食品科学

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His-N2蛋白体外排除抑制大肠杆菌DH5α黏附猪肠黏液蛋白的研究

都立辉,张 虹,施荣华,和肖营,刘 琴   

  1. 南京财经大学食品科学与工程学院,江苏省粮油品质控制及深加工技术重点实验室,江苏 南京 210023
  • 出版日期:2014-06-15 发布日期:2014-07-03

in vitro Exclusive Inhibition of E. coli DH5α Adhesion to Porcine Intestinal Mucus by His-N2 Protein

DU Li-hui, ZHANG Hong, SHI Rong-hua, HE Xiao-ying, LIU Qin   

  1. Jiangsu Key Laboratory of Quality Control and Further Processing of Cereals and Oils, College of Food Science and Engineering,
    Nanjing University of Finance and Economics, Nanjing 210023, China
  • Online:2014-06-15 Published:2014-07-03

摘要:

以植物乳杆菌KLDS1.0320候选表面黏附蛋白NP_785232 N端前两个结构域组成的融合表达蛋白为研究对象,采用体外模拟肠道环境的方法研究其对大肠杆菌DH5α黏附猪肠黏液的排除抑制作用。对猪肠黏液进行固定,加入His-N2融合表达蛋白37 ℃孵育1 h,再加入大肠杆菌DH5α菌悬液37 ℃孵育1 h,洗去未黏附的菌体,之后用1%Triton X-100进行裂解,将裂解液涂布LB琼脂平板以进行菌落计数。结果表明:pH值为6.6 时,相对于缓冲液阴性对照组,His-N2蛋白对大肠杆菌DH5α黏附猪肠黏液的抑制率为(50.37±3.66)%,相对于BSA蛋白对照组的抑制率为(38.33±4.55)%;pH值为7.5时,相对于缓冲液阴性对照组,His-N2蛋白对大肠杆菌DH5α黏附猪肠黏液的抑制率为(42.18±3.44)%,相对于BSA蛋白对照组的抑制率为(34.48±3.90)%。在体外模拟条件下,由植物乳杆菌KLDS1.0320的候选表面黏附蛋白NP_785232 N端前两个结构域组成的His-N2蛋白能排除抑制大肠杆菌DH5α对猪肠黏液蛋白的黏附。

关键词: 植物乳杆菌, His-N2蛋白, 大肠杆菌, 猪肠黏液, 排除抑制

Abstract:

His-N2 protein, consisting of two domains at the N terminus of Lactobacillus plantarum KLDS 1.0320 adhesion
surface protein NP_785232, was selected as the candidate in this work. This protein was used to explore the exclusive
inhibition against E. coli DH5α adhesion to porcine intestinal mucus in vitro. First, porcine mucus was fixed on 96-well cell
plates and incubated at 37 ℃ for 1 h after adding His-N2 protein, and then the solution containing E. coli DH5α was added.
After co-incubation at 37 ℃ for 1 h, the mucus was washed and disrupted with 1% Triton X-100. Ten-fold dilutions of the
cell lysates were prepared and spread on LB agar plates for counting of E.coli DH5α. Results indicated that His-N2 protein
could inhibit the adhension of E. coli DH5α to porcine intestinal mucus by (50.37 ± 3.66)% compared with PBS and by
(38.33% ± 4.55)% compared with BSA protein at pH 6.6, while the inhibitory rates were (42.18 ± 3.44)% and
(34.48 ± 3.90)% at pH 7.5, respectively. Therefore, His-N2 protein is involved in the exclusive inhibition of E. coli DH5α
adhersion to porcine intestinal mucus in vitro.

Key words: Lactobacillus plantarum, His-N2 protein, E. coli DH5α, porcine mucus, exclusive inhibition