抗菌肽Loong-3的分子设计及其抑菌活性影响因素分析

doi:10.16431/j.cnki.1671-7236.2025.10.040

摘要/Abstract

摘要： 【目的】通过分子设计及抑菌性能评价，获得具有广谱抗菌活性的抗菌肽，为临床生产提供新的抗生素替代品。【方法】借助在线生物信息学工具设计1条多肽Loong-3，采用琼脂糖扩散法测定其抑菌活性，通过倍比稀释法测定其最小抑菌浓度，采用菌落培养法测定其最小杀菌浓度，通过倍比稀释法测定其溶血率，并分析多种理化因子(热、紫外线、反复冻融、盐离子、酸碱、血清、有机试剂、酶)对其抑菌活性的影响。【结果】设计的多肽Loong-3为新的阳离子型疏水性耐热螺旋多肽，抗菌谱较广，其对革兰阴性菌大肠杆菌、沙门菌和绿脓杆菌的最小抑菌浓度和最小杀菌浓度分别为0.976～500和1.953～31.25 μg/mL，对革兰阳性菌金黄色葡萄球菌的最小抑菌浓度和最小杀菌浓度分别为31.25和62.5 μg/mL，对真菌白色念珠菌的最小抑菌浓度和最小杀菌浓度分别为125和500 μg/mL。多肽Loong-3有一定的溶血性，浓度从0.976增至250 μg/mL时，溶血率也从5.78%升至76.68%。除了胰酶外，在多种理化因子(热、紫外线、反复冻融、盐离子、酸碱、血清、有机试剂、酶)作用下，多肽Loong-3的最小抑菌浓度虽有波动，但仍具有抑菌活性。【结论】设计的多肽Loong-3为新的阳离子型广谱抗菌肽，有一定的溶血性，热、紫外、反复冻融、盐离子、酸碱、血清、有机试剂、酶对多肽Loong-3的抑菌活性没有明显影响。

关键词: 抗菌肽; 分子设计; 抑菌活性; 生物信息学; 抗生素替代品

Abstract: 【Objective】 This study aimed to obtain antimicrobial peptides with broad-spectrum antimicrobial activity through molecular design and antibacterial performance evaluation,and provide a new antibiotic alternatives for clinical production.【Method】 A polypeptide named Loong-3 was designed using online bioinformatics tools.The antimicrobial activity,minimal inhibitory concentration (MIC),minimum bactericidal concentration (MBC) and hemolysis rate of polypeptide Loong-3 were determined by agarose diffusion method,double dilution method,colony culture method and double dilution method,respectvely.And the effects of various physicochemical factors (heat,UV,repeated freeze-thaw,salt ions,acid-base,serum,organic reagents,enzymes) on the antimicrobial activity of polypeptide Loong-3 were analyzed.【Result】 The results showed that the polypeptide Loong-3 was a new cationic hydrophobic heat-resistant spiral peptide with a broad antimicrobial spectrum.Its MIC and MBC against Gram-negative bacteria Escherichia coli,Salmonella and Pseudomonas aeruginosa were 0.976-500 and 1.953-31.25 μg/mL,respectively,the MIC and MBC against Gram-positive bacteria Staphylococcus aureus were 31.25 and 62.5 μg/mL,respectively,and the MIC and MBC against fungi Candida albicans were 125 and 500 μg/mL,respectively.Peptide Loong-3 had a certain degree of hemolysis,and when the concentration of polypeptide Loong-3 increased from 0.976 to 250 μg/mL,the hemolysis rate also increased from 5.78% to 76.68%.Except for trypsin,the polypeptide Loong-3 still had antimicrobial activity under various physical and chemical factors (heat,UV,repeated freeze-thaw,salt ions,acid-base,serum,organic reagents,enzymes) though the MIC was fluctuated.【Conclusion】 The designed polypeptide Loong-3 was a new cationic broad-spectrum antimicrobial peptide with certain hemolytic properties.Heat,UV,repeated freeze-thaw cycles,salt ions,acid-base,serum,organic reagents,and enzymes had no significant effect on the antimicrobial activity of polypeptide Loong-3.

Key words: antimicrobial polypeptide; molecular design; antibacterial activity; bioinformatics; antibiotic alternatives

中图分类号:

S859.79⁺7

杭柏林, 李杰, 阿卜杜瓦日斯·艾海提, 程洛妤, 楚彪, 董明欣, 任涛, 盛玥, 宋如意, 王俊文, 何宏轩, 刘保国. 抗菌肽Loong-3的分子设计及其抑菌活性影响因素分析[J]. 中国畜牧兽医, 2025, 52(10): 4952-4963.

HANG Bolin, LI Jie, ABDUWARIS Aihaiti, CHENG Luoyu, CHU Biao, DONG Mingxin, REN Tao, SHENG Yue, SONG Ruyi, WANG Junwen, HE Hongxuan, LIU Baoguo. Molecular Design of Antimicrobial Peptide Loong-3 and Analysis of Factors Influencing Its Antimicrobial Activity[J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(10): 4952-4963.

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