中文摘要：受弯曲杆菌污染的家禽是全世界食源性肠胃炎的主要病因，需要新型抗菌药物。我们先前提出了融合细胞内溶素与噬菌体受体结合蛋白（RBP）形成Innolysin的概念，并证明Innolysins对大肠杆菌具有杀菌活性。本文将Innolysin应用到空肠弯曲菌。由于迄今未发现空肠弯曲菌噬菌体RBP，我们首先发现了源于空肠弯曲菌CAMSA2147类CJIK1原噬菌体的H-纤维，其功能相当于新的RBP。通过融合H-纤维与T5噬菌体细胞内溶素，构建了针对空肠弯曲菌的Innolysins（Innolysins Cj）。20℃体外接触45分钟，Innolysin Cj1对不同的空肠弯曲菌株具有抗菌活性，CAMSA2147细胞计数减少到1.30 +/- 0.21log。通过筛选由不同细胞内溶素组成的抑制生长的Innolysins Cj库，确定细胞内溶素Innolysins Cj5作为另一种有希望的抗菌候选菌。将Innolysin Cj1或InnolysinCj5应用于5℃冷藏的空肠弯曲菌CAMSA2147污染的鸡皮上，细胞数量分别减少了1.63+/-0.46log和1.18+/-0.10log，证实了Innolysin Cj可以原位杀死空肠弯曲菌。Innolysins Cj的受体仍有待鉴定，然而，与结合荚膜多糖或鞭毛的裂解噬菌体相比，RBP成分(H-纤维)可识别一种新的受体。鉴定其他未开发的弯曲杆菌噬菌体RBPs，可能进一步增加针对不同受体的新型Innolysins Cj，并发挥对弯曲杆菌的抗菌作用。
外文摘要：Campylobacter contaminated poultry remains the major cause of foodborne gastroenteritis worldwide, calling for novel antibacterials. We previously developed the concept of Innolysin composed of an endolysin fused to a phage receptor binding protein (RBP) and provided the proof-of-concept that Innolysins exert bactericidal activity against Escherichia coli. Here, we have expanded the Innolysin concept to target Campylobacter jejuni. As no C. jejuni phage RBP had been identified so far, we first showed that the H-fiber originating from a CJIE1-like prophage of C. jejuni CAMSA2147 functions as a novel RBP. By fusing this H-fiber to phage T5 endolysin, we constructed Innolysins targeting C. jejuni (Innolysins Cj). Innolysin Cj1 exerts antibacterial activity against diverse C. jejuni strains after in vitro exposure for 45 min at 20 degrees C, reaching up to 1.30 +/- 0.21 log reduction in CAMSA2147 cell counts. Screening of a library of Innolysins Cj composed of distinct endolysins for growth inhibition, allowed us to select Innolysin Cj5 as an additional promising antibacterial candidate. Application of either Innolysin Cj1 or Innolysin Cj5 on chicken skin refrigerated to 5 degrees C and contaminated with C. jejuni CAMSA2147 led to 1.63 +/- 0.46 and 1.18 +/- 0.10 log reduction of cells, respectively, confirming that Innolysins Cj can kill C. jejuni in situ. The receptor of Innolysins Cj remains to be identified, however, the RBP component (H-fiber) recognizes a novel receptor compared to lytic phages binding to capsular polysaccharide or flagella. Identification of other unexplored Campylobacter phage RBPs may further increase the repertoire of new Innolysins Cj targeting distinct receptors and working as antibacterials against Campylobacter.
外文关键词：Campylobacter；prophage binding；endolysin；Innolysin；antibacterials；food safety
作者：Zampara, A；Sorensen, MCH；Gencay, YE；Grimon, D；Kristiansen, SH；Jorgensen, LS；Kristensen, JR；Briers, Y；Elsser-Gravesen, A；Brondsted, L
作者单位：Univ Copenhagen；Univ Ghent；ISI Food Protect ApS
期刊名称：FRONTIERS IN MICROBIOLOGY
期刊影响因子：4.236
出版年份：2021
出版刊次：21
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