Location: Renewable Product Technology Research
Project Number: 5010-41000-135-04-R
Project Type: Reimbursable Cooperative Agreement
Start Date: Dec 15, 2009
End Date: Jun 14, 2014
1. Characterize the spectrum of activity of phage endolysins against our collection of bacterial contaminants from fuel ethanol plants. 2. Mutatgenesis/directed evolution to broaden the spectrum of activity of endolysins. 3. Clone endolysin genes for functional expression in a cellulosic microbial catalyst. 4. Demonstrate efficacy of endolysins in ethanol fermentations.
Fuel ethanol is not produced under aseptic conditions, and acute bacterial infections occur unpredictably leading to "stuck" fermentations. We propose developing a cellulosic yeast ethanologen that secretes a bactericidal lytic enzyme with specificity against contaminating lactic acid bacteria. Purified endolysins maintain their near-species specificity and can lyse both biofilm and senescent forms of Gram positive bacteria when exposed externally. The expression of these enzymes in yeast will be accomplished via a proprietary methodology that avoids inactivation due to aberrant glycosylation, a major pitfall of heterologous yeast expression systems.