Crystallization and Properties of Amine Dehydrogenase from Pseudomonas sp.
Crystallization and Properties of Amine Dehydrogenase from Pseudomonas sp.
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假单胞菌胺脱氢酶的结晶和性质。
DOI：
10.1271/bbb1961.52.2255
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发表时间：
1988
期刊：
影响因子：
--
通讯作者：
M. Ameyama
中科院分区：
文献类型：
--
作者：
E. Shinagawa;K. Matsushita;K. Nakashima;O. Adachi;M. Ameyama
从假单胞菌的无细胞提取物中纯化并结晶出胺脱氢酶，通过富集技术从土壤中分离。该结晶酶在聚丙烯酰胺凝胶电泳上产生单一条带，通过Sephadex柱上的凝胶过滤估计该酶的分子量为100，000。在SDS-凝胶电泳中，该酶解离成两个不同的亚基，分子量分别为60，000（脱氢酶）和39，000（细胞色素c）。该酶的吸收光谱在550 nm、524 nm、411 nm和280 nm处显示最大吸收，在350 nm附近显示宽肩，表明该酶是作为一种酶-细胞色素c复合物纯化的。脱氢酶的辅基被鉴定为共价结合的吡咯喹啉醌。该酶对脂肪族单胺、脂肪族二胺、芳香族胺和多胺具有广泛的底物特异性。
An amine dehydrogenase was purified and crystallized from the cell free extract of a Pseudomonas sp., isolated from soil by means of the enrichment technique. The crystalline enzyme gave a single band on polyacrylamide gel electrophoresis and the molecular weight of the enzyme was estimated to be 100,000 by gel filtration on a Sephadex column. Upon SDS-gel electrophoresis, the enzyme was dissociated into two nonidentical subunits having molecular weights of 60,000 (dehydrogenase) and 39,000 (cytochrome c). The absorption spectrum of the enzyme showed absorption maxima at 550 nm, 524 nm, 411 nm and 280 nm, and a broad shoulder at around 350 nm, indicating that the enzyme was purified as a dehydrogenase-cytochrome c complex. The prosthetic group of the dehydrogenase was identified as covalently bound pyrroloquinoline quinone. The enzyme showed a broad substrate specificity toward various amines including aliphatic monoamines, aliphatic diamines, aromatic amines and polyamines.