PURIFICATION AND PROPERTIES OF STAPHYLOCOCCAL DELTA HEMOLYSIN
PURIFICATION AND PROPERTIES OF STAPHYLOCOCCAL DELTA HEMOLYSIN
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DOI：
10.1128/iai.3.3.449-465.1971
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发表时间：
1971-01-01
影响因子：
3.1
通讯作者：
BERNHEIMER, AW
中科院分区：
文献类型：
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作者：
KREGER, AS;KIM, KS;BERNHEIMER, AW
通过羟基磷灰石的吸附和选择性洗脱，然后用水彻底透析，通过聚乙烯吡咯烷酮或聚乙二醇20,000透析浓缩，最后进行水透析，获得大量（每升培养物上清液200毫克）高度纯化的葡萄球菌可溶性δ溶血素。制剂中未检测到碳水化合物、磷或无活性的 280 nm 吸收材料；然而，通过密度梯度离心、凝胶过滤、分析超速离心、羧甲基纤维素色谱、聚丙烯酰胺圆盘凝胶电泳、等电聚焦和电子显微镜的分析表明，溶素是分子异质的。该制剂含有酸性纤维溶素（S20，wof 11.9）和碱性溶素组分，该碱性溶素组分由不同尺寸的颗粒聚集体组成，最大S20，w约为4.9。制剂中未检测到其他葡萄球菌产物。该溶素对许多动物物种的红细胞具有活性，并与葡萄球菌β溶血素对绵羊红细胞具有协同作用。溶于氯仿-甲醇(2:1)，被各种磷脂、正常血清和蛋白水解酶灭活，但对热灭活有部分抵抗力。活性不受 Ca2+、Mg2+、柠檬酸盐、乙二胺四乙酸或半胱氨酸的影响。溶素制剂还破坏细菌原生质体和原生质球、红细胞膜、溶酶体和脂质球，对某些细菌具有生长抑制作用，并使蛋黄琼脂澄清。大量产生兔子和豚鼠皮肤坏死。小鼠和豚鼠的最小静脉注射致死剂量分别约为 110 和 30 mg/kg。
Large amounts (200 mg per liter of culture supernatant fluid) of highly purified staphylococcal soluble delta hemolysin were obtained by adsorption to and selective elution from hydroxyapatite followed by exhaustive dialysis against water, concentration by polyvinylpyrrolidone or polyethylene glycol 20,000 dialysis, and a final water dialysis. No carbohydrate, phosphorus, or inactive 280-nm absorbing material was detected in the preparation; however, analysis by density gradient centrifugation, gel filtration, analytical ultracentrifugation, carboxymethyl cellulose chromatography, polyacrylamide disc gel electrophoresis, isoelectric focusing, and electron microscopy revealed that the lysin was molecularly heterogeneous. The preparation contained an acidic fibrous lysin (S20,wof 11.9) and a basic lysin component composed of a population of granular aggregates of various sizes, with a maximumS20,wof approximately 4.9. No other staphylococcal products were detected in the preparation. The lysin was active against erythrocytes from many animal species and acted synergistically with staphylococcal beta hemolysin against sheep erythrocytes. It was soluble in chloroform-methanol (2:1), was inactivated by various phospholipids, normal sera, and proteolytic enzymes, but was partially resistant to heat inactivation. Activity was not affected by Ca2+, Mg2+, citrate, ethylenediaminetetraacetic acid, or cysteine. The lysin preparation also disrupted bacterial protoplasts and spheroplasts, erythrocyte membranes, lysosomes, and lipid spherules, was growth-inhibitory for certain bacteria, and clarified egg yolk-agar. Large amounts produced dermonecrosis in rabbits and guinea pigs. The minimum lethal intravenous dose for mice and guinea pigs was approximately 110 and 30 mg/kg, respectively.