ABSTRACTStudies of the microbiome of older individuals has shown greatly reduced numbers of known beneficialbacteria coinciding with a greater presence of potentially detrimental bacterial strains. Not surprisingly, thedecrease in beneficial microbes coincides with a loss of anti-inflammatory activity and genes associated withshort-chain fatty acid production. This age-related decline in the gut microbiome is also an importantdeterminant of age-associated pathological states including neurodegeneration, chronic inflammation, frailty,cognitive decline, type 1 and type 2 diabetes, musculoskeletal degeneration, as well as nonalcoholic fatty liverdisease, cardiovascular disease, and development of certain types of cancer. The need to increase overallconsumption of resistant starch (RS) prebiotics in the elderly population is clear. The difficulty arises inproviding RS in sufficient quantities (20?30g/daily) to these individuals in a form that is easily stored andreadily consumed. In this project, Design-Zyme LLC will 1) Develop a rapid assay for evaluation of prebioticnutrient component efficacy on known human gut microbes, and 2) Optimize a safe, digestion-resistantmolecule specifically targeting beneficial human gut microbiota deficiencies in the elderly. To accomplish this,we will design an assay to quickly and easily evaluate the growth of common strains of beneficial gut bacteriaand then use this assay to screen individual molecular components of a known resistant starch prebioticcompound. At the end of Phase I, we will have developed an easy-to-use assay for determining effectivenessof individual prebiotics. Furthermore, using this assay, we will have identified specific molecules that promotethe proliferation of beneficial gut bacteria, especially those in the aging and elderly populations. During PhaseII of this proposal, we will establish how to best modify and formulate the molecules from Phase I to resistbreakdown in solution while concomitantly allowing for more concentrated solutions to be manufactured. Atthe end of Phase II, we will have established how to protect these molecules from breakdown in a watersolution, allowing for the creation of pre-mixed aqueous beverages capable of retaining their prebioticfunctionality with a minimum shelf-life of six months. Research into prebiotic nutrient compounds remains animportant, emerging field. The techniques and assays developed in this grant proposal will be widelyapplicable to a range of prebiotic compounds and will advance the development of effective, concentratedprebiotics capable of directly modulating the human gut microbiome and improving overall health in the elderlypopulation.