The structure and function of peptides are affected by the changes of environmental factors, including temperature, pH value and ionic strength. Sugar groups can form covalent bonds with amino acid side chain groups of peptides to undergo structural modification, and then the glycosylation modification can effectively improve the stability and biological activity of peptide molecules. In this review, the preparation methods and structural characterization strategies of glycosylated peptides were summarized, and the recent research progress on their physiological activities were highlighted. The challenges and future research directions of glycosylated peptides were also discussed.