EGCG抑制波罗的海希瓦氏菌生物被膜和腐败活性的研究

doi:10.13305/j.cnki.jts.2016.02.012

摘要/Abstract

摘要： 以海产品腐败菌波罗的海希瓦氏菌（Shewanella baltica）为对象,评价了儿茶素单体抑制生物被膜的效果,并分析效果最佳的单体对该菌生物被膜和腐败活性的影响。结果表明,5种儿茶素单体均能抑制S. baltica生物被膜形成,其中表没食子儿茶素没食子酸酯（Epigallocatechin gallate,EGCG）抑制率最高,其最小抑菌质量浓度为40 µg·mL^-1,亚抑菌质量浓度10~30 µg·mL^-1的EGCG对细菌生长无显著影响。在30 µg·mL^-1 EGCG作用下,群体感应（Quorum sensing,QS）信号分子呋喃酮酰硼酸二酯（Autoinducer-2,AI-2）、二酮哌嗪类化合物（Diketopiperazine,DKPs）cyclo-(L-Pro-L-Leu)和cyclo-(L-Pro-L-Phe)活性显著下降（P<0.05）,其抑制率分别为63.95%、27.43%和21.94%。亚抑菌浓度EGCG能显著抑制S. baltica生物被膜形成、泳动能力和蛋白酶活性（P<0.05）,呈现浓度依赖性,在30 µg·mL^-1时抑制率分别为55.18%,66.71%和46.67%。因此,EGCG在亚抑菌浓度下能够干扰S. baltica的QS系统,有效抑制生物被膜形成,减弱致腐能力,研究为EGCG作为新型的QS抑制剂调控食品腐败菌奠定基础。

关键词: 波罗的海希瓦氏菌, EGCG, 生物被膜, 腐败, 群体感应, 抑制

Abstract: The anti-biofilm activity of catechin monomers from tea polyphenols against S. baltica, as the specific spoilage organism in marine product, was investigated. Effects of catechin monomer with the highest inhibitory activity on biofilm formation and spoilage potential of S. baltica were further assessed. The results showed that all five monomer of catechins significantly repressed biofilm formation of S. baltica, in which epigallocatechin gallate (EGCG) exhibited the highest activity. The minimum inhibitory concentration (MIC) of EGCG against S. baltica was 40 µg·mL^-1, and EGCG at the sub-inhibitory concentration of 10-30 µg·mL^-1 had no effect on its growth. EGCG at 30 µg·mL^-1 inhibited significantly the activity of quorum sensing (QS) autoinducers, AI-2, cyclo-(L-Pro-L-Leu) and cyclo-(L-Pro-L-Phe) (P<0.05), with reduction by 63.95%, 27.43% and 21.94% respectively. The biofilm development, swimming motility and protease activity also decreased considerably after incubation with sub-MIC of EGCG, and exhibited a dose dependent effect, which presented the reduction by 55.18%, 66.71% and 46.67% in the presence of 30 μg·mL^-1 EGCG. Hence, our results indicated that the sub-MIC of EGCG could interfere with S. baltica QS system, and inhibit the biofilm formation and spoilage potential of S. baltica. The present study suggested the potential of EGCG as QS inhibitor in controlling food spoilage bacterial.

Key words: Shewanella baltica, EGCG, biofilm, spoilage, quorum sensing, inhibition

中图分类号:

叶晓锋, 张芳, 朱军莉, 张蕾, 谢杜梅. EGCG抑制波罗的海希瓦氏菌生物被膜和腐败活性的研究[J]. 茶叶科学, 2016, 36(2): 201-209. doi: 10.13305/j.cnki.jts.2016.02.012.

YE Xiaofeng, ZHANG Fang, ZHU Junli, ZHANG Lei, XIE Dumei. Inhibition of Biofilm Development and Spoilage Potential in Shewanella baltica by Epigallocatechin Gallate[J]. Journal of Tea Science, 2016, 36(2): 201-209. doi: 10.13305/j.cnki.jts.2016.02.012.

