Biosynthesis of Nondigestible Hetero-oligosaccharides by MelA α-Galactosidase and Glucansucrase E81
學生姓名:
林敏桓
指導教授:
方翠筠
學 期:
113上
摘 要:
Bioactive oligosaccharides are important compounds for development of functional foods and ingredients. In this study, lactose derivative hetero-oligosaccharides(HOS) were produced by acceptor reaction of glucansucrase E81 with whey. The synthesis of oligosaccharides was monitored by Thin Layer Chromatography (TLC) analysis followed by 1H and 13C Nuclear Magnetic Resonance (NMR) spectroscopy analysis which demonstrated the glycosylation of lactose with α-(1 → 2), α-(1 → 3) and α-(1 → 4) linkages. Liquid chromatography–mass spectrometry (LC-MS) analysis demonstrated the formation of DP 7 oligosaccharides from whey. This work describes the high capacity of MelA α-galactosidase from Lactobacillus plantarum WCFS1 to transfer galactosyl residues from melibiose to the C6-hydroxyl group of disaccharide-acceptors with β-linkages (lactulose, lactose, and cellobiose) or α-linkages (isomaltulose and isomaltose) to produce novel galactose-containing HOS. A comprehensive NMR characterization of the transfer products derived from melibiose:lactulose reaction mixtures revealed the biosynthesis of α-d-galactopyranosyl-(1 → 6)-β-d-galactopyranosyl-(1 → 4)-β-d-fructose as the main component as well as the presence of α-d-galactopyranosyl-(1 → 3)-β-d-galactopyranosyl-(1 → 4)-β-d-fructose and α-d-galactopyranosyl-(1 → 6)-α-d-galactopyranosyl-(1 → 6)-β-d-galactopyranosyl-(1 → 4)-β-d-fructose. Melibiose-derived α-galactooligosaccharides (α-GOS), manninotriose and verbascotetraose, were also simultaneously synthesized. The synthesized oligosaccharides were not digested under in vitro human gastric juice and α-amylase conditions. The prebiotic effect of lactose derivative hetero-oligosaccharides was tested and no growth in pathogenic strains was observed whereas certain probiotic strains utilised lactose derivative hetero-oligosaccharides as Carbon source and their growth was promoted.
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