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Steady-state kinetic curves of KshA1 and KshA5. Shown is the dependence of the initial velocity of NADH consumption by KshA1 on 4-BNC (A) and O2 (B), and by KshA5 on 5α-androstan-3,17-dione (C), ADD (D), 4-BNC (E), and O2 (F). Assays were performed using 100 μm NADH and potassium phosphate (I = 100 mm, pH 7.0) at 22 °C. B, steroid substrate was 50 μm 4-BNC; F, steroid substrate was 5 μm 4-estren-3,17-dione. Curves represent the best fit of the Michaelis-Menten equation using LEONORA.
