Application of HPCCC Combined with Polymeric Resins and HPLC for the Separation of Cyclic Lipopeptides Muscotoxins A-C and Their Antimicrobial Activity
Application of HPCCC Combined with Polymeric Resins and HPLC for the Separation of Cyclic Lipopeptides Muscotoxins A-C and Their Antimicrobial Activity
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DOI：
10.3390/molecules23102653
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发表时间：
2018-10-01
期刊：
影响因子：
4.6
通讯作者：
Hrouzek, Pavel
中科院分区：
文献类型：
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作者：
Cheel, Jose;Hajek, Jan;Hrouzek, Pavel
粘菌毒素是蓝藻环状脂肽，在生物医学和生物技术中具有潜在的应用。本研究通过聚合树脂处理富集麝香CCALA125菌株提取物，并进行HPCCC，得到三种环状脂肽(1-3)，通过半制备型HPLC进一步纯化，得到1、2和3，纯度分别为86%、92%和90%。通过与之前报道的 ESI-HRMS/MS 数据比较，2-3 的化学特性分别确定为粘菌毒素 A 和 B，而利用 NMR 和 ESI-HRMS/MS 数据确定 1 为新型粘菌毒素变体（粘菌毒素 C）。由于产量高（50 mg），化合物 2 被广泛筛选其抗菌潜力，对链格孢、黄瓜单孢菌和烟曲霉表现出强大的抗真菌活性，最低抑菌浓度 (MIC) 值为 0.58、2.34 和 2.34 mu g/mL；对枯草芽孢杆菌的抗菌活性较弱，MIC值为37.5μg/mL。由于产量低，化合物 1 和 3 仅针对植物病原真菌核盘菌进行了测试，显示出中等的抗真菌活性。所开发的色谱方法被证明是获得具有有效抗真菌特性的粘菌毒素的有效工具。
Muscotoxins are cyanobacterial cyclic lipopeptides with potential applications in biomedicine and biotechnology. In this study, Desmonostoc muscorum CCALA125 strain extracts were enriched by polymeric resin treatment, and subjected to HPCCC affording three cyclic lipopeptides (1-3), which were further repurified by semi-preparative HPLC, affording 1, 2, and 3, with a purity of 86%, 92%, and 90%, respectively. The chemical identities of 2-3 were determined as muscotoxins A and B, respectively, by comparison with previously reported ESI-HRMS/MS data, whereas 1 was determined as a novel muscotoxin variant (muscotoxin C) using NMR and ESI-HRMS/MS data. Owing to the high yield (50 mg), compound 2 was broadly screened for its antimicrobial potential exhibiting a strong antifungal activity against Alternaria alternata, Monographella cucumerina, and Aspergillus fumigatus, with minimum inhibitory concentration (MIC) values of 0.58, 2.34, and 2.34 mu g/mL; respectively, and weak antibacterial activity against Bacillus subtilis with a MIC value of 37.5 mu g/mL. Compounds 1 and 3 were tested only against the plant pathogenic fungus Sclerotinia sclerotiorum due to their low yield, displaying a moderate antifungal activity. The developed chromatographic method proved to be an efficient tool for obtaining muscotoxins with potent antifungal properties.