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Reverse-phase HPLC analysis of hygromycin A (HA) reduction by Hyg26. (A) Control reaction of HA and NADH without Hyg26; (B) reaction of HA and NADH after incubation with Hyg26, showing the formation of DHHA. An additional peak (11.1 min) eluting earlier than HA is the C-4″ epimer of HA and not a substrate for Hyg26. mAU, milli-absorbance units.
