Różalska Małgorzata, Derczyńska Anna, Maszewska Agnieszka
Department of Pharmaceutical Microbiology, Medical University of Lodz, Łódź, Poland.
Department of Immunobiology of Bacteria, Faculty of Biology and Environmental Protection, University of Lodz, Łódź, Poland.
Acta Biochim Pol. 2015;62(4):757-64. doi: 10.18388/abp.2015_1126. Epub 2015 Dec 1.
A total of 104 coagulase negative staphylococci, belonging to S. capitis, S. hominis, S. haemolyticus and S. warneri, originating from the collection of the Department of Pharmaceutical Microbiology (ZMF), Medical University of Lodz, Poland, were tested for their synergistic hemolytic activity. 83% of strains produced δ-hemolysin, however, the percentage of positive strains of S. haemolyticus, S. warneri, S. capitis and S. hominis was different - 98%, 78%, 75% and 68%, respectively. Highly pure hemolysins were obtained from culture supernatants by protein precipitation with ammonium sulphate (0-70% of saturation) and extraction by using a mixture of organic solvents. The purity and molecular mass of hemolysins was determined by TRIS/Tricine PAGE. All CoNS hemolysins were small peptides with a molar mass of about 3.5 kDa; they possessed cytotoxic activity against the line of human foreskin fibroblasts ATCC Hs27 and lysed red cells from different mammalian species, however, the highest activity was observed when guinea pig, dog and human red blood cells were used. The cytotoxic effect on fibroblasts occurred within 30 minutes. The S. cohnii ssp. urealyticus strain was used as a control. The antimicrobial activity was examined using hemolysins of S. capitis, S. hominis, S. cohnii ssp. cohnii and S. cohnii ssp. urealyticus. Hemolysins of the two S. cohnii subspecies did not demonstrate antimicrobial activity. Cytolysins of S. capitis and S. hominis had a very narrow spectrum of action; out of 37 examined strains, the growth of only Micrococcus luteus, Corynebacterium diphtheriae and Pasteurella multocida was inhibited.
对来自波兰罗兹医科大学药物微生物学系（ZMF）菌种保藏库的104株凝固酶阴性葡萄球菌（分别属于头状葡萄球菌、人葡萄球菌、溶血葡萄球菌和沃氏葡萄球菌）进行了协同溶血活性检测。83%的菌株产生δ-溶血素，然而，溶血葡萄球菌、沃氏葡萄球菌、头状葡萄球菌和人葡萄球菌的阳性菌株百分比不同，分别为98%、78%、75%和68%。通过硫酸铵（饱和度0 - 70%）蛋白沉淀和使用有机溶剂混合物萃取，从培养上清液中获得了高纯度的溶血素。溶血素的纯度和分子量通过TRIS/三羟甲基甘氨酸聚丙烯酰胺凝胶电泳测定。所有凝固酶阴性葡萄球菌溶血素均为小肽，摩尔质量约为3.5 kDa；它们对人包皮成纤维细胞系ATCC Hs27具有细胞毒性活性，并能溶解不同哺乳动物物种的红细胞，然而，当使用豚鼠、狗和人的红细胞时观察到最高活性。对成纤维细胞的细胞毒性作用在30分钟内出现。解脲棒状杆菌亚种菌株用作对照。使用头状葡萄球菌、人葡萄球菌、科氏棒状杆菌亚种科氏亚种和解脲棒状杆菌亚种的溶血素检测抗菌活性。两种科氏棒状杆菌亚种的溶血素均未显示抗菌活性。头状葡萄球菌和人葡萄球菌的细胞溶素作用谱非常窄；在37株受试菌株中，仅藤黄微球菌、白喉棒状杆菌和多杀巴斯德菌的生长受到抑制。
