Extraction, purification and identification of surface adhesion ligand from Lactobacillus

Xue-Yan Lin, Zhong-Hua Wang, Zhong-Xiang Niu, Jun Peng

College of Animal Science & Technology, Shandong Agriculture University, Tai’an, China.

DOI: 10.4236/abb.2012.33039                

Abstract

The objective of this experiment was to purify and analyze adhesion protein from the surface of Lactobacillus. The surface adhesion ligand of Lactobacillus was isolated with of LiCl method and (NH₄)₂SO₄ method, followed by sephadex chromatography, analyzed with discontinuous native polyacrylamide gel electrophoresis, and tested with a binding assay with intestinal epithelial cells. We get the adhesion matter by two methods: ammonium sulfate and LiCl extraction. Further purification with sephadex chromatography produced 3 components. The second component of eluted from chromatography had adhesion enhancing ability compared with the control. A single band was present in discontinuous native—poly-acrylamide gel electrophoresis analysis and molecular weight was determined to be 43-66 KDa. The purified adhesion ligand can improve the number of Lactobacillus adhering to the intestinal epithelial cells.

Keywords

Lactobacillus; Adhesion Ligand; Purify; Molecular Weight

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	Henriksson, A. and Conway, P.L. (1996) Adhesion of Lactobacillus fermentum 104-S to porcine stomach mucus. Current Microbiology, 33, 31-34. doi:10.1007/s002849900069
[2]	Zheng, Y.J., Ling, J., Wang, L.S., et al. (1999) Purification of Bacillus bifidus adhesion ligand. Journal of Microbiology and Immunology of China, 19, 196.
[3]	Turner, M.S., Peter, T. and Louisem, H. (1997) Identification and characterization of a basic cell surface-located protein from Lactobacillus fermentum BR11. Journal of Bacteriology, 179, 3310-3316.
[4]	Takahiro, T., Ritva, V., Benita, W., et al. (1995) A collagen-binding s-layer protein in Lactobacillus crispatus. Applied and Environmental Microbiology, 61, 2467-2471.
[5]	Wang, J.Z. and Fan, M. (2002) Protein technical manual. Science Publishing Company, Beijing.
[6]	Greene, J.D. and Klaenhammer, T.R. (1994) Factors involed in adherence of Lactobacilli to human CaCo-2 cells. Applied and Environmental Microbiology, 60, 4487-4494.
[7]	Avall-Jaaskelainen, S., Lindholm, A. and Palva, A. (2003) Surface display of the receptor-binding region of the Lactobacillus brevis S-Layer protein in Lactococcus lactis provides nonadhesive Lactococci with the ability to adhere to intestinal epithelial cells. Applied and Environmental Microbiology, 69, 2230-2236. doi:10.1128/AEM.69.4.2230-2236.2003
[8]	Bernet, M.F., Brassart, D. and Neeser, J.R. (1994) Lactobacillus acidophilus LA 1 binds to cultured intestinal cell Lines and inhibits cell attachment cell invasion by enterovirulen. Bacteria Gut, 35, 483-489.
[9]	Zhao, B. and He, S.J. (2002) Microbiology test. Science Publishing Company, Beijing.
[10]	Jacobs, A.A., Venema, J. and Leeven, R. (1987) Inhibition of adhesive activity of K88 fibrillae by peptides derived from the K88 adhesin. Journal of Bacteriology, 169, 735-741.
[11]	Rubinsztein-Dunlop, S., Guy, B., Lissolo, L. and Fischer, H. (2005) Identification of two new Helicobacter pylori surface proteins involved in attachment to epithelial cell lines. Journal of Medical Microbiology, 54, 427-434. doi:10.1099/jmm.0.45921-0
[12]	Shan, K. and Zhe, T. (2000) One method of quickly SDS- PAGE electrophoresis dyeing bleach which profit for protein reclaim. Progress in Biochemistry and Biophysics B, 27, 210-211.
[13]	Shandong Agriculture University (2002) Biochemistry and molecular biological test technology. Shandong Publishing Company, Tai’an