Hewlett E, Wolff J
J Bacteriol. 1976 Aug;127(2):890-8. doi: 10.1128/jb.127.2.890-898.1976.
Culture medium of exponentially growing Bordetella pertussis (strain 114) contains significant quantities of soluble (100,000 X g for 1 h) adenylate cyclase. The enzyme was purified by chromatography on diethylaminoethyl-cellulose and Sephadex G-200. The purest material yielded a single band on sodium dodecyl sulfate-disc gel electrophoresis. It is heat labile, has a temperature optimum of 30 degrees C, a pH optimum of pH 7 to 8, and a Km for adenosine 5'-triphosphate of 0.4 mM, and requires Mg2+ for maximum activity. The molecular weight, by sodium dodecyl sulfate-disc gel electrophoresis and sucrose density gradient, is approximately 70,000. The enzyme is markedly inhibited by fluoride and weakly inhibited by monovalent salts, but its activity is not altered by alpha-keto acids of nonsubstrate nucleoside triphosphates. Thus, but its presence in the culture supernatant, its smaller molecular weight, and its insensitivity to alpha-keto acids and nucleotides, this enzyme differs from the bacterial adenylate cyclases previously described.
处于指数生长期的百日咳博德特氏菌（菌株114）的培养基含有大量可溶性（100,000×g离心1小时）腺苷酸环化酶。该酶通过在二乙氨基乙基纤维素和葡聚糖G - 200上进行层析纯化。最纯的物质在十二烷基硫酸钠圆盘凝胶电泳上产生一条带。它对热不稳定，最适温度为30℃，最适pH为7至8，对腺苷5'-三磷酸的Km值为0.4 mM，并且需要Mg2+以达到最大活性。通过十二烷基硫酸钠圆盘凝胶电泳和蔗糖密度梯度法测定，其分子量约为70,000。该酶受到氟化物的显著抑制，受到单价盐的微弱抑制，但其活性不受非底物核苷三磷酸的α-酮酸的影响。因此，鉴于其存在于培养上清液中、分子量较小以及对α-酮酸和核苷酸不敏感，这种酶与先前描述的细菌腺苷酸环化酶不同。
