Oyama Linda B, Crochet Jean-Adrien, Edwards Joan E, Girdwood Susan E, Cookson Alan R, Fernandez-Fuentes Narcis, Hilpert Kai, Golyshin Peter N, Golyshina Olga V, Privé Florence, Hess Matthias, Mantovani Hilario C, Creevey Christopher J, Huws Sharon A
Institute of Biological Environmental and Rural Sciences, Aberystwyth UniversityAberystwyth, United Kingdom.
Institute of Infection and Immunity, St George's University of LondonLondon, United Kingdom.
Front Chem. 2017 Jul 12;5:51. doi: 10.3389/fchem.2017.00051. eCollection 2017.
Antimicrobial peptides (AMPs) are gaining popularity as alternatives for treatment of bacterial infections and recent advances in technologies provide new platforms for AMP discovery. We sought to determine the antibacterial activity of a novel antimicrobial peptide, buwchitin, against . Buwchitin was identified from a rumen bacterial metagenome library, cloned, expressed and purified. The antimicrobial activity of the recombinant peptide was assessed using a broth microdilution susceptibility assay to determine the peptide's killing kinetics against selected bacterial strains. The killing mechanism of buwchitin was investigated further by monitoring its ability to cause membrane depolarization (diSC(5) method) and morphological changes in cells. Transmission electron micrographs of buwchitin treated cells showed intact outer membranes with blebbing, but no major damaging effects and cell morphology changes. Buwchitin had negligible cytotoxicity against defibrinated sheep erythrocytes. Although no significant membrane leakage and depolarization was observed, buwchitin at minimum inhibitory concentration (MIC) was bacteriostatic against cells and inhibited growth by 70% when compared to untreated cells. These findings suggest that buwchitin, a rumen derived peptide, has potential for antimicrobial activity against .
抗菌肽（AMPs）作为治疗细菌感染的替代物正日益受到关注，并且技术的最新进展为抗菌肽的发现提供了新平台。我们试图确定一种新型抗菌肽——布氏几丁质对……的抗菌活性。布氏几丁质是从瘤胃细菌宏基因组文库中鉴定出来的，经过克隆、表达和纯化。使用肉汤微量稀释药敏试验评估重组肽的抗菌活性，以确定该肽对选定细菌菌株的杀伤动力学。通过监测其引起膜去极化的能力（diSC(5)法）和……细胞的形态变化，进一步研究布氏几丁质的杀伤机制。布氏几丁质处理后的……细胞的透射电子显微镜图像显示，外膜完整但有泡状突起，但没有主要的损伤效应和细胞形态变化。布氏几丁质对去纤维蛋白羊红细胞的细胞毒性可忽略不计。虽然未观察到明显的膜泄漏和去极化现象，但最低抑菌浓度（MIC）的布氏几丁质对……细胞具有抑菌作用，与未处理细胞相比，生长抑制率达70%。这些发现表明，布氏几丁质这种源自瘤胃的肽具有对……的抗菌活性潜力。
