Pro-Hyp (PO) accounts for many beneficial biological effects of collagen hydrolysates for skin and bone health. The objective of this study was to conjugate PO with glucosamine (GlcN) to create a novel glycopeptide Pro-Hyp-CONH-GlcN (POGlcN) and then to investigate the potential involvement of multiple transepithelial transport pathways for this glycopeptide. Nuclear magnetic resonance results revealed the amide nature of this glycopeptide with α and β configurations derived from GlcN. This glycopeptide was very resistant to simulated gastrointestinal digestion. Also， it showed a rate of transepithelial transport [permeability coefficient (P) of (2.82 ± 0.15) × 10 cm/s] across the Caco-2 cell monolayer superior to those of parental dipeptide PO and GlcN [P values of (1.45 ± 0.17) × 10 and (1.87 ± 0.15) × 10 cm/s， respectively]. A transport mechanism experiment indicated that the improved transport efficiency of POGlcN is attributed to the introduction of glucose transporters.