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United States Department of Agriculture

Agricultural Research Service

Research Project: Developing Bactericidal Yeast for Cellulosic Fuel Ethanol Production

Location: Renewable Product Technology Research Unit

Project Number: 5010-41000-135-04
Project Type: Reimbursable

Start Date: Dec 15, 2009
End Date: Jun 14, 2014

Objective:
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.

Approach:
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.

Last Modified: 10/30/2014
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