ZOLLI Z, SANCLEMENTE C L
J Bacteriol. 1963 Sep;86(3):527-35. doi: 10.1128/jb.86.3.527-535.1963.
Zolli, Zeno, Jr. (Michigan State University, East Lansing), and Charles L. San Clemente. Purification and characterization of staphylocoagulase. J. Bacteriol. 86:527-535. 1963.-Separation and extreme purification of coagulase from Staphylococcus aureus strain 70 was achieved by using three cycles of dialysis in ethanol-water mixtures under controlled conditions, followed by molecular sieving through a column of Sephadex G-200. By manipulation of five variables (pH, ionic strength, temperature, protein, and ethanol concentration), the final preparation showed an approximate 3700-fold increase in activity per mg of protein. The successfully isolated coagulase containing 15.0% nitrogen was characterized serologically and chemically. By use of agar diffusion techniques, one zone of precipitation was obtained with the highly purified material. Additional confirmation of purity was evidenced by the appearance of a single peak with cellulose acetate paper electrophoresis with a barbital buffer at pH 8.6. Progressive and eventual elimination of carbohydrate, deoxyribonuclease, lipase, and phosphatase was observed through the four stages of purification. Temperature studies showed that the stability of each fraction was inversely related to its purity.
小泽诺·佐利（密歇根州立大学，东兰辛）和查尔斯·L·圣克莱门特。葡萄球菌凝固酶的纯化与特性。《细菌学杂志》86:527 - 535。1963年。——通过在受控条件下于乙醇 - 水混合物中进行三轮透析，随后通过Sephadex G - 200柱进行分子筛分离，实现了从金黄色葡萄球菌70株中分离并高度纯化凝固酶。通过对五个变量（pH、离子强度、温度、蛋白质和乙醇浓度）的操控，最终制剂每毫克蛋白质的活性显示出约3700倍的增长。成功分离出的含15.0%氮的凝固酶进行了血清学和化学特性分析。通过琼脂扩散技术，用高度纯化的物质获得了一个沉淀区。在pH 8.6的巴比妥缓冲液中进行醋酸纤维素纸电泳时出现单峰，进一步证实了其纯度。在纯化的四个阶段中观察到碳水化合物、脱氧核糖核酸酶、脂肪酶和磷酸酶逐渐并最终被去除。温度研究表明，各组分的稳定性与其纯度呈负相关。
