Bacterial Monoamine Oxidases |
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Authors: | Hideaki Yamada Takayuki Uwajima Hidehiko Kumagai Masahiro Watanabe Koichi Ogata |
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Institution: | 1. Research Institute for Food Science, Kyoto University, Kyoto;2. Laboratory of Applied Microbiology, Department of Agricultural Chemistry, Kyoto University, Kyoto |
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Abstract: | A procedure for obtaining crystalline preparations of tyramine oxidase of Sarcina lutea has been developed. The procedure included fractionation with ammonium sulfate, treatment with protamine sulfate and separation by column chromatographies on DEAE-cellulose, hydroxylapatite and sephadex G-150. The specific activity of enzyme was increased 5,700~ 6,000-fold through the procedure, over the crude cell extract. Crystals were prepared from solutions of the purified enzyme by adding solid ammonium sulfate. The crystals appeared as minute, highly refractive needles, with a bright yellow color.With the use of crystalline preparations of tyramine oxidase of Sarcina lutea, substrate and inhibitor specificities of the enzyme were investigated. The enzyme oxidized tyramine and dopamine at almost the same rates. Other monoamines, diamines, polyamines and amino acids were not oxidized at all. The oxidation of tyramine proceeded as follows: Tyramine+O2+H2O→p-Hydroxyphenylacetaldehyde +NH3+H2O2. Ammonia and hydrogen peroxide were formed in stoichiometric amounts.The enzyme was not inhibited by carbonyl reagents, such as hydroxylamine, hydrazine, semicarbazide and isoniazid, but was inhibited by p-CMB and iproniazid. |
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