Glycosylation is one of the most important post-modification processes of small molecules and enables the parent molecule to have increased solubility, stability, and bioactivity. Enzyme-based glycosylation has achieved significant progress due to advances in protein engineering, DNA recombinant techniques, exploitation of biosynthetic gene clusters of natural products, and computer-based modeling programs. Our report summarizes glycosylation data that have been published within the past five years to provide an overall review of current progress. We also present the future trends and perspectives for glycosylation.