Summary
The development of effective natural antibacterial agents is important due to the insecurity of synthetic antimicrobial agents and consumer preferences. In this study, kojic acid (KA) and tea polyphenols (TP) were found to exhibit synergistic inhibitory effect against Escherichia coli O157:H7 with a fractional inhibitory concentration index of 0.25. KA combined with TP at 25% of their minimum inhibitory concentrations (MICs) completely inactivated E. coli O157:H7 within 4 h. Subsequently, propidium iodide uptake tests, genomic DNA interaction analysis, molecular docking, field emission scanning electron microscopy and biofilm formation assay were conducted to understand the synergistic antibacterial mechanism of KA‐TP. The results demonstrated that KA‐TP attacked the cell membrane cooperatively, thus disturbing membrane integrity and cell structure, and KA could bind to the genomic DNA to affect the biofilm formation of E. coli O157:H7. Moreover, the contamination of E. coli O157:H7 on raw salmon fillets was eliminated to varying degrees after soaking treatments with KA and/or TP for 30 min. The depuration effect was further enhanced when KA combined with TP, and more than 2‐log reduction of E. coli O157:H7 was obtained under the treatment of KA‐TP at 2 MIC, suggesting that KA‐TP may serve as a natural compound disinfectant for ready‐to‐eat aquatic products.
Aims:To reveal the antibacterial mechanism of the combination of thymol and cinnamaldehyde to Listeria monocytogenes ATCC 19115 on autoclaved chicken breast.
Methods and Results:In this study, L. monocytogenes ATCC 19115 on autoclaved chicken breast was exposed to the stress of 125 μg/ml thymol and 125 μg/ml cinnamaldehyde, and transcriptome analysis was used to reveal the crucial antibacterial mechanism. According to the results, 1303 significantly differentially expressed genes (DEGs) were identified. Treated by thymol and cinnamaldehyde in combination, pyrimidine and branched-chain amino acid biosynthesis of L. monocytogenes were thwarted which impairs its nucleic acid biosynthesis and intracellular metabolism. The up-regulated DEGs involved in membrane composition and function contributed to membrane repair. Besides, pyruvate catabolism and TCA cycle were restrained which brought about the disturbance of amino acid metabolism. ABC transporters were also perturbed, for instance, the uptake of cysteine, D-methionine, and betaine was activated, while the uptake of vitamin, iron, and carnitine was repressed. Thus, L. monocytogenes tended to activate PTS, glycolysis, glycerol catabolism, and pentose phosphate pathways to obtain energy to adapt to the hostile condition. Noticeably, DEGs involved in virulence factors were totally down-regulated, including genes devoted to encoding flagella, chemotaxis, biofilm formation, internalin as well as virulence gene clusters.
Conclusions:The combination of thymol and cinnamaldehyde is effective to reduce the survival and potential virulence of L. monocytogenes on autoclaved chicken breast.
Escherichia coli O157:H7 is one of the major foodborne pathogenic bacterial that cause infectious diseases in humans. The previous found that a combination of kojic acid and tea polyphenols exhibited better activity against E. coli O157:H7 than using either alone. This study aimed to explore responses underlying the antibacterial mechanisms of kojic acid and tea polyphenols from the gene level. The functional enrichment analysis by comparing kojic acid and tea polyphenols individually or synergistically against E. coli O157:H7 found that acid resistance systems in kojic acid were activated, and the cell membrane and genomic DNA were destructed in the cells, resulting in "oxygen starvation". The oxidative stress response triggered by tea polyphenols was inhibited both sulfur uptake and the synthesis of ATP, which affected the bacteria's life metabolic process. Interestingly, we found that kojic acid combined with tea polyphenols hindered the uptake of iron that played an essential role in the synthesis of DNA, respiration, tricarboxylic acid cycle. The results suggested that the iron uptake pathways may represent a novel approach for kojic acid and tea polyphenols synergistically against E. coli O157:H7 and provided a theoretical basis for bacterial pathogen control in the food industry.
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