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Jang, Hyun, Oh, Choi, Kim, Yeo, Han, and Kim: Adhesion Activity of Lactobacillus plantarum PM 008 Isolated from Kimchi on the Intestine of Mice
Original Article

J Bacteriol Virol 2011;41(2):83-90.

Published online: 18 January 2011

DOI: https://doi.org/10.4167/jbv.2011.41.2.83

Adhesion Activity of Lactobacillus plantarum PM 008 Isolated from Kimchi on the Intestine of Mice

Se-Eun Jang1, Yang-Jin Hyun2, Young-Joo Oh3, Kum Boo Choi3, Taesok Kim3, Ik Hyun Yeo3, Myung Joo Han1, ∗, Dong-Hyun Kim2, ∗

1Department of Food and Nutrition, Kyung Hee University, Seoul, Korea

2Department of Life and Pharmaceutical Sciences, Kyung Hee University, Seoul, Korea

3Pulmuone Co., Seoul, Korea

∗Corresponding author: Dong-Hyun Kim, Ph.D. Department of Life and Nanopharmaceutical Sciences and Department of Pharmaceutical Science, Kyung Hee University, 1, Hoegi, Dongdaemun-ku, Seoul 130-701, Korea. Phone: +82-2-961-0374, Fax: +82-2-957-5030 e-mail: dhkim@khu.ac.kr

∗Corresponding author: Myung Joo Han, Ph.D. Department of Food and Nutrition, Kyung Hee University, 1, Hoegi, Dongdaemun-ku, Seoul 130-701, Korea. Phone: +82-2-961-0553, Fax: +82-2-968-0260 e-mail: mjhan@khu.ac.kr

      Revised 17 February 201122 March 2011       Accepted 31 March 2011

Copyright © 2011 Korean Society for Legal Medicine

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Lactic acid bacteria (LAB), including L. plantarum isolated from Kimchi, are beneficial and safe microorganisms that improve disturbances of the indigenous microflora and the host's immune system. The adhesion abilities of Kimchi-derived L. plantarum PM008 and yogurt-derived L. casei were measured in vitro and in vivo. When L. plantarum or L. casei was incubated with Caco-2 cells, these Lactobacillus strains were potently attached. When these strains were orally administered to mice, the LABs were attached on the large intestine of mice. The attachment of L. plantarum on murine intestine or Caco-2 intestinal epithelial cell lines was more potent than that of L. casei, although numbers of LAB between their feces were not different. Treatment with either L. plantarum or L. casei for 14 days suppressed fecal β-glucuronidase activity, although treatment for one day did not affect it. L. plantarum showed more potent inhibition than L. casei. In addition, L. plantarum and L. casei were stable to artificial gastric and intestinal juice. L. plantarum was more stable than L. casei. Based on these findings, the survival and adhesion effects of orally administered LAB strains in the intestine may increase numbers of LAB in intestine and express their biological activities.

Keywords: Lactic acid bacteria, Lactobacillus plantarum, Lactobacillus casei, Adhesion activity

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Figure 1.
Adhesion ability of lactic acid bacteria to Caco-2 cells. Lactic acid bacteria (closed circle, 1 × 107 L. plantarum per well; open circle, 1.0 × 107 L. casei per well) were incubated with Caco-2 cells for 2 h, washed with PBS and then inoculated in MRS agar plate.
jbv-41-83f1.tif
Figure 2.
Number of intestinal bacteria attached in colon membrane in Lactobacillus plantarum (A) or Lactobacillus casei (B)-treated mice by a fluorescent hybridization method. Lactic acid bacteria were orally administered once a day for one (a) or 14 days (b): LP1, treated with 1.0 × 109 L. plantarum per mouse for one day; LP14, 1.0 × 109 treated with L. plantarum per mouse for 14 days; LC1, 1.0 × 109 L. casei per mouse for one day; LC14, 1.0 × 109 treated with L. casei per mouse for 14 days). All values are mean ± SD. (n = 6). ∗significantly different compared to control group.
jbv-41-83f2.tif
Figure 3.
Number of intestinal bacteria attached in colon membrane in Lactobacillus plantarum (LP) or Lactobacillus casei (LC)-treated mice by a slot blot hybridization method. Lactic acid bacteria were orally administered once a day for one or 14 days: LP1, treated with 1.0 × 109 L. plantarum per mouse for one day; LP14, 1.0 × 109 treated with L. plantarum per mouse for 14 days; LC1, 1.0 × 109 L. casei per mouse for one day; LC14, 1.0 × 109 treated with L. casei per mouse for 14 days). All values are mean ± SD. (n=6). ∗significantly different compared to control group.
jbv-41-83f3.tif
Figure 4.
Effect of lactic acid bacteria on fecal bacterial. β-glucosidase (A) and β-glucuronidase activities (B) in mice. Lactic acid bacteria (LP0.2, 0.2 × 109 L. plantarum per mouse; LP1.0, 1.0 × 109 L. plantarum per mouse; LC0.2, 0.2 × 109 L. casei per mouse; LC1.0, 1 × 109 L. casei per mouse) were orally administered once a day for one (upper) or 14 days (bottom). All values are mean ± SD. (n=6). ∗significantly different compared to control group.
jbv-41-83f4.tif
Figure 5.
Stability of lactic acid bacteria against artificial gastric juice and intestinal juice. Lactic acid bacteria (L. plantarum or L. casei) were incubated in saline (white bar), artificial gastric [gray bar, pepsin (1,000 units)-contained MRS broth adjusted at pH 2.5 with 5 N HCl] or intestinal juice [black bar, GAM broth containing 0.1% mucin, 0.04% pancreatin, 0.2% bile, 0.85% NaCl, and 0.04% trypsin (pH 6.8)] for 3 h at 37°C and number of survival LAB were determined by plating serial dilutions (in PBS, pH 7.2) on MRS agar followed by incubation at 37°C for 48 h. All values are mean ± SD. (n=6). ∗significantly different compared to saline control group.
jbv-41-83f5.tif
Table 1.
Colony forming unit (CFU) of fecal bacteria grown in MRS and BL agar plates in mice orally treated with lactic acid bacteria
  Dosage Treated period (day) MRS BL
Before After Before After
L. plantarum 0.2 × 109 CFU 1 0.4 × 109 0.1 × 109 1.5 × 108 1.3 × 108
  1 × 109 CFU 1 1.6 × 109 1.5 × 109 1.1 × 108 1.0 × 108
L. casei 0.2 × 109 CFU 1 2.1 × 109 1.3 × 109 2.8 × 108 0.5 × 108
  1 × 109 CFU 1 4.1 × 109 3.8 × 109 3.8 × 108 3.7 × 108
L. plantarum 0.2 × 109 CFU 14 2.4 × 108 3.1 × 108 0.8 × 108 1.1 × 108
  1 × 109 CFU 14 2.3 × 108 3.5 × 108 0.3 × 108 1.3 × 108
L. casei 0.2 × 109 CFU 14 1.4 × 108 2.9 × 108 0.5 × 108 0.2 × 108
  1 × 109 CFU 14 2.2 × 108 3.1 × 108 0.3 × 108 3.2 × 108

Lactic acid bacteria (L. plantarum or L. casei) were orally administered once a day for one or 14 days. Numbers of intestinal bacteria were counted by plating serial dilutions (in diluted anaerobic broth, pH 7.2) on MRS and BL agars followed by anaerobic incubation at 37°C for 48 h. All values are mean (n=3).

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