Chung J G
Department of Medicine, China Medical College, Taichung 400, Taiwan, Republic of China.
Curr Microbiol. 1998 Jul;37(1):70-3. doi: 10.1007/s002849900341.
N-acetyltransferase from Aeromonas hydrophilia was purified by ultrafiltration, DEAE-Sephacel, gel filtration chromatography on Sephadex G-100, and DEAE-5pw on high performance liquid chromatography, as judged by sodium dodecyl sulfate-polyacrylamine gel electrophoresis (SDS-PAGE) on a 12.% (wt/vol) slab gel. The enzyme had a molecular mass 44.9 kDa. The purified enzyme was thermostable at 37 degrees C for 1 h with a half-life 28 min at 37 degrees C, and displayed optimum activity at 37 degrees C and pH 7.0. The Km and Vmax values for 2-aminofluorene were determined to be 0. 896 mM and 2.456 nmol/min/mg protein, respectively. Among a series of divalent cations and salts, Zn2+, Ca2+, and Fe2+ were demonstrated to be the most potent inhibitors.
通过超滤、DEAE-葡聚糖凝胶、Sephadex G-100凝胶过滤色谱以及高效液相色谱上的DEAE-5pw对嗜水气单胞菌的N-乙酰转移酶进行纯化，这是根据在12%（重量/体积）平板凝胶上进行的十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）判断的。该酶的分子量为44.9 kDa。纯化后的酶在37℃下1小时内具有热稳定性，在37℃下的半衰期为28分钟，并且在37℃和pH 7.0时表现出最佳活性。2-氨基芴的Km和Vmax值分别测定为0.896 mM和2.456 nmol/分钟/毫克蛋白质。在一系列二价阳离子和盐中，Zn2+、Ca2+和Fe2+被证明是最有效的抑制剂。
