抗菌肽LL-1对肺炎克雷伯菌ATCC 700603的抗菌机制
Antimicrobial Mechanism of Antimicrobial Peptide LL-1 Against Klebsiella pneumoniae ATCC 700603
连凯琪 1王宇航 2曾雅静 2周玲玲 3张元臣 4张明亮 3张改平 5王双山3
作者信息
- 1. 安阳工学院生物与食品工程学院,安阳 455000;河南农业大学动物医学院,郑州 450046;河南省兽用生物制品研发与应用国际联合实验室,安阳 455000
- 2. 安阳工学院生物与食品工程学院,安阳 455000
- 3. 安阳工学院生物与食品工程学院,安阳 455000;河南省兽用生物制品研发与应用国际联合实验室,安阳 455000
- 4. 河南省太行山林业有害生物野外科学观测研究站,林州 456550
- 5. 河南农业大学动物医学院,郑州 450046
- 折叠
摘要
[目的]探明新抗菌肽LL-1对肺炎克雷伯菌(Klebsiella pneumoniae)的抗菌效果和机制.[方法]测定抗菌肽LL-1对肺炎克雷伯菌的最小抑菌浓度(minimum inhibitory concentration,MIC),通过抑菌曲线评价抗菌肱LL-1对肺炎克雷伯菌的体外抗菌效果;通过检测碱性磷酸酶(ALP)、β-半乳糖苷酶(β-galactosidase)、核酸、蛋白质的泄露,评价抗菌肽LL-1对肺炎克雷伯菌细胞膜和细胞壁的影响;通过核酸凝胶电泳检测抗菌肽LL-1与肺炎克雷伯菌DNA的结合作用;通过扫描电子显微镜观察抗菌肽LL-1对肺炎克雷伯菌菌体形态的影响.[结果]抗菌肽LL-1对肺炎克雷伯菌具有良好的抗菌活性,MIC为50 μg/mL;抑菌曲线表明,抗菌肱LL-1对肺炎克雷伯菌具有生长抑制作用,且2 MIC的抗菌肽LL-1在6 h内能完全抑制肺炎克雷伯菌;抗菌肽LL-1能导致肺炎克雷伯菌的ALP和β-半乳糖苷酶发生泄漏,且1/4 MIC的抗菌肽LL-1即可导致其泄漏量极显著高于对照组(P＜0.01);核酸和蛋白质泄露的检测结果进一步证实抗菌肽LL-1能导致肺炎克雷伯菌胞内容物外流;此外抗菌肽LL-1还能与肺炎克雷伯菌DNA结合;扫描电子显微镜观察显示,抗菌肽LL-1会导致肺炎克雷伯菌表面变得粗糙、菌体变形等.[结论]抗菌肽LL-1能增加肺炎克雷伯菌细胞壁和细胞膜的通透性,并结合其DNA,从而发挥抗菌作用,本研究结果将为深入研究抗菌肽LL-1对肺炎克雷伯菌的抗菌机制奠定基础.
Abstract
[Objective]This experiment was aimed to study the antibacterial effect and mechanism of a novel antibacterial peptide LL-1.[Method]The minimum inhibitory concentration(MIC)of antibacterial peptide LL-1 against Klebsiella pneumoniae was detected,and the in vitro antibacterial effect of antibacterial peptide LL-1 against Klebsiella pneumoniae was evaluated through the antibacterial curve.Leakage of alkaline phosphatase(ALP),β-galactosidase,nucleic acid and protein were determined to evaluate the effect of antibacterial peptide LL-1 on the cell membrane and cell wall of Klebsiella pneumoniae.The binding of antibacterial peptide LL-1 to Klebsiella pneumoniae DNA was detected by nucleic acid gel electrophoresis.The effect of antibacterial peptide LL-1 on the morphology of Klebsiella pneumoniae was observed through scanning electron microscopy.[Result]Antibacterial peptide LL-1 had good antibacterial activity against Klebsiella pneumoniae,with a MIC of 50 μg/mL.The antibacterial curve showed that 2 MIC of antibacterial peptide LL-1 could completely inhibit the growth of Klebsiella pneumoniae within 6 h.Antibacterial peptide LL-1 led to the leakage of ALP and β-galactosidase of Klebsiella pneumoniae,and 1/4 MIC of antibacterial peptide LL-1 resulted in a extremely significantly higher leakage rate than the control group(P＜0.01).The detection of nucleic acid and protein leakage further confirmed that antibacterial peptide LL-1 might cause the efflux of intracellular contents in Klebsiella pneumoniae.In addition,antibacterial peptide LL-1 could bind to the DNA of Klebsiella pneumoniae.The results of scanning electron microscope observation suggested that antibacterial peptide LL-1 caused the surface of Klebsiella pneumoniae to become rough,bacterial deformation.[Conclusion]The results showed that antibacterial peptide LL-1 increased the permeability of the cell wall and membrane in Klebsiella pneumoniae,and bound with the DNA of Klebsiella pneumoniae,and thus exerted antibacterial effects.This would lay the foundation for in-depth research on the antibacterial mechanism of antibacterial peptide LL-1 against Klebsiella pneumoniae.
关键词
抗菌肽/肺炎克雷伯菌/抗菌机制/细胞壁/细胞膜Key words
antimicrobial peptide/Klebsiella pneumoniae/antimicrobial mechanism/cell wall/cell membrane引用本文复制引用
基金项目
国家自然科学基金青年项目(31802170)
河南省科技攻关计划项目(212102110090)
安阳市重点研发与推广专项(2023C01NY013)
安阳工学院博士后创新实践基地启动经费资助(BHJ2020005)
出版年
2024
国家科技期刊平台
NSTL
维普
万方数据