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没食子酸表没食子儿茶素和茶黄素-3,3'-二没食子酸酯对产气荚膜梭菌的抗菌作用机制。

Mechanism for antibacterial action of epigallocatechin gallate and theaflavin-3,3'-digallate on Clostridium perfringens.

机构信息

Division of Food Science and Biotechnology, Department of Bioscience and Biotechnology, Faculty of Agriculture, Graduate School, Kyushu University, Fukuoka, Japan.

Safety Science Research, R&D, Kao Corporation, Ichikai-Machi, Tochigi, Japan.

出版信息

J Appl Microbiol. 2019 Feb;126(2):633-640. doi: 10.1111/jam.14134. Epub 2018 Nov 23.

Abstract

AIM

The purpose of this study was to clarify the mechanism of the antibacterial action of two high potential and natural food additives, epigallocatechin gallate (EGCg) and theaflavin-3,3'-digallate (TF3), on Clostridium perfringens.

METHODS AND RESULTS

Minimal inhibitory concentrations were determined by the serial dilution method. Afterwards, the cells were treated with 250 or 1000 mg l of EGCg and 125 or 500 mg l of TF3 and morphological changes were observed and cell sizes were also measured under fluorescence microscopy. Our results showed that TF3 had a twice stronger antibacterial activity than EGCg against C. perfringens. Phase-contrast and fluorescence microscopy confirmed that the bacterial cells elongated without DNA segregation and septum formation in the presence of 250 mg l EGCg. While in the higher concentration of EGCg and TF3, cell growth was suppressed. Bacterial cells reached to around 12 μm after the 24 h incubation with 250 mg l EGCg, but the cells were shorter than the control at 1000 mg l of EGCg. After washing and incubating the elongated cells in fresh medium, DNA segregated at 2 h of incubation. The average cell length decreased gradually and reached the normal size at 8 h.

CONCLUSION

It seems that EGCg at a low concentration affected the proteins involved in the septum formation, DNA segregation and cell division. Furthermore, the high concentration of EGCg and TF3 seemed to cause stronger cellular damage to C. perfringens.

SIGNIFICANCE AND IMPACT OF THE STUDY

These polyphenols are widely distributed in all higher plants especially in tea plants, and people tend to use natural food additives rather than synthetic ones. EGCg and TF3, as natural food additives, can prevent C. perfringens food poisoning along with other potential health benefits.

摘要

目的

本研究旨在阐明两种高潜力天然食品添加剂表没食子儿茶素没食子酸酯(EGCg)和茶黄素-3,3′-双没食子酸酯(TF3)对产气荚膜梭菌的抗菌作用机制。

方法和结果

采用连续稀释法测定最小抑菌浓度。然后,用 250 或 1000mg/L 的 EGCg 和 125 或 500mg/L 的 TF3 处理细胞,在荧光显微镜下观察形态变化并测量细胞大小。结果表明,TF3 对产气荚膜梭菌的抗菌活性比 EGCg 强两倍。相差和荧光显微镜证实,在 250mg/L 的 EGCg 存在下,细菌细胞伸长,没有 DNA 分离和隔膜形成。而在较高浓度的 EGCg 和 TF3 下,细菌生长受到抑制。用 250mg/L 的 EGCg 孵育 24 小时后,细菌细胞长到 12μm 左右,但在 1000mg/L 的 EGCg 时,细胞比对照短。用新鲜培养基洗涤和孵育伸长细胞后,在孵育 2 小时时 DNA 分离。平均细胞长度逐渐减小,在 8 小时时恢复正常大小。

结论

似乎低浓度的 EGCg 影响了隔膜形成、DNA 分离和细胞分裂所涉及的蛋白质。此外,EGCg 和 TF3 的高浓度似乎对产气荚膜梭菌造成更强的细胞损伤。

研究的意义和影响

这些多酚广泛分布于所有高等植物中,特别是在茶树中,人们倾向于使用天然食品添加剂而不是合成添加剂。EGCg 和 TF3 作为天然食品添加剂,除了其他潜在的健康益处外,还可以预防产气荚膜梭菌食物中毒。

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