Molecular Characterization of Lactobacillus Acidophilus by SDS-PAGE and Plasmid Analysis

Author(s)

Memoona Nosheen , Tariq Masud ,

Download Full PDF Pages: 27-35 | Views: 272 | Downloads: 105 | DOI: 10.5281/zenodo.3484477

Volume 3 - August 2019 (08)

Abstract

Lactobacillus acidophilus is well known because of its probiotic role hence gain importance in food and nutrition. Keeping in view the importance of Lactobacillus acidophilus there was dire need to identify the strain accurately. Phenotypically identified 27 strains were characterized by SDS-PAGE and plasmid profile. SDS-PAGE analysis had shown that most of the strains were strongly coupled with little variation. Only two strains do not belong to any cluster showing the authenticity of SDS-PAGE. Further Plasmid profile of DNA of lactobacillus acidophilus represent the characteristics pattern, and among 27 strains 15 isolates showed the presence of plasmid. Based on number and size of plasmid the strains are grouped into three distinct groups. A combination of SDS-PAGE and plasmid analysis has been shown a practical method to use for identification of Lactobacillus at species level. However the conventional method should be combined with genotypic method for a thorough analysis of strain

Keywords

Not Provided By Author

References

  1. Anonymous. 2004.Lactobacillus acidophilus. (http://dwb.unl.edu/teacher/nsf/c11links/www.bact.wise.eduscienceed/lactobacillusacidophilus.htm).
  2. Chou. L and B. Weimer. 1999. Isolation and characterization of acid and bile-tolerant isolates from strains of Lactobacillus acidophilus. J. Dairy Sci., 82: 23-31.
  3. Anderson, D. G. and L. L. Mckay. 1983. Simple and rapid method for isolating large plasmid DNA from Lactic Streptococci. Appl. Environ. Microbiol., 46: 549-552. .
  4. Daniel, J., O. Sullivan and T. R. Klaenhammer 1993. Rapid mini-prep isolation of high-quality plasmid DNA from Lactococcus and Lactobacillus species. Appl. Environ. Microbiol., 59 (8): 2730-2733.
  5. Davies, F. L., H. M. Underwood and M. J. Gasson 1981. The value of plasmid profiles for strain identification of Lactic acid Streptococci and the relationship between Streptococcus lactis 712, ML 3 and C2. J. Appl. Bacteriol., 51: 325-337.
  6. Felten, A., C. Barreau, C. Bizet, P.H Lagrange and A. Phillippon. 1999. Lactobacillus species identification, H2O2 production, and antibiotic resistance and correlation with human clinical status. J. Clin. Microbiol., 37 (3):729-733.
  7. Gatti, M., E. Fornasari, and E. Neviani. 1997. Cell- wall protein profiles of dairy thermophilic Lactobacilli. Lett. Appl. Microbiol., 25: 345-348.
  8. Gomez-Zavaglia, A., A. Abraham, S. Giorgeri and  G. Deantoni. 1999. Application of polyacrylamide gel electrophoresis and capillary gel electrophoresis to the analysis of Lactobacillus delbrueckii whole cell proteins. J. Dairy Sci., 82: 870-877.
  9. Josephsen, J. and E. W. Nielsen. 1988. Plasmid profiles and bactreriophage sensitivity of bacteria of cheddar starter used for five years without rotation. Milchissenschaft. Milk Sci., 43 (4): 219-223.
  10. Klaenkammer, T. R. And S. M. Sutherland. 1980. Detection of plasmid deoxyribonucleic acid in an isolate of Lactobacillus acidophilus. Appl. Environ Microbiol., 39 (3): 671-674.
  11. Klaenhammer, T. R., R. barangou, B. Logan Buck, M.  Andrea Azcarate-Peril, E. Altermann. 2005. Genomic features of Lactic acid bacteria effecting bioprocessing and health. FEMS Microbiol. Reviews. 29: 393-409.
