| 55 | 0 | 45 |
| 下载次数 | 被引频次 | 阅读次数 |
目的 构建一种新型α/β类肽嵌合体，并探讨其在体外环境下的抗菌效能与生化稳定性。方法 将β氨基酸引入，通过固相合成法合成了α与β氨基酸交替序列的α/β-PM，用高效液相色谱分析其纯度，质谱进行分子量验证。采用微量肉汤稀释法测定了该多肽对大肠埃希菌（E. coli）ATCC 25922、金黄色葡萄球菌（S. aureus）ATCC 29213的最小抑菌浓度（MIC）和最小杀菌浓度（MBC），用结晶紫染色法和稀释涂板法检测了多肽对成熟生物膜形成的抑制和清除效果。用时间杀菌曲线实验评价其杀菌速度和浓度依赖性特征。在E. coli模型上通过微量肉汤稀释法评估了其在蛋白酶环境下的稳定性。结果 α/β-PM的抗菌效果显著优于传统α抗菌肽LL-37和乳酸链球菌肽（Nisin），表现为杀菌活性，对S. aureus具有浓度依赖性杀菌作用。α/β-PM在MIC下即可显著抑制S. aureus生物膜形成（抑制率>90%），清除成熟生物膜能力也强于LL-37和Nisin。经胰蛋白酶、链霉蛋白酶E处理后，α/β-PM的MIC保持不变。结论 α/β-PM较传统抗菌肽具有更强的杀菌活性，对S. aureus呈浓度依赖性杀菌效果并可有效抑制和清除生物膜形成。此外，其具有优异的蛋白酶稳定性，经多种蛋白酶处理后仍维持抗菌效力，具备良好的临床转化潜力。
Abstract：Objective To constructed a novel α-peptide/β-peptoid peptidomimetics and evaluated its antibacterial efficacy and biochemical stability under in vitro conditions. Methods β-Amino acids were introduced， and α/β-PM with alternating α- and β-amino acid sequences was synthesized via solid-phase peptide synthesis. Its purity was analyzed by high-performance liquid chromatography (HPLC)， and its molecular weight was confirmed by mass spectrometry (MS). The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) against Escherichia coli (E. coli) ATCC 25922 and Staphylococcus aureus (S. aureus) ATCC 29213 were determined using the broth microdilution method. The inhibitory and eradication effects on mature biofilms were evaluated by crystal violet staining and dilution plating methods. Time-kill curve assays were used to assess its bactericidal rate and concentration-dependent characteristics. Its stability in the presence of proteases was evaluated using the broth microdilution method in an E. coli model. Results The antibacterial activity of α/β-PM was significantly superior to that of the conventional α-peptides LL-37 and Nisin. It exhibited potent bactericidal activity with concentration-dependent killing against S. aureus. At its MIC， α/β-PM inhibited S. aureus biofilm formation by >90% and showed stronger eradication of mature biofilms than LL-37 and nisin. After treatment with trypsin and proteinase E， the MIC of α/β-PM remained unchanged. Conclusion α/β-PM exhibited stronger bactericidal activity than conventional antimicrobial peptides， with concentration-dependent killing against S. aureus. It effectively inhibited and eradicated biofilm formation. Moreover， it displayed excellent protease stability， maintaining its antibacterial potency after treatment with multiple proteases， indicating strong potential for clinical translation.
[1] 李娇娇， 韦章澳， 张维波， 等. 钛表面载银的硫酸乙酰肝素/壳聚糖抗菌涂层的构建[J]. 安徽医科大学学报， 2022， 57(5): 754-8. doi:10.19405/j.cnki.issn1000-1492.2022.05.015.
[2] Ma X， Wang Q， Ren K， et al. A review of antimicrobial peptides: structure， mechanism of action， and molecular optimization strategies[J]. Fermentation， 2024， 10(11): 540. doi:10.3390/fermentation10110540.
[3] Hetta H F， Sirag N， Alsharif S M， et al. Antimicrobial peptides: the game-changer in the epic battle against multidrug-resistant bacteria[J]. Pharmaceuticals， 2024， 17(11): 1555. doi:10.3390/ph17111555.
[4] Sadeeq M， Li Y， Wang C， et al. Unlocking the power of antimicrobial peptides: advances in production， optimization， and therapeutics[J]. Front Cell Infect Microbiol， 2025， 15: 1528583. doi:10.3389/fcimb.2025.1528583.
[5] Zhu J， Gao L， Xue W， et al. Engineering β-amino acid antimicrobial peptides with enhanced stability and potential efficacy against gram-negative bacterial infections[J]. Eur J Med Chem， 2026， 310: 118802. doi:10.1016/j.ejmech.2026.118802.
[6] Voronko O E， Khotina V A， Kashirskikh D A， et al. Antimicrobial peptides of the cathelicidin family: focus on LL-37 and its modifications[J]. Int J Mol Sci， 2025， 26(16): 8103. doi:10.3390/ijms26168103.
[7] Jiang W， Zhou M， Chen K， et al. Biguanide-functionalized peptide mimics effectively combat drug-resistant ESKAPE pathogens and meningitis[J]. Nat Commun， 2025， 17(1): 336. doi:10.1038/s41467-025-67044-5.
[8] Zhang M， Ouyang J， Fu L， et al. Hydrophobicity determines the bacterial killing rate of α-helical antimicrobial peptides and influences the bacterial resistance development[J]. J Med Chem， 2022， 65(21): 14701-20. doi:10.1021/acs.jmedchem.2c01238.
[9] Bucataru C， Ciobanasu C. Antimicrobial peptides: opportunities and challenges in overcoming resistance[J]. Microbiol Res， 2024， 286: 127822. doi:10.1016/j.micres.2024.127822.
[10] Kocagoz T， Temur B Z， Unubol N， et al. Protease-resistant， broad-spectrum antimicrobial peptides with high antibacterial and antifungal activity[J]. Life， 2025， 15(2): 242. doi:10.3390/life15020242.
[11] Selvarajan V， Tram N D T， Xu J， et al. Stapled β-hairpin antimicrobial peptides with improved stability and activity against drug-resistant gram-negative bacteria[J]. J Med Chem， 2023， 66(13): 8498-509. doi:10.1021/acs.jmedchem.3c00140.
[12] Liu H Y， Prentice E L， Webber M A. Mechanisms of antimicrobial resistance in biofilms[J]. npj Antimicrob Resist， 2024， 2: 27. doi:10.1038/s44259-024-00046-3.
[13] Kalia V C， Patel S K S， Lee J K. Bacterial biofilm inhibitors: an overview[J]. Ecotoxicol Environ Saf， 2023， 264: 115389. doi:10.1016/j.ecoenv.2023.115389.
[14] Li X， Lin S， Wang Y， et al. Application of biofilm dispersion-based nanoparticles in cutting off reinfection[J]. Appl Microbiol Biotechnol， 2024， 108(1): 386. doi:10.1007/s00253-024-13120-7.
[15] Zheng S， Tu Y， Li B， et al. Antimicrobial peptide biological activity， delivery systems and clinical translation status and challenges[J]. J Transl Med， 2025， 23(1): 292. doi:10.1186/s12967-025-06321-9.
基本信息:
中图分类号:R965
引用信息:
[1]邢健远,陶鑫,张志浩,等.耐酶解α/β类肽嵌合体的体外抗菌活性研究[J].安徽医科大学学报().
基金信息:
安徽省自然科学基金（编号：2208085QE136）
2026-08-27
2026-08-27
2026-08-27