食品科学 ›› 2010, Vol. 31 ›› Issue (13): 148-153.doi: 10.7506/spkx1002-6630-201013035

• 基础研究 • 上一篇    下一篇

壳聚糖对革兰氏阴性菌抑菌机理的初步研究

李小芳,冯小强,杨 声*   

  1. 天水师范学院生命科学与化学学院
  • 收稿日期:2009-04-07 修回日期:2009-12-30 出版日期:2010-07-01 发布日期:2010-12-29
  • 通讯作者: 杨 声 E-mail:fengxiaoq04@lzu.cn

A Mechanism of Antibacterial Activity of Chitosan against  Gram-negative Bacteria

LI Xiao-fang FENG Xiao-qiang YANG Sheng *   

  1. College of Biology and Chemistry, Tianshui Normal University, Tianshui 741001, China
  • Received:2009-04-07 Revised:2009-12-30 Online:2010-07-01 Published:2010-12-29
  • Contact: YANG Sheng E-mail:fengxiaoq04@lzu.cn

摘要:

体外抑菌法研究壳聚糖对绿脓杆菌、奇异变形杆菌和大肠杆菌的抑菌性能。同时,对细菌表面的亲水性和负电荷进行研究,以阐述革兰氏阴性菌表面性质与壳聚糖对其抑菌性能之间的关系。以大肠杆菌作为代表性菌株,考察了壳聚糖对革兰氏阴性菌的抑菌机理。通过测定菌液中加入疏水性荧光探剂1-N-苯萘胺(NPN)后荧光强度的改变,考察壳聚糖对外膜渗透性的影响。此外,采用红外吸收光谱(FT-IR),对壳聚糖和大肠杆菌的作用产物进行表征。结果表明:革兰氏阴性菌表面的亲水性越好,所带的负电荷越多,壳聚糖表现出较好的抑菌性能;壳聚糖可增加细胞外膜的渗透性,壳聚糖与细胞膜间发生静电作用而使细胞膜破坏,最终导致菌体的死亡。

关键词: 壳聚糖, 革兰氏阴性菌, 抑菌, 机理

Abstract:

The antibacterial activities of chitosan against Pseudomonas aeruginosa (P. aeruginosa), Proteus mirabilis (P. mirabilis) and Escherichia coli (E. coli) were evaluated by optical density method. Meanwhile, bacterial cell surface hydrophilicity and negative charge analysis were investigated to illustrate the relationship between surface characteristics of Gram-negative bacterial cell wall and antibacterial activity of chitosan. To investigate the action mode of chitosan towards Gram-negative bacterial, E. coli was selected to be representative of bacteria. The permeability of the outer membrane was investigated by determining the changes in the fluorescence intensity of cells treated with the fluorescent probe of 1-N-phenylnaphthylamine (NPN). Furthermore, the interaction of chitosan with E. coli was studied by Fourier-transform infrared spectroscopy (FT-IR). Results showed that the Gram-negative bacteria with better hydrophilicity and more negatively charged cell surfaces exhibited greater interaction with chitosan and accordingly chitosan had preferable antibacterial activity against them. Chitosan increased the permeability of outer membrane and ultimately disrupted bacterial cell membranes. This damage was likely caused by the electrostatic interaction between —NH3+ groups of chitosan and carbonyl or phosphoryl groups of phospholipid components of cell membranes.

Key words: chitosan, Gram-negative bacteria, antibacterial activity, mechanism

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