Fig. 1. Engineered biosynthesis for UDP-GalNAc analogues. (A) Biosynthetic pathways. UDP-GalNAc is biosynthesized by GALK2 and AGX1 activities, while modifications often require the enzymes NahK and AGX1F383A. (B) Acceptance of GalNAc as well as azide-tagged analogues by sugar-1-kinases as measured by UPLC-MS in 16 h reactions. Data are individual data points with means + SD from three independent replicates. (C) Acceptance of GalNAc-1-phosphate as well as azide-tagged analogues by AGX1 constructs as assessed by UPLC. GalNAc-1-phosphate analogues were generated in situ by reaction of the corresponding monosaccharides with NahK as shown in (B). Data are individual data points with means + SD from three independent replicates.
