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Oxidation–reduction titration of recombinant WT GWD (○) and C1008S (•) at pH 7.9. Oxidation–reduction equilibration was carried out for 2 h at 25°C and at ambient potentials (Eh) defined by different ratios of reduced: oxidized DTT. Aliquots were withdrawn and assayed for GWD activity as described in Materials and Methods. The solid curve was calculated by fitting the data to the Nernst equation.
