N-acetylglucosamine (GlcNAc), a glycan precursor, has essential roles in bacterial survival. TheGlcNAc analog, N-azidoacetylglucosamine (GlcNAz) is a widely used glycan probe. Metabolic oligosaccharide engineering (MOE) employing NahK GlcNAc-1-kinase enables UDP-GlcNAz production and incorporation into the peptidoglycan of E. coli. In this work, the mechanism of GlcNAz import in E. coli was investigated using knockout mutant screens. Our results lead to a model for GlcNAz import, consisting of GlcNAz-6-phosphate import by the MurP-Crr phosphotransferase system, induction of transcriptional regulator NagC, and upregulation of GlcNAz import by galactose permease GalP. Additionally, the scope of this MOE strategy was explored toward novel GlcNAz derivatives and N-azidoacetylmannosamine (ManNAz). This revealed GlmU as a bottleneck to incorporating bulkier GlcNAz analogs while hinting at unexplored amino sugar metabolic pathways. This work sheds light on the promiscuity of bacterial sugar transporters and can inform the development of novel cell-permeable sugar probes and MOE strategies.