This review describes the relationship between hetero-oligosaccharides and microorganisms. It is possible to prepare aminosugar nucleotides as donors for hetero-oligosaccharide synthesis with a combination of yeast fermentation and bacterial enzymes, and to use the product to test for a rare human blood group. We have isolated various glycosidases produced by microorganisms, mainly from soil, to elucidate the structure and function of hetero-oligosaccharides. Among them, a mold endoglycosidase was found to have specific transglycosylation activity in addition to hydrolysis activity, and we have used it to synthesize chemo-enzymatically various bioactive glycopeptides by the attachment of a hetero-oligosaccharide to a peptide. We found that lactic acid bacteria bound to a hetero-oligosaccharide on the intestinal tract cell surface in animals. We also analyzed the bifidobacterial hetero-oligosaccharide-hydrolyzing enzymes involved in the degradation of mucin glycoprotein in the host intestinal tract and human milk oligosaccharides, and identified a specific saccharide that acted as a bifidobacteria growth factor.