单点氨基酸替换对杂合肽KL-21生物活性的影响

doi:10.16431/j.cnki.1671-7236.2025.05.042

摘要/Abstract

摘要： 【目的】探究单点氨基酸替换对杂合肽KL-21溶血活性的影响，对比氨基酸替换前后多肽结构与活性变化，初步探讨影响KL-21生物活性的关键位点和因素。【方法】在杂合肽KL-21基础上，通过生物信息学方法筛选，将其第14位甘氨酸替换为丙氨酸获得新衍生肽KL-21a。利用多种在线工具对2种多肽进行生物信息学分析，用圆二色光谱法测定多肽的二级结构，通过检测最小抑菌浓度(MIC)和最小杀菌浓度(MBC)探究多肽的抗菌活性，采用分光光度法测定多肽的溶血活性。【结果】多肽KL-21a与KL-21的大多数理化特性差异较小，但与疏水性相关的理化特性差异较明显。2种多肽的二级结构均以α-螺旋为主，KL-21a的侧链更加收缩且螺旋含量低于KL-21；二者对大肠杆菌、铜绿假单胞菌、金黄色葡萄球菌和白色假丝酵母菌均有抑杀活性，对金黄色葡萄球菌抑杀效果最为相近。当多肽浓度为256 μg/mL时，KL-21溶血率＞50%，KL-21a没有溶血活性。【结论】衍生肽KL-21a具有良好抑菌活性，且溶血作用明显降低，表明第14位氨基酸是影响KL-21生物活性的关键位点，可为后续抗菌肽设计和改造提供依据和参考。

关键词: 抗菌肽; 杂合肽; 氨基酸替换; 抗菌活性; 溶血作用

Abstract: 【Objective】 The experiment was to explore the effect of single amino acid substitution on the hemolytic activity of heterozygant peptide KL-21,compare the changes in the structure and activity of polypeptide before and after amino acid substitution,and initially explore the key sites and factors affecting the bioactivity of KL-21.【Method】 Based on the heterozygous peptide KL-21,a new derivative peptide KL-21a was obtained using bioinformatics method using replacing the 14th glycine with alanine.Bioinformatics analysis of two polypeptides was performed using a variety of online tools.The secondary structure of polypeptide was determined by circular dichroism spectroscopy.The antibacterial activity of peptides was investigated by detecting minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC).The hemolytic activity of polypeptides was determined by spectrophotometry.【Result】 Most of the physical and chemical properties of peptides KL-21a and KL-21 were less different,but the differences related to hydrophobicity were more obvious.The secondary structure of the two peptides was dominated by alpha-helix,and the side chain of KL-21a was more contracted and the helix content was lower than that of KL-21.They showed inhibitory activity against Escherichia coli,Pseudomonas aeruginosa,Staphylococcus aureus and Candida albicans,and had the most similar inhibitory effect against Staphylococcus aureus.When the concentration of polypeptide was 256 μg/mL,the hemolysis rate of KL-21 was greater than 50%,and KL-21a had no hemolysis activity.【Conclusion】 The derivative peptide KL-21a had good antibacterial activity,and the hemolysis effect was significantly reduced.The results indicated that the 14th amino acid was the key site affecting the bioactivity of KL-21,which could provide basis and reference for the subsequent design and modification of antimicrobial peptides.

Key words: antimicrobial peptides; heterozygous peptide; amino acid substitution; antimicrobial activity; hemolysis

中图分类号:

S816.7

韩晋辉, 翟培, 吕文平. 单点氨基酸替换对杂合肽KL-21生物活性的影响[J]. 中国畜牧兽医, 2025, 52(5): 2400-2408.

HAN Jinhui, ZHAI Pei, LYU Wenping. Effect of Single Amino Acid Substitution on Bioactivity of Heterozygous Peptide KL-21[J]. China Animal Husbandry and Veterinary Medicine, 2025, 52(5): 2400-2408.

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