Centre of Biological Engineering, University of Minho, Braga, Portugal.
LABBELS -Associate Laboratory, Braga, Guimarães, Portugal.
Appl Microbiol Biotechnol. 2023 Jun;107(11):3621-3636. doi: 10.1007/s00253-023-12547-8. Epub 2023 May 3.
Citrobacter koseri is an emerging Gram-negative bacterial pathogen, which causes urinary tract infections. We isolated and characterized a novel S16-like myovirus CKP1 (vB_CkoM_CkP1), infecting C. koseri. CkP1 has a host range covering the whole C. koseri species, i.e., all strains that were tested, but does not infect other species. Its linear 168,463-bp genome contains 291 coding sequences, sharing sequence similarity with the Salmonella phage S16. Based on surface plasmon resonance and recombinant green florescence protein fusions, the tail fiber (gp267) was shown to decorate C. koseri cells, binding with a nanomolar affinity, without the need of accessory proteins. Both phage and the tail fiber specifically bind to bacterial cells by the lipopolysaccharide polymer. We further demonstrate that CkP1 is highly stable towards different environmental conditions of pH and temperatures and is able to control C. koseri cells in urine samples. Altogether, CkP1 features optimal in vitro characteristics to be used both as a control and detection agent towards drug-resistant C. koseri infections. KEY POINTS: • CkP1 infects all C. koseri strains tested • CkP1 recognizes C. koseri lipopolysaccharide through its long tail fiber • Both phage CkP1 and its tail fiber can be used to treat or detect C. koseri pathogens.
柠檬酸杆菌是一种新兴的革兰氏阴性细菌病原体,可引起尿路感染。我们分离并鉴定了一种新型的 S16 样肌病毒 CKP1(vB_CkoM_CkP1),可感染柠檬酸杆菌。CkP1 的宿主范围涵盖整个柠檬酸杆菌种,即所有测试的菌株,但不感染其他种。其线性 168463bp 基因组包含 291 个编码序列,与沙门氏菌噬菌体 S16 具有序列相似性。基于表面等离子体共振和重组绿色荧光蛋白融合,发现尾丝(gp267)装饰柠檬酸杆菌细胞,以纳摩尔亲和力结合,而不需要辅助蛋白。噬菌体和尾丝都通过脂多糖聚合物特异性地结合到细菌细胞上。我们进一步证明,CkP1 对不同的 pH 值和温度环境条件具有高度稳定性,并且能够控制尿液样本中的柠檬酸杆菌细胞。总的来说,CkP1 具有最佳的体外特性,可作为控制和检测耐药性柠檬酸杆菌感染的试剂。关键点:• CkP1 可感染所有测试的柠檬酸杆菌菌株• CkP1 通过其长尾丝识别柠檬酸杆菌脂多糖• 噬菌体 CkP1 及其尾丝均可用于治疗或检测柠檬酸杆菌病原体。