Identification and Characterization of Dpo42, a Novel Depolymerase Derived from the Escherichia coli Phage vB_EcoM_ECOO78.
Identification and Characterization of Dpo42, a Novel Depolymerase Derived from the Escherichia coli Phage vB_EcoM_ECOO78.
复制标题
Dpo42（一种源自大肠杆菌噬菌体 vB_EcoM_ECOO78 的新型解聚酶）的鉴定和表征
DOI：
10.3389/fmicb.2017.01460
复制
发表时间：
2017
影响因子：
5.2
通讯作者：
Yang J
中科院分区：
文献类型：
--
作者：
Guo Z;Huang J;Yan G;Lei L;Wang S;Yu L;Zhou L;Gao A;Feng X;Han W;Gu J;Yang J
生物膜的形成是致病菌最重要的毒力因子之一，可以保护细菌免受干燥、抗生素、细菌感染和宿主免疫反应的影响。然而，噬菌体衍生的解聚酶显示出生物膜活性，并显示出治疗由生物膜形成细菌引起的感染的巨大潜力。在这项研究中，大肠杆菌噬菌体vB_EcoM_ECOO78的分离和表征，我们观察到它的能力，裂解五个34个测试的E。大肠杆菌临床分离株。在10-5的感染复数和约74空斑形成单位（PFU）/感染的爆发大小下观察到最高噬菌体滴度。电子显微照片表明vB_EcoM_ECOO78属于肌尾病毒科。由vB_EcoM_ECOO78形成的裂解噬菌斑周围存在增加的晕圈，表明该噬菌体可能编码解聚酶。基于测序分析，发现vB_EcoM_ECOO78的全基因组大小为41，289 bp，GC含量为53.07%。此外，vB_EcoM_ECOO78有56个预测的开放阅读帧，其中51个（91.07%）被认为是功能性的。BLAST分析表明vB_EcoM_ECOO78（Dpo 42）的ORF 42与其他报道的噬菌体相关解聚酶具有低的同一性。Dpo 42在大肠杆菌中表达并纯化为可溶性蛋白。coli BL21。对E.大肠杆菌菌株和Dpo 42的膜活性通过进行斑点试验和使用96孔微量滴定板法进行测试。Dpo 42降解E.并表现出剂量依赖性的生物膜形成预防活性。基于这些结果，Dpo 42似乎是一种新的噬菌体来源的解聚酶，代表了一种新的潜在的预防大肠杆菌的策略。大肠杆菌生物膜形成。
Biofilm formation, one of the most important virulence factors of pathogenic bacteria, protects bacteria against desiccation, antibiotics, phages and host immune responses. However, phage-derived depolymerases show antibiofilm activity and demonstrate great potential to treat infections caused by biofilm-forming bacteria. In this study, the Escherichia coli phage vB_EcoM_ECOO78 was isolated and characterised, and we observed its ability to lyse five out of 34 tested E. coli clinical isolates. The highest phage titre was observed at a multiplicity of infection of 10-5 and a burst size of approximately 74 plaque forming units (PFU)/infection. Electron micrographs indicated that vB_EcoM_ECOO78 belongs to the family Myoviridae. The presence of increasing halos surrounding the lysis plaques formed by vB_EcoM_ECOO78 indicated that this phage may encode a depolymerase. Based on a sequencing analysis, the complete genome of vB_EcoM_ECOO78 was found to be 41,289 bp in size, with a GC content of 53.07%. Additionally, vB_EcoM_ECOO78 has 56 predicted open reading frames, 51 (91.07%) of which are assumed to be functional. A BLAST analysis indicated that ORF42 of vB_EcoM_ECOO78 (Dpo42) has low identity with other reported phage-associated depolymerases. Dpo42 was expressed and purified as a soluble protein using E. coli BL21. The biofilm formation ability of E. coli isolates and the antibiofilm activity of Dpo42 were tested by performing spot assays and using a 96-well micro-titre plate method. Dpo42 degraded the capsular polysaccharides surrounding E. coli and exhibited dose-dependent biofilm-formation prevention activity. Based on these results, Dpo42 appears to be a novel phage-derived depolymerase that represents a new potential strategy for preventing E. coli biofilm formation.