牛血红蛋白β亚基片段衍生抗菌肽的分子设计及其抗菌活性分析

doi:10.16431/j.cnki.1671-7236.2022.09.033

Abstract

Abstract: 【Objective】 This research was aimed to obtain antimicrobial peptides with low molecular weight, low hemolysis and good stability.【Method】 One peptide (YKK-18) was selected from the amino acid sequence of bovine hemoglobin β-subunit with the guidance of the structure-activity theory of antimicrobial peptides, and three modified peptides (LJ-1, LJ-2 and DLK-3) with 18 amino acid residues were designed using YKK-18 as a template.The antimicrobial activities of four peptides were reflected by minimum inhibitory concentration (MIC), the thermal stability, acid-base stability, salt ion stability and repeated freeze-thaw stability of peptides with antimicrobial activities were analyzed by agarose diffusion method, and the hemolysis of peptides with antimicrobial activities was determined by spectrophotometry.【Result】 The four designed peptides were all positively charged hydrophilic peptides, and the similarity of amino acid sequence were less than 50% compared with antimicrobial peptides in APD3 database.Secondary structure of four designed peptides had high content of alpha helix.The amphiphilic degree of modified peptides were high than template peptide.The template peptide YKK-18 had no antimicrobial activity against all tested strains, while three modified peptides had different degrees of antimicrobial activity against Escherichia coli, Salmonella, Pseudomonas aeruginosa, Staphylococcus and Candida albicans. DLK-3 peptide and LJ-1 peptide had better antimicrobial activity than LJ-2. But four designed peptides had no antimicrobial activity against Proteus mirabilis.Under the conditions of high temperature of 100 ℃, pH 4.0 to 10.0, physiological concentration of salt ions (Na⁺, K⁺, Ca²⁺, Mg²⁺, Fe³⁺, Cu²⁺) and 14 times of repeated freezing and thawing, three modified peptides had good antimicrobial activity.The hemolysis of DLK-3 was dose-dependent, and at 200 μg/mL the hemolytic rate was over 50%.However, the hemolytic rates of LJ-1 and LJ-2 were still lower than 5% at high concentration (200 μg/mL).【Conclusion】 These findings would provide a reference for molecular design and modification of antimicrobial peptides, and the obtained antimicrobial peptides LJ-1 and LJ-2 would have the potential to be used as substitutes for antibiotics.

Key words: bovine hemoglobin; antimicrobial peptide; molecular design; antimicrobial activity; stability; hemolysis

CLC Number:

S859.79

DENG Li, LI Jie, LIU Baoguo, HANG Bolin. Molecular Design and Antimicrobial Activity Analysis of Antimicrobial Peptides Derived from Bovine Hemoglobin β-subunit[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(9): 3589-3598.

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Molecular Design and Antimicrobial Activity Analysis of Antimicrobial Peptides Derived from Bovine Hemoglobin β-subunit

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