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铜绿假单胞菌噬菌体 vB_PaeP_Lx18 的特性分析与基因组学研究及其溶菌酶的抑菌活性

Characterization and genome analysis of Pseudomonas aeruginosa phage vB_PaeP_Lx18 and the antibacterial activity of its lysozyme.

机构信息

College of Veterinary Medicine, Qingdao Agricultural University, Shandong, 266109, China.

出版信息

Arch Virol. 2022 Sep;167(9):1805-1817. doi: 10.1007/s00705-022-05472-0. Epub 2022 Jun 18.

Abstract

A lytic Pseudomonas aeruginosa phage, vB_PaeP_Lx18 (Lx18), was isolated from the sewage of a dairy farm. Biological characterization revealed that Lx18 was stable from 40 °C to 60 °C and over a wide range of pH values from 4 to 10. It was able to lyse 63.6% (21/33) of the P. aeruginosa strains tested and was able to reduce and disperse biofilms, with a biofilm reduction rate of 76.8%. Whole-genome sequencing showed that Lx18 is a dsDNA virus with a genome of 42,735 bp and G+C content of 62.16%. The genome contains 54 open reading frames (ORFs), 28 of which have known functions, including DNA replication and modification, transcriptional regulation, structural and packaging proteins, and host cell lysis. No virulence or tRNA genes were identified. Phylogenetic analysis showed that phage Lx18 belongs to the genus Phikmvvirus. The lysozyme of Lx18, Lys18, was cloned and expressed. The combined action of Lys18 and ethylenediaminetetraacetic acid (EDTA) had antibacterial activity against Pseudomonas aeruginosa. The study of phage Lx18 and its lysozyme will provide basic information for further research on the treatment of Pseudomonas aeruginosa infections.

摘要

一株裂解性铜绿假单胞菌噬菌体 vB_PaeP_Lx18(Lx18)从一家奶牛场的污水中分离得到。生物学特性表明,Lx18 在 40°C 至 60°C 以及 pH 值 4 至 10 的广泛范围内均稳定。它能够裂解 63.6%(21/33)测试的铜绿假单胞菌菌株,并能够减少和分散生物膜,生物膜减少率为 76.8%。全基因组测序表明,Lx18 是一种双链 DNA 病毒,基因组大小为 42735bp,G+C 含量为 62.16%。基因组包含 54 个开放阅读框(ORFs),其中 28 个具有已知功能,包括 DNA 复制和修饰、转录调控、结构和包装蛋白以及宿主细胞裂解。未鉴定出毒力或 tRNA 基因。系统发育分析表明,噬菌体 Lx18 属于 Phikmvvirus 属。克隆并表达了 Lx18 的溶菌酶 Lys18。Lys18 和乙二胺四乙酸(EDTA)的联合作用对铜绿假单胞菌具有抗菌活性。对噬菌体 Lx18 及其溶菌酶的研究将为进一步研究铜绿假单胞菌感染的治疗提供基础信息。

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