Bunyatratchata, ApichayaApichayaBunyatratchataYU-PING HUANGOzturk, GulustanGulustanOzturkCohen, Joshua L.Joshua L.CohenBhattacharya, MrittikaMrittikaBhattacharyaMln De Moura Bell, JulianaJulianaMln De Moura BellBarile, DanielaDanielaBarile2025-08-052025-08-052020-12-230021-85611520-5118https://www.scopus.com/pages/publications/85097879482https://scholars.lib.ntu.edu.tw/handle/123456789/730960N-Glycans are structurally similar to human milk oligosaccharides, the gold standard prebiotics for infants. Bovine milk N-glycans released by endo-β-N-acetylglucosaminidase (EndoBI-1) were shown to have similar prebiotic selectivity as human milk oligosaccharides, explaining the interest for N-glycan recovery for use as prebiotics. Industrial thermal treatments such as high-temperature short-time (HTST) and ultra-high-temperature (UHT) might favor the enzymatic deglycosylation of N-glycans through promoting protein denaturation. We investigated the effects of HTST (72 °C for 15 s) and UHT (135 °C for 3 s) on N-glycan release from bovine colostrum glycoproteins by nonimmobilized and amino-immobilized EndoBI-1. A total of 104 N-glycans including isomers/anomers were identified by high-resolution mass spectrometry. In both EndoBI-1 forms, HTST increased the release of N-glycans; however, the impact of UHT on releasing N-glycans was comparable to the nonthermal treatment. Although the amino-immobilized enzyme similarly released neutral N-glycans as the free form, it released fewer sialylated and fucosylated N-glycans.enfalsebovine milkimmobilized enzymemass spectrometryN-glycansprebioticsjournal article10.1021/acs.jafc.0c059862-s2.0-85097879482