The emergence of antibiotic-resistant microbes has stimulated research worldwide seeking new biologically active molecules. In this respect, synthetic antimicrobial peptides (SAMPs) have been suggested to overcome this problem. Although there are some online servers that provide putative SAMPs from protein sequences, the choice of the best peptide sequences for further analysis is still difficult. Therefore, the goal of this paper is not to launch a new tool but to provide a friendly workflow to characterize and predict potential SAMPs by employing existing tools. Using this proposed workflow, two peptides (PepGAT and PepKAA) were obtained and extensively characterized. These peptides damaged microbial membranes and cell walls, and induced overproduction of reactive oxygen species (ROS). Both peptides were found to assume random coil secondary structure in aqueous solution, organic solvent, and upon binding to negatively charged lipid systems. Peptides were also able to degrade formed biofilms but not to prevent biofilm formation. PepGAT was not resistant to proteolysis, whereas PepKAA was resistant to pepsin but not to pancreatin. Furthermore, both presented no hemolytic activity against red blood cells, even at a 10-fold higher concentration than the antimicrobial concentration. The pipeline proposed here is an easy way to design new SAMPs for application as alternatives to develop new drugs against human pathogenic microorganisms.

抗生素抗性微生物的出现刺激了全世界寻求新的生物活性分子的研究。在这方面，已经提出了合成的抗微生物肽（SAMP）来克服这个问题。尽管有一些在线服务器可以从蛋白质序列中提供推定的SAMP，但仍然难以选择最佳的肽序列进行进一步分析。因此，本文的目标不是启动新工具，而是提供一个友好的工作流程，以通过使用现有工具来表征和预测潜在的SAMP。使用提出的工作流程，获得了两种肽（PepGAT和PepKAA）并进行了广泛表征。这些肽破坏了微生物膜和细胞壁，并诱导了活性氧（ROS）的过量产生。发现这两种肽在水溶液，有机溶剂中以及与带负电的脂质系统结合后均呈现无规卷曲二级结构。肽也能够降解形成的生物膜，但不能阻止生物膜的形成。PepGAT对蛋白水解没有抵抗力，而PepKAA对胃蛋白酶却对胰酶没有抵抗力。此外，即使在浓度比抗微生物剂浓度高10倍的情况下，它们都没有针对红细胞的溶血活性。此处提出的管道是设计新SAMP的简便方法，可作为开发针对人类病原微生物的新药物的替代品。PepGAT对蛋白水解没有抵抗力，而PepKAA对胃蛋白酶却对胰酶没有抵抗力。此外，即使在浓度比抗微生物剂浓度高10倍的情况下，它们都没有针对红细胞的溶血活性。此处提出的管道是设计新SAMP的简便方法，可作为开发针对人类病原微生物的新药物的替代品。PepGAT对蛋白水解没有抵抗力，而PepKAA对胃蛋白酶却对胰酶没有抵抗力。此外，即使在浓度比抗微生物剂浓度高10倍的情况下，它们都没有针对红细胞的溶血活性。此处提出的管道是设计新SAMP的简便方法，可作为开发针对人类病原微生物的新药物的替代品。