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Two-step enzymatic reaction cycle for the production of 2,7-anhydro-Neu5Ac from Neu5Ac. A) Step A, OP2E α2–3-sialylation system containing NmCSS and PmST1_M144D for the formation of Neu5Acα2–3LacβProNHCbz (5) or Neu5Acα2–3LacβNHCbz (7) from LacβProNHCbz (3) or LacβNHCbz (6) in the presence of Neu5Ac and CTP; and B) Step B, SpNanB-catalyzed formation of 2,7-anhydro-Neu5Ac (1) from Neu5Acα2–3LacβProNHCbz (5) or Neu5Acα2–3LacβNHCbz (7) produced in Step A.
