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The goal of this award is to develop a biopreservative ingredient that extends the shelf life food products, starting with soft cheeses such as cottage cheese. Dairy products are part of a healthy diet for ~80% of Americans, providing essential protein, calcium, and vitamins. Dairy foods are highly susceptible to microbial spoilage, but chemical preservatives used in dairy foods are increasingly restricted, due to safety concerns. Thus, dairy manufacturers urgently need effective natural alternatives, known as "biopreservatives", which use microbial fermentation to naturally inhibit spoilage organism growth. Whereas chemical preservatives are single molecules with one antimicrobial mechanism of action (MoA), our product combines multiple MoA through rationally selected microbial communities, or "SupercultureTM ingredients" (SCI). Our USDA SBIR PhI research used computational methods and high-throughput screening to identify a novel lead antifungal ingredient. Preliminary Phase I characterization demonstrated that prototype performance significantly exceeds in-market leading biopreservative products and that ingredient production could be scaled up to pilot fermentor volumes. In this Phase II award, we will build on these exciting results by optimizing performance and production, to verify performance at commercial scales, enabling broad commercialization of this high performance biopreservative ingredient, and ultimately reducing spoilage of dairy products.To achieve this goal, we will pursue the following technical objectives (TOs):TO1: Optimize lead ingredient from PhI research and establish economic bioprocess for upstream and downstream manufacturing at commercial scales. Validate one SCI retains similar or better activity with locked-in upstream and downstream processes: production at 10L pilot scales and following spray drying.TO2: Optimize biopreservative performance via focused screening and iteration. Identify at least 3 new lead ingredients which provide 25% improvement in growth inhibition across ≥50% of high priority dairy spoilage organisms tested (ie 15+ strains) compared to current biopreservative solutions.TO3: Test efficacy biopreservative product in realistic dairy matrices. Validate PhI lead in preliminary pilot plant testing at the outset of PhII work, to inform baseline performance in real-world conditions. Internal benchtop application testing in cottage cheese matrix assay to select lead PhII SCI for advancement to 2nd round customer pilot plant trial. Demonstrate PhII lead hit SCI efficacy in full scale commercial pilot in customer plant: target >25% shelf life extension in accelerated shelf life assay. Validate lead SCI in at least one additional soft cheese matrix (>25% shelf life extension in challenge assay compared to current solution)

Sheth, R.
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