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The draft genome of Staphylococcus warneri TRPF4, a bacteriocin producer with potent activity against the causative agent of Legionnaires' Disease.
3 Biotech ( IF 2.8 ) Pub Date : 2020-05-06 , DOI: 10.1007/s13205-020-02231-3
Fernanda de Souza Freitas 1 , Pedro Marcus Pereira Vidigal 2 , Tatiane de Paula Siqueira 1 , Mariana de Barros 3 , Marcos Rogério Tótola 1
Affiliation  

In this work, we present the draft genome sequence of Staphylococcus warneri strain TRPF4 consisting of 2,634,550 bp with a G + C content of 32.4%. The genome sequence includes 2466 protein-coding genes, 11 rRNAs and 62 tRNAs, in 33 contigs. Applying the Rapid Annotation using Subsystem Technology (RAST) a total of 1322 protein-coding genes were assigned to 393 subsystems. Also, a set of 1286 protein-coding genes with designated functions were assigned to 21 categories in the Cluster of Orthologous Groups (COG) database. Further analysis of BAGEL3 software demonstrated that the TRPF4 genome contains two gene clusters responsible for the synthesis of three bacteriocins, one warnericin RK and two delta-lysins. Besides, a novel delta-lysin of 3.48 kDa was identified for the first time. The three predicted bacteriocins were chemically synthesized and screened for the antimicrobial activity against a range of pathogens, exhibiting a potent and specific antimicrobial activity counter to L. pneumophila, with minimum inhibitory concentrations (MIC) ranging from 1.9 to 7.8 µg mL-1. These results indicate that the strain TRPF4 can produce bacteriocins with anti-Legionella activity. This was verified by the extracting the bacteriocins from the fermentation broth and testing against L. pneumophila. Additionally, the strain TRPF4 exhibited no cytotoxicity in mammalian cell lines. In summary, the genomic sequences and in vitro assays demonstrated the potential application of bacteriocins from S. warneri TRPF4 as a scaffold for further development of drugs against L. pneumophila, the causative agent of Legionnaires' Disease.

中文翻译:

华氏葡萄球菌TRPF4(一种细菌素生产者,对军团病的致病因子具有有效活性)的基因组草案。

在这项工作中,我们提出了由2,634,550 bp组成的金黄色葡萄球菌TRPF4菌株的基因组序列草案,其G + C含量为32.4%。基因组序列包括33个重叠群中的2466个蛋白质编码基因,11个rRNA和62个tRNA。使用子系统技术(RAST)进行快速注释,总共将1322个蛋白质编码基因分配给393个子系统。此外,将一组具有指定功能的1286个蛋白质编码基因分配给直系同源群(COG)数据库中的21个类别。BAGEL3软件的进一步分析表明,TRPF4基因组包含两个基因簇,这些基因簇负责合成三种细菌素,一种warnericin RK和两种delta-lysins。此外,首次发现了一种新型的3.48 kDa的δ-溶素。化学合成了三种预测的细菌素,并筛选了对一系列病原体的抗菌活性,显示出与嗜肺乳杆菌相反的有效和特异性抗菌活性,最小抑菌浓度(MIC)为1.9至7.8 µg mL-1。这些结果表明菌株TRPF4可以产生具有抗军团菌活性的细菌素。通过从发酵液中提取细菌素并测试嗜肺乳杆菌来验证这一点。另外,菌株TRPF4在哺乳动物细胞系中没有表现出细胞毒性。总而言之,基因组序列和体外试验证明了来自华氏链球菌TRPF4的细菌素作为支架进一步开发抗肺炎嗜血杆菌的药物的潜在应用,嗜肺嗜血杆菌是军团病的病原体。
更新日期:2020-05-06
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