以致病性哈氏弧菌(Vibrio harveyi)腹腔注射布氏石斑鱼(Epinephelus bleekeri)18h后,取布氏石斑鱼头肾制备头肾粗提物,并对该粗提物的抗菌活性进行研究,结果表明:布氏石斑鱼经哈氏弧菌诱导后,其头肾粗提物不仅对金黄色葡萄球菌(Staphyloccocus aureus)等革兰氏阳性菌有良好抑制作用,而且对大肠杆菌D31(Escherichio coli D31)、哈氏弧菌和溶藻弧菌(V.alginolyticus)等革兰氏阴性菌也有良好抑制效果;利用电泳凝胶琼脂糖弥散法检测发现,头肾粗提物中具有多种对大肠杆菌D31有较强作用效果的蛋白类抗菌物质;切胶回收AU—PAGE凝胶上的抗菌组分,利用SDS—PAGE凝胶检测各抗菌活性组分的分子质量大小依次约为:10.9KD,10.0KD,8.1KD,7.2KD,6.3KD,由于这些抗菌活性成分可被蛋白酶破坏,且分子质量均接近或低于10KD,可初步将其认定为抗菌肽。
The pathogenic Vibrio harveyi was intraperitoneal injected to Epinephelus bleekeri, and after 18h, the crude extract was prepared from the head-kidney of E. bleekeri, and the analysis results of the antibacterial activities of the crude head-kidney extract indicated that the crude head-kidney extract have good antibacterial effects on both G^+ bacteria and G^- bacteria. Radial Diffusion Assay (RDA) in Agarose Gels and Acid-urea Polyacrylamide Gel Electrophoresis (AU-PAGE) methods were used to detect and screen the antibacterial materials , and the data showed that the crude head-kidney extracts contain at least 5 kinds of antibacterial materials with good antibacterial effects on Escherichia coli D31, and all the antibacterial materials were obtained after DNA gel extraction from AU-PAGE gel , whose molecular mass were approximately 10.9 KD, 10.0 KD, 8.1 KD, 7.2 KD and 6.3 KD, respectively. Because the antibacterial materials could be destroyed and their molecular mass was close to or lower than 10 KD, they were preliminarily considered as antibacterial peptides.