Peptidoglycan hydrolase antimicrobials for eradicating lactobacilli that contaminate and reduce ethanol yields in biofuel fermentation

US9068204B2 · kind B2 · utility

Assignee

• The United States of America, as represented by the Secretary of Agriculture · US

Inventors

• David M. Donovan · Baltimore, US
• Dwayne Roach · Bakersfield, US
• Piyum A. Khatibi · Dunlap, US
• Kenneth M. Bischoff · Morton, US
• Stephen R. Hughes · Peoria, US

Key dates

• Filing date: Mar 15, 2013
• Grant date: Jun 30, 2015
• Expiry date: Aug 2, 2033

Classification

• Technology area (CPC Y): Emerging Cross-Sectional Technologies
• CPC primary: Y02E50/10
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

Ethanol losses due to bacterial contamination in fermentation cultures weakens the economics of biofuel production. Lactobacillus species are the predominant contaminant. Bacteriophage lytic enzymes are peptidoglycan hydrolases which degrade Gram positive cell walls when exposed externally and are a novel source of antimicrobials. The streptococcal phage λSA2 endolysin construct demonstrated strong lytic activity towards 17 of 22 strains of lactobacilli, staphylococci or streptococci maintaining optimal specific activity under fermentation conditions toward L. fermentum substrates. Lactobacillus bacteriophage endolysin constructs LysA, LysA2 and LysgaY showed exolytic activity towards ˜60% of the lactobacilli tested including four L. fermentum isolates from fuel ethanol fermentations. Presence of ethanol (≦5%) did not affect lytic activity. Lysins were able to reduce both L. fermentum and L. reuteri contaminants in mock fermentations of corn fiber hydrolysates. Recombinant LysA and λSa2 expressed in the yeast Saccharomyces cerevisiae are functional; LysA was shown to reduce lactobacilli in experimentally infected fermentations.