Investigation of the structure and digestibility of fluorescently labeled carbohydrates using glycoside hydrolases

Abstract

Developing alternatives to the use of antimicrobials is essential to maintaining the sustainability of animal production and lowering the prevalence of antimicrobial resistance. Prebiotics have not been readily adopted due to limited knowledge of the mechanisms driving prebiotic-bacteria-host interactions. Presented here are preliminary insights into the utility of three prebiotic compounds: neoagarooligosaccharides (NAOS) mannooligosaccharides (MOS), and pectic oligosaccharides (POS). The activity of three agarases from Bacteroides uniformis NP1: BuGH86 (an endolytic enzyme that produces NAOS from agarose), as well as BuGH2C and BuGH117B (exolytic enzymes that remove 3,6-anhydro-L-galactose from the non-reducing end of NAOS and D-galactose from the non-reducing end of agarooligosaccharides, respectively) were determined. NAOS, MOS and POS were then used as substrates to generate fluorescent probes for evaluating interactions with bacterial enzymes. For NAOS and MOS, the digestion of ANTS-labeled oligosaccharides was similar to that of native versions; however, digestion of ANTS-labeled POS was considerably hindered.