葡萄球菌弹性蛋白酶的产生与检测
Production and detection of staphylococcal elastase.
作者信息
Hartman D P, Murphy R A
出版信息
Infect Immun. 1977 Jan;15(1):59-65. doi: 10.1128/iai.15.1.59-65.1977.
The optimum conditions were determined for the production and detection of staphylococcal elastase from Staphylococcus epidermidis. Optimum production and recovery took place when the dialysis membrane technique was utilized with brain heart infusion agar incubated for 44 h under 15% CO2 and harvested in physiological saline. Near-optimal production took place by 28 h, and this was found more useful for routine use. Incorporation of elastin into the culture medium or the inoculum did not result in higher levels of elastase production. The detection system consisted of 0.25% particulate elastin suspended in a pH 7.0, 0.05 M tris(hydroxymethyl)aminomethane-hydrochloride-buffered solidified plated medium. Crude undialyzed elastase was best detected when the medium contained either agarose and 10(-3) M calcium or purified agar without additional additives. Crude dialyzed elastase was best detected when the medium contained agarose and 10(-3) M disodium ethylenediaminetetraacetic acid.
确定了表皮葡萄球菌产葡萄球菌弹性蛋白酶及检测该酶的最佳条件。当采用透析膜技术，将脑心浸液琼脂在15%二氧化碳条件下孵育44小时，然后在生理盐水中收获时，可实现最佳产量和回收率。28小时时产量接近最佳，且发现这对常规使用更有用。在培养基或接种物中加入弹性蛋白并不会导致更高水平的弹性蛋白酶产生。检测系统由悬浮于pH 7.0、0.05 M三（羟甲基）氨基甲烷 - 盐酸缓冲凝固平板培养基中的0.25%颗粒状弹性蛋白组成。当培养基含有琼脂糖和10⁻³ M钙或不含其他添加剂的纯化琼脂时，未透析的粗弹性蛋白酶检测效果最佳。当培养基含有琼脂糖和10⁻³ M乙二胺四乙酸二钠时，透析后的粗弹性蛋白酶检测效果最佳。
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