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

doi:10.13560/j.cnki.biotech.bull.1985.2022-0396

Abstract

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

SHI Cheng-long, WANG Xi-wu, LI An-qi, QIAN Sen-he, WANG Zhou, ZHAO Shi-guang, LIU Yan, XUE Zheng-lian. Effect of ε-Polylysine on the Cell Structure and Biofilm Formation of Cronobacter sakazakii[J]. Biotechnology Bulletin, 2022, 38(9): 147-157.

Figures/Tables 8

Fig. 1 Bacteriostatic evaluation of ε-PL against C. sakazakii A：Determination of minimum inhibitory concentration; B：growth curve. The error line in the figure refers to the standard deviation,* indicates that there is significant difference compared with CK group（P<0.05）,the same below

Fig. 2 Effect of ε-PL on membrane wall properties of C. sakazakii A：Nucleic acid UV absorption. B：AKP concentration

Fig. 3 Transmission electron microscope image of C. saka-zakii after ε-PL treatment（×15.0 k） A：Control group without treatment. B：1×MIC treatment group. C：2×MIC treatment group

Fig. 4 Effect of ε-PL on the surface hydrophobicity of C. sakazakii

Fig. 5 Effect of ε-PL on the motility of C. sakazakii

Fig. 6 Effect of ε-PL on the amount of C. sakazakii biofilm formation

Fig. 7 Effect of ε-PL on the biofilm clearance of C. sakazakii A：Static treatment. B：Shaking treatment

Fig. 8 Effects of different treatment methods on the living of biofilms（×200） A：Control group with 20 h rest. B：1×MIC group with 20 h rest. C：2×MIC group with 20 h rest. D：Control group with oscillation for 3 h. E：1×MIC group with oscillation for 3 h. F：2×MIC group with oscillation for 3 h

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