Durham D R, Perry J J
J Bacteriol. 1978 Jun;134(3):837-43. doi: 10.1128/jb.134.3.837-843.1978.
The primary amine dehydrogenase of Pseudomonas putida NP was purified to homogeneity as judged by polyacrylamide gel electrophoresis. Cytochrome c or an artificial electron acceptor was required for amine dehydrogenase activity. The enzyme was nonspecific, readily oxidizing primary monoamines, benzylamine, and tyramine; little or no measurable activity was detected with isoamines, L-ornithine, L-lysine, and certain diamines or polyamines. The pH optima for n-butylamine, benzylamine, and n-propylamine were 7.0, 6.5, and 7.0, respectively. The molecular weight of the enzyme was 112,000 as determined by gel filtration and 95,300 as analyzed by sedimentation equilibrium. Subunit analysis by sodium dodecyl sulfate gel electrophoresis suggested that the enzyme was composed of two nonidentical subunits with molecular weights of 58,000 and 42,000. The absorption spectrum of the purified enzyme was indicative of a hemoprotein, exhibiting absorption maxima at 277, 355, and 408 nm. Reduction with sodium dithionite or amine substrates resulted in absorption maxima at 523 and 552 nm and a shift in the Soret peak to 416 nm. These results suggested that the enzyme is a hemoprotein of the type c cytochrome. There was no evidence that flavins were present.
恶臭假单胞菌NP的伯胺脱氢酶经聚丙烯酰胺凝胶电泳鉴定已纯化至同质。胺脱氢酶活性需要细胞色素c或人工电子受体。该酶具有非特异性，能迅速氧化伯单胺、苄胺和酪胺；对异胺、L-鸟氨酸、L-赖氨酸以及某些二胺或多胺几乎检测不到或没有可测量的活性。正丁胺、苄胺和正丙胺的最适pH分别为7.0、6.5和7.0。通过凝胶过滤测定该酶的分子量为112,000，通过沉降平衡分析为95,300。十二烷基硫酸钠凝胶电泳的亚基分析表明该酶由两个不同的亚基组成，分子量分别为58,000和42,000。纯化酶的吸收光谱表明它是一种血蛋白，在277、355和408nm处有吸收最大值。用连二亚硫酸钠或胺底物还原会导致在523和552nm处出现吸收最大值，并且Soret峰移至416nm。这些结果表明该酶是c型细胞色素类的血蛋白。没有证据表明存在黄素。
