SBIR Phase II: Improvement of the Biofuel Fermentation Process by the Phage-mediated Reduction of Contaminating Lactic Acid Bacteria

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This Small Business Innovation Research Phase II project is focused on improving biofuel fermentation processes by developing phage products designed to inhibit bacteria that reduce biofuel fermentation efficiencies. This product will improve biomass conversion efficiencies in the biorefinery. One of the most significant challenges to commercial biofuel fermentation is the presence of lactic acid bacteria (LAB) that compete for the feedstock, produce undesirable organic acids, and inhibit the growth of the fermentative microorganism. Antibiotics are commonly applied to control LAB, which may lead to the emergence of antibiotic resistant strains. Furthermore, antibiotic residues in distillers grains can lower the value of this important ethanol co-product, further weakening the economics of biofuel production. Despite antibiotic applications, most facilities are still impacted by LAB-associated fermentation upsets, reducing ethanol yields and increasing costs. Ecolyse is proposing to develop an entirely new approach for controlling LAB during fuel ethanol fermentation, based on LAB terminating phage formulations. Phages are natural, highly host-specific bacteriolytic agents. Ecolyse is pioneering the development of phage products designed to mitigate bacterial problems during industrial activities. The dynamics of LAB contamination during biofuel fermentation is particularly well suited for phage-based mitigation.The broader impact/commercial potential of this project is to develop a product that not only improves biofuel ethanol fermentation efficiencies but will also reduces the non-medical use of antibiotics. Current control practices for LAB contamination in fermentation facilities include rigorous clean in place policies, adjusting physical parameters, and through the application of chemical biocides and antibiotics. Antibiotics are often the most effective control measure. Regulations designed to reduce antibiotic use fail to encourage viable alternatives. Phage formulations could potentially fill this void for a myriad of industrial applications, including biofuel fermentation. While phage products are being developed for medical and agricultural purposes, Ecolyse is unique in seeking industrial targets. An advantage that phage share with antibiotics over chemical biocides is capacity to specifically kill target bacteria without interacting other microorganisms, including the fermentative yeast. In contrast, chemical biocides are much less selective and doses effective against bacteria may adversely modulate yeast growth. Thus, the innovative application of phage to control LAB in the fuel ethanol fermentation industry will lead to both immediate economic and long-term socio/economic impacts. The phage products will be marketed to the almost 200 ethanol fermentation facilities in the U.S. alone.