食品科学 ›› 2020, Vol. 41 ›› Issue (7): 1-7.doi: 10.7506/spkx1002-6630-20190222-143

• 基础研究 •    下一篇

乳酸对阴沟肠杆菌生物膜形成的抑制作用

刘亚文,靳盼盼,许晓曦,刘芳,孙芝兰   

  1. (1.东北农业大学食品学院,黑龙江 哈尔滨 150030;2.江苏省农业科学院农产品加工研究所,江苏 南京 210014;3.青海湖肉业有限责任公司,青海 海南州 813000)
  • 出版日期:2020-04-15 发布日期:2020-04-20
  • 基金资助:
    国家自然科学基金面上项目(31871866);青海省重点研发计划项目(2019-NK-C08); 江苏省重点研发计划(现代农业)项目(BE2017392)

Inhibitory Effect of Lactic Acid on Biofilm Formation of Enterobacter cloacae

LIU Yawen, JIN Panpan, XU Xiaoxi, LIU Fang, SUN Zhilan   

  1. (1. School of Food Science, Northeast Agricultural University, Harbin 150030, China; 2. Institute of Agricultural Products Processing, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China; 3. Qinghai Lake Meat Industry Co. Ltd., Hainanzhou 813000, China)
  • Online:2020-04-15 Published:2020-04-20

摘要: 本实验首先通过结晶紫染色来确定乳酸处理对阴沟肠杆菌CY4生物膜形成的影响,然后通过分析菌株泳动能力、胞外多糖含量、生物膜微观形态和基因表达的变化量来研究乳酸处理抑制生物膜形成的作用机制。结果表明,亚抑菌浓度的乳酸可以显著抑制菌株CY4生物膜的形成(P<0.05),显著降低CY4菌株的泳动能力(P<0.05),并且能够抑制生物膜内多糖的合成。经激光共聚焦显微镜和扫描电子显微镜观察发现,亚抑菌浓度的乳酸处理使得生物膜厚度大幅降低,且表面不平滑,孔洞增多,细菌结构松散,菌体变长,相互之间缝隙变大。实时荧光定量聚合酶链式反应结果发现,亚抑菌浓度的乳酸处理造成与菌毛和多糖合成有关基因的表达量下降。综上,亚抑菌浓度的乳酸对阴沟肠杆菌CY4生物膜的形成有明显的抑制作用,这一作用可能与乳酸干扰了菌体初期的黏附聚集能力和胞外多糖的合成分泌有关。

关键词: 阴沟肠杆菌, 生物膜, 乳酸, 胞外多糖, 细胞泳动

Abstract: The inhibitory activity of lactic acid against the biofilm formation of Enterobacter cloacae CY4 was evaluated by a crystal violet staining assay, and its potential molecular mechanism was explored by studying cell swimming and swarming motilities, exopolysaccharide production, biofilm microstructure and gene expression levels. The results showed that lactic acid at subinhibitory concentrations could significantly inhibit biofilm formation and decrease cell swimming and swarming motilities of E. cloacae CY4 (P < 0.05), as well as decrease biofilm polysaccharide production. Laser confocal microscopy and scanning electron microscopy showed that lactic acid at subinhibitory concentrations could greatly reduce the thickness of the biofilm and increase its surface roughness and porosity, as well as loosen the biofilm structure with greater cell length and wider gaps between cells. Lactic acid decreased the expression of genes involved in the synthesis of pili and polysaccharides as evaluated by real-time polymerase chain reaction (PCR). This study demonstrates that lactic acid has a significant inhibitory effect on biofilm formation of E. cloacae CY4, which may be related to its influence on cell adhesion and aggregation, and the production of extracellular polysaccharides.

Key words: Enterobacter cloacae, biofilm, lactic acid, extracellular polysaccharides, cell motility

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