Tasdurmazli Semra, Dokuz Senanur, Erdogdu Berna, Var Isil, Chen John Yu-Shen, Ozbek Tulin
Department of Molecular Biology and Genetics, Faculty of Arts and Sciences, Yildiz Technical University, Istanbul, Turkey.
Department of Food Engineering, Faculty of Agricultural, Cukurova University, Sarıcam-Adana, Turkey.
Biotechnol J. 2023 Oct;18(10):e2300145. doi: 10.1002/biot.202300145. Epub 2023 Jun 23.
Bacteriophages, which selectively infect bacteria, and phage-derived structures are considered promising agents for the diagnosis and treatment of bacterial infections due to the increasing antibiotic resistance. The binding of phages to their specific receptors on host bacteria is highly specific and irreversible, and therefore, the characterization of receptor-binding proteins(RBPs), which are key determinants of phage specificity, is crucial for the development of new diagnostic and therapeutic products. This study highlights the biotechnological potential of Gp144, an RBP located in the tail baseplate of bacteriophage K and responsible for adsorption of phageK to S. aureus. Once it was established that recombinant Gp144 (rGp144)is biocompatible and does not exhibit lytic effects on bacteria, its interaction with the host, the binding efficiency and performance were assessed in vitro using microscopic and serological methods. Results showed that rGp144 has a capture efficiency (CE) of over 87% and the best CE score is %96 which captured 9 CFU mL out of 10 CFU mL bacteria, indicating that very low number of bacteria could be detected. Additionally, it was shown for the first time in the literature that rGp144 binds to both S. aureus and methicillin-resistant S. aureus (MRSA) cells in vitro, while its affinity to different Gram-positive bacteria (E. faecalis and B. cereus) was not observed. The findings suggest that rGp144 can be effectively used for the diagnosis of S. aureus and MRSA, and that the use of RBPs in host-phage interaction can be a novel and effective strategy for imaging and diagnosing the site of infection.
噬菌体可选择性感染细菌，鉴于抗生素耐药性不断增加，噬菌体及其衍生结构被视为诊断和治疗细菌感染的有前景的制剂。噬菌体与宿主细菌上其特定受体的结合具有高度特异性且不可逆，因此，作为噬菌体特异性关键决定因素的受体结合蛋白（RBPs）的特性表征，对于开发新的诊断和治疗产品至关重要。本研究突出了Gp144的生物技术潜力，Gp144是位于噬菌体K尾基板上的一种RBP，负责噬菌体K吸附到金黄色葡萄球菌上。一旦确定重组Gp144（rGp144）具有生物相容性且对细菌不表现出裂解作用，便使用显微镜和血清学方法在体外评估其与宿主的相互作用、结合效率和性能。结果表明，rGp144的捕获效率（CE）超过87%，最佳CE分数为96%，即从每毫升10个菌落形成单位（CFU）的细菌中捕获了9个CFU，这表明可以检测到数量非常少的细菌。此外，文献中首次表明rGp144在体外可与金黄色葡萄球菌和耐甲氧西林金黄色葡萄球菌（MRSA）细胞结合，而未观察到其对不同革兰氏阳性菌（粪肠球菌和蜡样芽孢杆菌）的亲和力。这些发现表明，rGp144可有效地用于金黄色葡萄球菌和MRSA的诊断，并且在宿主 - 噬菌体相互作用中使用RBPs可能是一种用于成像和诊断感染部位的新颖且有效的策略。
