不同疏水性氨基酸对α-螺旋抗菌肽生物学活性的影响

doi:10.11843/j.issn.0366-6964.2019.04.012

Abstract

Abstract:

Natural antimicrobial peptides have a strong bactericidal ability, however, low cell selectivity of natural antimicrobial peptides due to their strong cytotoxicity. In order to improve the specificity of antibacterial peptides, the effects of different hydrophobic amino acids on the biological activity of α-helix antimicrobial peptides and bacteriostatic mechanisms were investigated. We employed hydrophobic amino acid tryptophan (Try, T), phenylalanine (Phe, P), valine (Val, V), alanine (Ala, A), leucine (Leu, L) and isoleucine (Ile, I) to replace X position of GRX₂RX₃RX₂RG template. Our study evaluated the secondary structure, hemolytic activity, bacteriostatic activity, salt resistance and studied bacteriostatic mechanism of peptides. The results of CD spectroscopy showed that GF, GI, GA and GL displayed typical α helix structure in the simulated environment of cell membrane, while only GV exhibited α helix structure in TFE. Hemolysis test showed that GV and GA did not exhibit hemolytic activity in the concentration of 128 μmol·L^-1, while other peptides showed high hemolytic activity. The geometric average value of minimal inhibitory concentration (MIC) of GV which has the highest therapeutic index is 3.7 μmol·L^-1. The stability test showed that GV had the stable antibacterial activity against E. coli ATCC 25922 in the presence of NH₄⁺, Zn²⁺ and Fe³⁺. The antimicrobial mechanism of antimicrobial peptide GV was further analyzed by scanning electron microscopy and inner membrane permeability test. The results showed that GV induced outflow of bacterial content via penetrating membrane of E. coli ATCC 25922 and Staphylococcus aureus ATCC 29213 and penetrated inner membrane of E. coli ATCC 25922 by time-and dose-dependence. Collectivity, the Val-rich antimicrobial peptide GV had the highest cell selectivity, and is potential for development of highly effective antibacterial drugs.

Key words: α-helical antibacterial peptides, cell selectivity, bactericidal mechanism, hemolysis, hydrophobic

CLC Number:

S859.796

XU Xinyao, DONG Na, LI Xinran, YANG Yang, WANG Zhihua, SHAN Anshan. Effect of Different Hydrophobic Amino Acids on Biological Activity of Alpha Helix Antimicrobial Peptides[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(4): 791-801.

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Effect of Different Hydrophobic Amino Acids on Biological Activity of Alpha Helix Antimicrobial Peptides

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[1]	XU Xinyao, DONG Na, LI Xinran, YANG Yang, WANG Zhihua, SHAN Anshan. Effect of Different Hydrophobic Amino Acids on Biological Activity of Alpha Helix Antimicrobial Peptides [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(4): 791-801.
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