Abstract：
BACKGROUND: The consistently increasing reports of bacterial resistance and the reemergence of bacterial epidemics have inspired the health and scientific community to discover new molecules with antibacterial potential continuously. Frog-skin secretions constitute bioactive compounds essential for finding new biopharmaceuticals. The exact antibacterial characterization of dermaseptin related peptides derived from Agalychnis annae, is limited. The resemblance in their conserved and functionally linked genomes indicates an unprecedented opportunity to obtain novel bioactive compounds.
OBJECTIVE: In this study, we derived a novel peptide sequence and determined its antibacterial potentials.
METHODS: Consensus sequence strategy was used to design the novel and active antibacterial peptide named \'AGAAN\' from skin secretions of Agalychnis annae. The in-vitro activities of the novel peptide against some bacterial strains were investigated. Time kill studies, DNA retardation, cytotoxicity, betagalactosidase, and molecular computational studies were conducted.
RESULTS: AGAAN inhibited P. aeruginosa, E. faecalis, and S. typhimurium at 20 μM concentration. E. coli and S. aureus were inhibited at 25 μM, and lastly, B. subtilis at 50 μM. Kinetics of inactivation against exponential and stationary growing bacteria was found to be rapid within 1-5 hours of peptide exposure, depending on time and concentration. The peptide displayed weak hemolytic activity between 0.01%-7.31% at the antibacterial concentrations. AGAAN efficiently induced bacterial membrane damage with subsequent cell lysis. The peptide\'s DNA binding shows that it also targets intracellular DNA by retarding its movement. Our in-silico molecular docking analysis displayed a strong affinity to the bacterial cytoplasmic membrane.
CONCLUSIONS: AGAAN exhibits potential antibacterial properties that could be used to combat bacterial resistance.
OBJECTIVE: In this study, we derived a novel peptide sequence and determined its antibacterial potentials.
METHODS: Consensus sequence strategy was used to design the novel and active antibacterial peptide named \'AGAAN\' from skin secretions of Agalychnis annae. The in-vitro activities of the novel peptide against some bacterial strains were investigated. Time kill studies, DNA retardation, cytotoxicity, betagalactosidase, and molecular computational studies were conducted.
RESULTS: AGAAN inhibited P. aeruginosa, E. faecalis, and S. typhimurium at 20 μM concentration. E. coli and S. aureus were inhibited at 25 μM, and lastly, B. subtilis at 50 μM. Kinetics of inactivation against exponential and stationary growing bacteria was found to be rapid within 1-5 hours of peptide exposure, depending on time and concentration. The peptide displayed weak hemolytic activity between 0.01%-7.31% at the antibacterial concentrations. AGAAN efficiently induced bacterial membrane damage with subsequent cell lysis. The peptide\'s DNA binding shows that it also targets intracellular DNA by retarding its movement. Our in-silico molecular docking analysis displayed a strong affinity to the bacterial cytoplasmic membrane.
CONCLUSIONS: AGAAN exhibits potential antibacterial properties that could be used to combat bacterial resistance.
摘要：
背景：不断增加的细菌耐药性和细菌流行病的再次出现的报道激发了健康和科学界不断发现具有抗菌潜力的新分子。青蛙-皮肤分泌物构成了发现新的生物药物所必需的生物活性化合物。来自Agalychnisannae的dermaseptin相关肽的精确抗菌特性，是有限的。它们的保守和功能连接的基因组的相似性表明获得新的生物活性化合物的前所未有的机会。
目的：在本研究中，我们得到了一个新的肽序列，并确定了其抗菌潜力。
方法：采用共有序列策略，从Agalychnisannae的皮肤分泌物中设计出新型活性抗菌肽，命名为“AGAAN”。研究了新型肽对某些细菌菌株的体外活性。消磨时间的研究，DNA阻滞,细胞毒性，β-半乳糖苷酶，并进行了分子计算研究。
结果：AGAAN抑制铜绿假单胞菌，E.粪便，和20μM浓度的鼠伤寒沙门氏菌。大肠杆菌和金黄色葡萄球菌在25μM时被抑制，最后，50μM的枯草芽孢杆菌。发现针对指数和固定生长的细菌的失活动力学在肽暴露的1-5小时内迅速。取决于时间和浓度。该肽在抗菌浓度下表现出0.01%-7.31%之间的弱溶血活性。AGAAN有效诱导细菌膜损伤，随后细胞裂解。肽的DNA结合表明它还通过延缓其运动而靶向细胞内DNA。我们的计算机分子对接分析显示出对细菌细胞质膜的强亲和力。
结论：AGAAN具有潜在的抗菌特性，可用于对抗细菌耐药性。
目的：在本研究中，我们得到了一个新的肽序列，并确定了其抗菌潜力。
方法：采用共有序列策略，从Agalychnisannae的皮肤分泌物中设计出新型活性抗菌肽，命名为“AGAAN”。研究了新型肽对某些细菌菌株的体外活性。消磨时间的研究，DNA阻滞,细胞毒性，β-半乳糖苷酶，并进行了分子计算研究。
结果：AGAAN抑制铜绿假单胞菌，E.粪便，和20μM浓度的鼠伤寒沙门氏菌。大肠杆菌和金黄色葡萄球菌在25μM时被抑制，最后，50μM的枯草芽孢杆菌。发现针对指数和固定生长的细菌的失活动力学在肽暴露的1-5小时内迅速。取决于时间和浓度。该肽在抗菌浓度下表现出0.01%-7.31%之间的弱溶血活性。AGAAN有效诱导细菌膜损伤，随后细胞裂解。肽的DNA结合表明它还通过延缓其运动而靶向细胞内DNA。我们的计算机分子对接分析显示出对细菌细胞质膜的强亲和力。
结论：AGAAN具有潜在的抗菌特性，可用于对抗细菌耐药性。
