中文摘要：噬菌体(内)细胞裂解酶是目前最有希望摆脱抗生素耐药性危机的方法之一。无论作为单一疗法还是作为普通抗生素化疗的补充，细胞裂解酶均已进入后期临床阶段，将获得监管机构的批准。细胞裂解酶不能从外部感染革兰氏阴性细菌的问题，也已通过各种方式克服：通过工程方法或研究某些野生型细胞裂解酶与细菌表面相互作用的自然机制。某些细胞裂解酶的这种内在能力与类抗菌肽(AMP)区域有关，而这些区域本身就是新型抗菌素的重要来源。然而，目前许多寻找新型细胞裂解酶类抗生素候选药物都依赖于实验筛选。本研究对侵染革兰氏阴性假单胞菌属的噬菌体一组细胞裂解酶的C末端进行了生物信息学分析。通过计算理化性质，利用预测k-最近邻(kNN)模型预估该区域成为AMP的概率，从原始数据库中获得了一组假定的膜相互作用细胞裂解酶。对其中两个候选药物（命名为Pae87和Ppl65）在溶壁、溶菌和杀菌活性方面进行了前瞻性测试。发现二者都对假单胞菌和其他革氏阴性细菌病原体具有活性，说明该方法对预测类AMP区域有用,可用于开发噬菌体细胞裂解酶,设计和开发出抗菌素或抗菌部分。
外文摘要：Phage (endo)lysins are nowadays one of the most promising ways out of the current antibiotic resistance crisis. Either as sole therapeutics or as a complement to common antibiotic chemotherapy, lysins are already entering late clinical phases to get regulatory agencies' authorization. Even the old paradigm of the inability of lysins to attack Gram-negative bacteria from without has already been overcome in a variety of ways: either by engineering approaches or investigating the natural mechanisms by which some wild-type lysins are able to interact with the bacterial surface. Such inherent ability of some lysins has been linked to antimicrobial peptide (AMP)-like regions, which are, on their own, a significant source for novel antimicrobials. Currently, though, many of the efforts for searching novel lysin-based antimicrobial candidates rely on experimental screenings. In this work, we have bioinformatically analyzed the C-terminal end of a collection of lysins from phages infecting the Gram-negative genus Pseudomonas. Through the computation of physicochemical properties, the probability of such regions to be an AMP was estimated by means of a predictive k-nearest neighbors (kNN) model. This way, a subset of putatively membrane-interacting lysins was obtained from the original database. Two of such candidates (named Pae87 and Ppl65) were prospectively tested in terms of muralytic, bacteriolytic, and bactericidal activity. Both of them were found to possess an activity against Pseudomonas aeruginosa and other Gram-negative bacterial pathogens, implying that the prediction of AMP-like regions could be a useful approach toward the mining of phage lysins to design and develop antimicrobials or antimicrobial parts for further engineering.
外文关键词：lysins；Gram-negative bacteria；bioinformatic analysis；enzybiotics； antimicrobials；enzyme mining；Pseudomonas aeruginosa
作者：Vazquez, R；Blanco-Ganan, S；Ruiz, S；Garcia, P
作者单位：CSIC；Ctr Invest Biomed Red Enfermedades Resp CIBERES
期刊名称：FRONTIERS IN MICROBIOLOGY
期刊影响因子：4.236
出版年份：2021
出版刊次：12
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