Peptidoglycan Hydrolase Antimicrobials for Eradicating Lactobacilli that Contaminate and Reduce Ethanol Yields in Biofuel Fermentation

摘要

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.

主权项

1. A method of controlling bacterial contamination in ethanol fermentation comprising: introducing a fermentation mixture comprising yeast S. cerevisiae into a vessel used during ethanol production; introducing a recombinant bacteriophage lytic endolysin as an active agent to the vessel used during ethanol production, and fermenting the fermentation mixture by said yeast to produce ethanol; wherein the recombinant endolysin is an antimicrobial agent having exolytic activity for at least one targeted Gram positive lactic acid bacterial species in an amount effective to at least reduce the levels of growth of and colonization by at least one targeted lactic acid bacterial species present in the vessel during ethanol production without inhibiting the growth of said yeast in the vessel thereby resulting in increased ethanol production from the fermentation mixture.