermentation with solventogenic clostridia. The goals of this project are to (1) identify and characterize solventogenic clostridial phages and (2) evaluate non-CRISPR phage resistance strategies. We have made progress on both goals. We have characterized two phages for C. saccharoperbutylacetonicum N1-4 and will continue to search for more phages to this strain and others. We have also identified several bacteriophage-insensitive mutants (BIMs) and will conduct further analyses on these in the future.




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