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编码链霉菌溶血素(S-溶血素)的 SCO1782 基因在变铅青链霉菌 M145 中的功能分析。

Functional analysis of the gene SCO1782 encoding Streptomyces hemolysin (S-hemolysin) in Streptomyces coelicolor M145.

机构信息

Department of Microbial Engineering, College of Engineering, Konkuk University, 1 Hwayang-dong, Gwangjin-gu, Seoul 143-701, Republic of Korea.

出版信息

Toxicon. 2013 Sep;71:159-65. doi: 10.1016/j.toxicon.2013.05.023. Epub 2013 Jun 4.

Abstract

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.

摘要

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。

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