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

Abstract

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

CLC Number:

S816.7

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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Effect of Single Amino Acid Substitution on Bioactivity of Heterozygous Peptide KL-21

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