Purification and properties of an amine dehydrogenase from Pseudomonas K95 grown on 1,12-diaminododecane (DAD).

Abstract

The cell-free extract of Pseudomonas K95 gwown on 1,12-diaminododecane (DAD) could oxidize all tested alkyl diamines, from C4 to C12. The cell-free extract contained an amine dehydrogenase. The amine dehydrogenase was purified 1700-fold to homogeneity from Pseudomonas K95 grown on DAD as the sole carbon source. The molecular weight of the enzyme was found to be 56,000, on gel filtration, and 47,000, on sodium dodecyl sulfate gel electrophoresis; no evidence was obtained for subunits. The enzyme is able to utilize only phenazine methosulphate as an electron acceptor. The enzyme is markedly nonspecific, readily oxidizing both short and long chain primary monoamines and diamines, polyamines, l-noradrenaline, histamine, benzylamine and di-n-hexylamine. The enzyme was inhibited by the carbonyl reagents, semicarbazide and isoniazid. The optimum pH for the oxidation of DAD was 7.0 and that for 12-aminododecanoic acid (ADA) was 8.0. The Km value for DAD at pH 7.0 was 3 μ and that for ADA was 33μm. The difference ...