PURIFICATION AND CHARACTERIZATION OF MONOAMINE-OXIDASE FROM KLEBSIELLA-AEROGENES

The gene for monoamine oxidase (maoA) from Klebsiella aerogenes W70 ha s been cloned and the enzyme was overproduced in a soluble form. The e nzyme was purified approximately 10-fold to homogeneity. The enzyme ha s a molecular weight of about 79,000, which is identical to the molecu lar weight deduced from the nucleotide sequence of the gene for monoam ine oxidase. The enzyme had maximum activity at pH 6.0 and 50-degrees- C when catalyzing the oxidative deamination of tyramine. The enzyme ca talyzed the deamination of beta-phenylethylamine, dopamine, tryptamine , and octopamine, but not of diamines, polyamines, or amino acids. The enzyme was inhibited by clorgyline, isoniazid, and carbonyl reagents, but not by 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide. The enzyme did not exhibit a typical flavoprotein spectrum, but the enzymatic ac tivity increased linearly with increasing amounts of added copper. The purified enzyme was found to contain 10 g of copper per 79,000 g of t he protein. The enzymological properties and the amino acid sequence o f the enzyme deduced from the nucleotide sequence of the moaA gene are different from those of known tyramine or monoamine oxidases.