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一株针对表达K2荚膜的高毒力菌株52145的新型裂解性噬菌体的分离、鉴定及其功能解聚酶的识别

Isolation and Characterization of a Novel Lytic Bacteriophage against the K2 Capsule-Expressing Hypervirulent Strain 52145, and Identification of Its Functional Depolymerase.

作者信息

Pertics Botond Zsombor, Cox Alysia, Nyúl Adrienn, Szamek Nóra, Kovács Tamás, Schneider György

机构信息

Department of Medical Microbiology and Immunology, Medical School, University of Pécs, Szigeti st. 12., H-7624 Pécs, Hungary.

Department of Biotechnology, Nanophagetherapy Center, Enviroinvest Corporation, Kertváros st. 2., H-7632 Pécs, Hungary.

出版信息

Microorganisms. 2021 Mar 21;9(3):650. doi: 10.3390/microorganisms9030650.

Abstract

is among the leading bacteria that cause nosocomial infections. The capsule of this Gram-negative bacterium is a dominant virulence factor, with a prominent role in defense and biofilm formation. Bacteriophages, which are specific for one bacterial strain and its capsule type, can evoke the lysis of bacterial cells, aided by polysaccharide depolymerase enzymes. In this study, we isolated and characterized a bacteriophage against the nosocomial 52145 strain with K2 capsular serotype. The phage showed a narrow host range and stable lytic activity, even when exposed to different temperatures or detergents. Preventive effect of the phage in a nasal colonization model was investigated in vivo. Phlyogenetic analysis showed that the newly isolated phage B1 belongs to the genus in family. We identified the location of the capsule depolymerase gene of the new phage, which was amplified, cloned, expressed, and purified. The efficacy of the recombinant B1dep depolymerase was tested by spotting on strains and it was confirmed that the extract lowers the thickness of the bacterium lawn as it degrades the protective capsule on bacterial cells. As strains possessing the K2 serotype have epidemiological importance, the B1 phage and its depolymerase are promising candidates for use as possible antimicrobial agents.

摘要

是引起医院感染的主要细菌之一。这种革兰氏阴性菌的荚膜是一种主要的毒力因子,在防御和生物膜形成中起重要作用。噬菌体对一种细菌菌株及其荚膜类型具有特异性,在多糖解聚酶的辅助下可引起细菌细胞裂解。在本研究中,我们分离并鉴定了一种针对医院52145株K2荚膜血清型的噬菌体。该噬菌体显示出狭窄的宿主范围和稳定的裂解活性,即使暴露于不同温度或去污剂中也是如此。在体内研究了该噬菌体在鼻腔定植模型中的预防作用。系统发育分析表明,新分离的噬菌体B1属于科属。我们确定了新噬菌体荚膜解聚酶基因的位置,并对其进行了扩增、克隆、表达和纯化。通过在菌株上点样测试了重组B1dep解聚酶的功效,证实该提取物在降解细菌细胞上的保护性荚膜时降低了菌苔厚度。由于具有K2血清型的菌株具有流行病学重要性,B1噬菌体及其解聚酶有望用作可能的抗菌剂。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fc49/8003838/21674691ebdb/microorganisms-09-00650-g001.jpg

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