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

这个小型企业创新研究第二阶段项目的重点是通过开发旨在抑制降低生物燃料发酵效率的细菌的噬菌体产品来改善生物燃料发酵过程。该产品将提高生物炼制中的生物质转化效率。商业生物燃料发酵的最显著挑战之一是存在乳酸菌（LAB），其竞争原料，产生不期望的有机酸，并抑制发酵微生物的生长。抗生素通常用于控制LAB，这可能导致抗生素耐药菌株的出现。此外，酒糟中的抗生素残留物会降低这一重要乙醇副产品的价值，进一步削弱生物燃料生产的经济性。尽管抗生素的应用，大多数设施仍然受到实验室相关的发酵干扰，降低乙醇产量和增加成本。Ecolyse提出开发一种全新的方法来控制燃料乙醇发酵过程中的LAB，基于LAB终止噬菌体配方。噬菌体是天然的、高度宿主特异性的溶菌剂。Ecolyse是开发旨在减轻工业活动中细菌问题的噬菌体产品的先驱。生物燃料发酵过程中LAB污染的动态特别适合于基于噬菌体的缓解。该项目更广泛的影响/商业潜力是开发一种产品，不仅提高生物燃料乙醇发酵效率，还将减少抗生素的非医疗用途。目前发酵设施中LAB污染的控制实践包括严格的就地清洁政策、调整物理参数以及通过应用化学杀生物剂和抗生素。抗生素通常是最有效的控制措施。旨在减少抗生素使用的法规未能鼓励可行的替代品。噬菌体制剂可能填补这一空白，用于无数的工业应用，包括生物燃料发酵。虽然噬菌体产品正在开发用于医疗和农业目的，但Ecolyse在寻求工业目标方面是独一无二的。与化学杀生物剂相比，噬菌体与抗生素共有的优点是能够特异性地杀死靶细菌而不与其他微生物（包括发酵酵母）相互作用。相比之下，化学杀生物剂的选择性要低得多，并且对细菌有效的剂量可能不利地调节酵母生长。因此，噬菌体控制LAB在燃料乙醇发酵工业中的创新应用将导致直接的经济和长期的社会/经济影响。噬菌体产品将销售给美国近200个乙醇发酵设施。

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.