生物技术通报 ›› 2022, Vol. 38 ›› Issue (9): 147-157.doi: 10.13560/j.cnki.biotech.bull.1985.2022-0396

• 细菌耐药性专题(专题主编: 刘雅红 教授 孙坚 教授) • 上一篇    下一篇

ε-聚赖氨酸对阪崎克罗诺杆菌细胞结构与生物被膜形成的影响

石成龙1(), 汪锡武1, 李安琪1, 钱森和1,2, 王洲1,2(), 赵世光1,2, 刘艳1,2, 薛正莲1,2   

  1. 1.安徽工程大学生物与食品工程学院,芜湖 241000
    2.安徽省工业微生物分子育种工程实验室,芜湖 241000
  • 收稿日期:2022-04-01 出版日期:2022-09-26 发布日期:2022-10-11
  • 作者简介:石成龙,男,硕士研究生,研究方向:微生物制药;E-mail: 441105961@qq.com
  • 基金资助:
    国家自然科学基金项目(31772081);国家自然科学基金项目(31871781)

Effect of ε-Polylysine on the Cell Structure and Biofilm Formation of Cronobacter sakazakii

SHI Cheng-long1(), WANG Xi-wu1, LI An-qi1, QIAN Sen-he1,2, WANG Zhou1,2(), ZHAO Shi-guang1,2, LIU Yan1,2, XUE Zheng-lian1,2   

  1. 1. College of Biological and Chemical Engineering,Anhui Polytechnic University,Wuhu 241000
    2. Anhui Provincial Engineering Laboratory for Molecular Breeding of Industrial Microbial,Wuhu 241000,
  • Received:2022-04-01 Published:2022-09-26 Online:2022-10-11

摘要:

ε-聚赖氨酸(ε-polylysine,ε-PL)作为一种抗菌活性强、安全性高、热稳定性好的阳离子型天然多肽,已作为安全的食品防腐剂受到关注。以阪崎克罗诺杆菌(Cronobacter sakazakii)为供试菌株,揭示ε-PL的抑菌机理,为细菌耐药性和食品防腐措施研究提供理论支持。研究了ε-PL作用下对阪崎克罗诺杆菌的最小抑菌浓度(minimum inhibitory concentration,MIC)、细胞膜壁通透性、细胞表面疏水性及运动性等生理特性的影响,并利用透射电子显微镜对ε-PL作用下的细菌细胞形态变化进行了观察,在此基础上,探究了ε-PL对阪崎克罗诺杆菌生物被膜(biofilm,BF)的抑制和清除作用效果,并利用荧光染色显微观察清除后被膜菌通透性的改变。ε-PL对阪崎克罗诺杆菌的抑菌活性具有浓度依赖性,ε-PL对阪崎克罗诺杆菌ATCC51329的MIC为256 μg/mL。ε-PL能够增强细胞膜壁通透性,使其细胞内容物如核酸、碱性磷酸酶等大量渗出,从而表现对阪崎克罗诺杆菌的杀菌作用。同时ε-PL能够降低阪崎克罗诺杆菌的表面疏水性和运动性,进而影响阪崎克罗诺杆菌生物被膜的形成。ε-PL对阪崎克罗诺杆菌生物被膜抑制和清除的作用效果显著,结合物理振荡,可大大提高生物被膜清除效率。ε-PL能够破坏阪崎克罗诺杆菌的细胞结构,从而达到抑菌效果。ε-PL能够降低阪崎克罗诺杆菌细胞表面的疏水性、运动性,进而ε-PL能够抑制生物被膜的形成,对成熟生物被膜也具有清除作用。

关键词: ε-聚赖氨酸, 阪崎克罗诺杆菌, 细胞结构, 抑菌, 生物被膜

Abstract:

ε-polylysine(ε-PL),as a cationic natural peptide with strong antibacterial activity,high safety and good thermal stability,has been applied as a food preservative. Having Cronobacter sakazakii as the test strain,the bacteriostatic mechanism of ε-PL was revealed,which may provide theoretical support for the study of bacterial drug resistance and food preservative measures. The effects of ε-PL on the physiological characteristics of C. sakazakii,such as the minimum inhibitory concentration(MIC),membrane wall permeability,cell surface hydrophobicity and motility,were studied. The morphological changes of C. sakazakii cells treated with ε-PL were observed by transmission electron microscope,and the scavenging and inhibitory effects on the biofilm(BF)of C. sakazakii were investigated,and the permeability of the cell in the biofilm was also observed by fluorescence staining. ε-PL had positively concentration-dependent antibacterial activity against C. sakazakii,the MIC of ε-PL to C.sakazakii ATCC51329 was 256 μg/mL. Physiological studies on bacteria have found that ε-PL enhanced the permeability of cell membrane wall,and transuded cell contents such as nucleic acid and alkaline phosphatase,thus presented the bactericidal effect on C.sakazakii. Meanwhile,ε-PL reduced the surface hydrophobicity and motility of C. sakazakii,and then affected the formation of the biofilm. ε-PL demonstrated a significant effect on the inhibition and eradication of biofilm,and the removal efficiency of biofilm can be greatly improved when combined with physical oscillation. ε-PL destroyed the cell structure of C. sakazakii and the bacteriostatic effect was achieved. By reducing the cell surface hydrophobicity and motility of C. sakazakii,ε -PL also inhibited the formation of biofilm,and removed the mature biofilm.

Key words: ε-polylysine, C. sakazakii, cell structure, bacteriostasis, biofilm