Enzymatic modification of isolated bovine milk oligosaccharides to mimic human milk oligosaccharides with increased antimicrobial activity

DESCRIPTION provided by applicant Consumption of human milk is associated with reduced incidence and severity of enteric infections leading to reduced morbidity and mortality in infants Human milk oligosaccharides HMO are known to contribute to this outcome through various mechanisms including a decoy mechanism and by increasing the growth of beneficial microbes Increased growth of specific commensals improves the barrier function of the intestinal epithelium and provides spatial competition reducing pathogen colonization While several HMO have been successfully synthesized no large scale source for the majority of known HMO exists Large quantities of HMO like oligosaccharide OS are vital for preclinical testing of their antimicrobial functions Bovine milk contains HMO like OS BMO but they are typically less fucosylated The fucosylation and sialylation of HMO is important for both direct antimicrobial action via the decoy effect and indirect antimicrobial action via commensal growth enhancement Low levels of fucosylation and sialylation result in OS with poor specificity of action for growth promotion During cheese production whey permeate is produced as a waste by product and it contains all of the BMO We can acquire large quantities whey permeate from industrial cheese manufacturers and isolate the BMO However to increase their direct and indirect antimicrobial potency these OS must be modified to increase their sialylation and fucosylation By enzymatic modification we will create a novel class of BMO based OS with increased antimicrobial activities First we will isolate gram to kilogram quantities of OS from bovine milk whey waste streams Next we will modify their structures with fucosylation sialylation or a combination of both Finally we will examine the original and modified OS for antimicrobial actions against several pathogens for decoy action and several commensals for prebiotic specificity Phase I will demonstrate the feasibility of this approach to generate kilogram quantities of HMO like antimicrobial OS for preclinical testing PUBLIC HEALTH RELEVANCE First we will isolate gram to kilogram quantities of oligosaccharides from bovine milk whey waste streams Next we will modify the bovine milk oligosaccharides with fucosylation sialylation or a combination of both Finally we will examine the original and modified oligosaccharides for antimicrobial actions against several pathogens for decoy action and several commensals for prebiotic specificity Phase I will demonstrate the feasibility of this approach to generate kilogram quantities of human milk oligosaccharides like antimicrobial oligosaccharides for preclinical testing