Session: Undergraduate Research Posters: Biochemistry:

Sustainable production of biofuels will require efficient utilization of lignocellulosic biomass. However, the current techniques are extensive and costly, so a new method must be found before biofuels can be used to their greatest potential. E. coli can be used to synthesize these biofuels; however, metabolites that accumulate during the pretreatment and fermentation processes inhibit efficiency. The goal of this project was to isolate genes from metagenomic libraries that promote survival of E. coli under acid and toxin stress. Directed evolution was performed by challenging transformed E.coli with either acetic acid or vanillin, both byproduct biofuel production. A sequence from Thermocrinis ruber, responsible for the inorganic phosphate (Pi) uptake under Pi starvation conditions, was found to promote the growth of E. coli through the vanillin trials. This gene has been isolated from its plasmid and its effects on the bacteria were monitored using an expression vector. Experimentation with acetic acid suggested that the acetate anion is the primary inhibitor. A gene was isolated that increased resistance to acetic acid and BLAST results suggest an exact match to hypothetical protein THERU_06465. The genes isolated from several experiments will be further discussed, along with their abilities to improve the production of biofuels. More trials will be conducted with the isolated genes to determine their abilities to increase toxin tolerance.

Laura Moore

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