Department of Microbial Engineering, College of Engineering, Konkuk University, 1 Hwayang-dong, Gwangjin-gu, Seoul 143-701, Republic of Korea.
Toxicon. 2013 Sep;71:159-65. doi: 10.1016/j.toxicon.2013.05.023. Epub 2013 Jun 4.
In the process of evaluating the growth of Streptomyces coelicolor on rich media such as blood agar, we found that S. coelicolor a non-pathogenic, well-known antibiotic producer had the ability to grow and produce a prominent hemolytic zone. By comparing the growth with an agarase gene mutant of S. coelicolor, a similar prominent hemolytic zone was found to develop due to the organism's hemolytic activity. After the confirmation of hemolytic activity from S. coelicolor, the genome was searched for hemolysin-coding genes; consequently, SCO1782, SCO2534, and SCO3882 were identified, whose products were annotated as a putative, membrane, and hypothetical proteins, respectively. Functional characterization of all the recombinant proteins expressed in Escherichia coli BL21(DE3) revealed that only SCO1782 exhibited hemolytic activity. This S. coelicolor protein, designated as S-hemolysin, showed sequence similarity toward hemolysins from Brachyspira hyodysenteriae (35%) and Mycobacterium tuberculosis (62%). Recombinant hemolysin exhibited activity against sheep blood erythrocytes and cytolytic activity against human fibroblast cells. Deletion of SCO1782 resulted in complete loss of hemolysin activity in S. coelicolor.
在评估土黄色链霉菌(Streptomyces coelicolor)在富含血液琼脂等培养基上的生长过程中,我们发现这种非致病性、著名的抗生素产生菌具有生长和产生明显溶血圈的能力。通过比较土黄色链霉菌琼脂酶基因突变体的生长情况,发现由于该生物的溶血活性,也会产生类似的明显溶血圈。在确认土黄色链霉菌的溶血活性后,我们在基因组中搜索了编码溶血素的基因;结果鉴定了 SCO1782、SCO2534 和 SCO3882,其产物分别被注释为推定的、膜和假设的蛋白质。在大肠杆菌 BL21(DE3) 中表达的所有重组蛋白的功能表征表明,只有 SCO1782 表现出溶血活性。这种土黄色链霉菌蛋白被命名为 S-溶血素,它与猪痢疾密螺旋体(Brachyspira hyodysenteriae)(35%)和结核分枝杆菌(Mycobacterium tuberculosis)(62%)的溶血素有序列相似性。重组溶血素对绵羊血红细胞具有活性,对人成纤维细胞具有细胞溶解活性。缺失 SCO1782 会导致土黄色链霉菌中溶血素活性完全丧失。