PURIFICATION AND CHARACTERIZATION OF THE ALKENE MONOOXYGENASE FROM NOCARDIA-CORALLINA B-276

Alkene monooxygenase from the propene utilizer Nocardia corallina B-276 was separated into three components, and all components were purified to homogeneity and their properties were examined, The epoxidase, with a molecular mass of 95kDa, was considered to catalyze the oxidation of the substrate propene to propylene oxide, It consisted of 53- and 35-kDa subunits, which contained approximately 2-mol of non-heme iron per mole of protein. The reductase, molecular mass 40 kDa, was found to contain an FAD and an Fe-2 S-2 cluster, A third protein, which we have called the coupling protein, with a mass of 14 kDa, appears to function as a regulator of activity. The purified AMO system required NADH as an electron donor, and catalyzed alkene epoxidation only, Acetylene, a specific inhibitor for methane monooxygenase, did not inhibit the AMO activity.