参考文献

[1] 张雯, 倪莉, 吕旭聪, 等. 大黄鱼鲜度评价及鲜度与细菌菌群的关系[J]. 中国食品学报, 2013, 13(7): 181-188.
[2] Vogel B, Venkateswaran K, Satomi M, et al.Identification of Shewanella baltica as the most important H₂S-producing species during iced storage of danish marine fish[J]. Applied Environmental Microbiology, 2005, 71(11): 6689-6697.
[3] Simoes M, Simoes L C, Vieira M J.Species association increases biofilm resistance to chemical and mechanical treatments[J]. Water Research, 2009, 43(1): 229-237.
[4] 邓旗, 孙力军, 王雅玲, 等. 环境条件对腐败希瓦氏菌生物被膜形成能力的影响[J]. 中国食品学报, 2013, 13(10): 43-50.
[5] Novak E A, Shao H J, Daep C A, et al.Autoinducer-2 and QseC control biofilm formation and in vivo virulence of Aggregatibacter actinomycetemcomitans[J]. Infection and Immunity, 2010, 78(7): 2919-2926.
[6] Zhu S, Wu H, Zeng M, et al.The involvement of bacterial quorum sensing in the spoilage of refrigerated Litopenaeus vannamei[J]. International Journal of Food Microbiology, 2015, 192: 26-33.
[7] Ng W L, Bassler B L.Bacterial quorum-sensing network architectures[J]. Annual Review of Genetics, 2009, 43: 197-222.
[8] Waters C M, Bassler B L.Quorum sensing: cell-to-cell communication in bacteria[J]. Annual Review of Cell and Developmental Biology, 2005, 21: 319-346.
[9] Gu Q, Fu L, Wang Y, et al.Identification and characterization of extracellular cyclic dipeptides as quorum-sensing signal molecules from Shewanella baltica, the specific spoilage organism of Pseudosciaena crocea during 4℃ storage[J]. Journal of Agriculture and Food Chemistry, 2013, 61(47): 11645-11652.
[10] Packiavathy I A, Sasikumar P, Pandian S K, et al.Prevention of quorum-sensing-mediated biofilm development and virulence factors production in Vibrio spp. by curcumin[J]. Applied Microbiology and Biotechnology, 2013, 97(23): 10177-10187.
[11] Lee K M, Kim W S, Lim J, et al.Antipathogenic properties of green tea polyphenol epigallocatechin gallate at concentrations below the MIC against enterohemorrhagic Escherichia coli O157:H7[J]. Journal of Food Protection, 2009, 72(2): 325-331.
[12] Truchado P, Tomás-Barberán F A, Larrosa M, et al. Food phytochemicals act as quorum sensing inhibitors reducing production and/or degrading autoinducers of Yersinia enterocolitica and Erwinia carotovora[J]. Food Control, 2012, 24(1): 78-85.
[13] Si W, Gong J, Tsao R, et al.Bioassay-guided purification and identification of antimicrobial components in Chinese green tea extract[J]. Journal of Chromatography A, 2006, 1125(2): 204-210.
[14] 赵丽萍, 邵宛芳. 茶叶中EGCG功效研究进展[J]. 中国农学通报, 2007, 23(7): 143-147.
[15] 王慧敏, 陆海霞, 朱军莉, 等. 茶多酚抑制腐败希瓦氏菌机理研究[J]. 茶叶科学, 2014, 34(2): 149-155.
[16] 赵二科, 朱军莉, 冯立芳, 等. 冷藏大黄鱼SSO希瓦氏菌致腐能力差异机制初探[J]. 水产学报, 2015, 39(2): 274-281.
[17] 王艳, 程慧娟, 朱玲玲, 等. 鱼腥草素钠对铜绿假单胞菌生物被膜的影响[J]. 食品科学, 2013, 34(11): 173-178.
[18] 赵二科, 黄旭镇, 朱军莉, 等. 冷藏大黄鱼腐败菌群和群体感应信号分子的动态变化规律[J]. 中国食品学报, 2015, 15(4): 175-181.
[19] Asahi Y, Noiri Y, Miura J, et al.Effects of the tea catechin epigallocatechin gallate on Porphyromonas gingivalis biofilms[J]. Journal of Applied Microbiology, 2014, 116(5): 1164-1171.
[20] 梁慧, 周明旭, 朱国强, 等. 鞭毛对F18+大肠杆菌体外生物被膜形成影响的研究[J]. 中国预防兽医学报, 2014, 36(8): 624-627.
[21] Niu C, Gilbert E.Colorimetric method for identifying plant essential oil components that affect biofilm formation and structure[J]. Applied Environmental Microbiology, 2004, 70(12): 6951-6956.
[22] Atkinson S, Chang C Y, Sockett R E, et al.Quorum sensing in Yersinia enterocolitica controls swimming and swarming motility[J]. Journal of Bacteriology, 2006, 188(4): 1451-1461.
[23] Rader B A, Wreden C, Hicks K G, et al.Helicobacter pylori perceives the quorum-sensing molecule AI-2 as a chemorepellent via the chemoreceptor TlpB[J]. Microbiology, 2011, 157(9): 2445-2455.
[24] Lokender K, Sanjay C, Rajnish K, et al.Zingerone silences quorum sensing and attenuates virulence of Pseudomonas aeruginosa[J]. Fitoterapia, 2015, 102: 84-95.