  12. Laemmli, U. K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 227: 680-685.
  13. Leisner, J. B. Pot, H. Christensen, G. Rusul, J.E Olsen, B. W. Wee, K. Muhammad and H. M. Ghazali. 1998. Identification of Lactic acid bacteria from chili Bo, a Malaysian food ingredient. Appl. Environ. Microbiol., 65 (2): 599-605.
  14. Lin, J. H. C. And D. C. Savage. 1985. Cryptic plasmids in Lactobacillus strains isolated from the murine gastrointestinal tract. Appl. Environ. Microbiol., 49 (4): 1004-1006.
  15. McCartney, A. L. 2002. Application of molecular biological methods for studying probiotics and the gut flora. Brit. J. Nutr., 88 (1): 29-38.
  16. Moreira, J. L. S., R. M. Mota, M.F. Horta, S. M. R. Teixeira, E. Neumann, J. R. Nicoli and A. C. Nunes. 2005. Identification to the species level of Lactobacillus isolated in probiotic prospecting studies of human, animal or food origin by 16S-23S rRNA restriction   profiling. (http://www.biomedcentral.com/1471-2180/5/15).
  17. Perez, G., E. Cardell and V. Zarte. 2000. Protein fingerprinting as a complementary analysis to classical phenotyping for the identification of Lactic acid bacteria from Tenerife cheese. Lait., 80: 589-600.
  18. Pot, B., C. Hertel, W. Ludwig, P. Descheemaeker, K. Kersters and K. H. Schleifer. 1993. Identification and classification of Lactobacillus acidophilus, L. Gasseri and L. Johnsonii strains by SDS-PAGE and rRNA-targeted oligonucleotide probe hybridization. J. Gen, Microbiol., 139 (3): 513-17.
  19. Reid, G. 1999. The scientific bases for probiotic strains of Lactobacillus. Appl. Environ. Microbiol., 65: 3763-3766.
  20. Rodtong, S. and G. W. Tannock. 1993. Differentiation of Lactobacillus strains by ribotyping. Appl. Environ. Microbiol., 59: 3480-3484.
  21. Sambrook, J., E. Tsakalidou, J. Metaxopoulos and G. Kalantzopoulos. 1989. Molecular cloning, 2nd ed. CSH Laboratory press, New York.
  22. Sanders M. E. and T. R. Klaenhammer. 2001. The Scientific basis of Lactobacillus acidophilus NCFM functionality as a probiotic. J. Dairy Sci., 84: 319-331.
  23. Teanpaisan, R and G. Dahlen. 2006. Use of polymerase chain reaction techniques and sodioum dodecyl sulfate-polyacrylamide gel electrophoresis for differentiation of oral Lactobacillus species. Oral Microbiol. Immunol., 21: 79-83.
  24. Tsakalidou, W., W. Manolopoulou, E. Kabaraki, E. Zoidou, B. Pot, K. Kersters and G. Kalantzopoulos. 1994. The combined use of whole-cell protein extracts for the identification (SDS-PAGE)and enzme activity screening of Lactic acid bacteria isolated from traditional Greek dairy products. Sys.Appl. Microbiol., 17: 444-458.
  25. Tynkkynen. S., R. Satokari, M. Saarela. T. Mattila-Sandholm and M. Saxelin. 1999. Comparison of ribotyping, randomly amplified polymorphic DNA analysis, and pulsed-field gel electrophoresis in typing of Lactobacillus rhamnosus and L. Casei strains. Appl. Environ. Microbiol., 65 (9): 3908-3914.
  26. Vescovo, M., L. Morelli, and V. Bottazzi. 1982. Drug resistance plasmids in Lactobacillus acidophilus and Lactobacillus reuteri. Appl. Environ. Microbiol., 43 (1): 50-56.
  27. Wilhelm, A. H., P. Haberer, R. Geisen, J. Bjorkroth and U. Schillinger. 2001. Taxonomy and important features of probiotic microorganisms in food and nutrition. Ameri. J. Clin. Nutr., 73 (2): 365-373.
  28. Wang, R. F., W. W. Cao, and C. E. Cernigil. 1996. PCR detection and quantitatin of predominant anaerobic bacteria in human and animal fecal samples. Appl. Environ. Microbiol., 62 (4): 1242-1247
  29. Zhong, W., K. Millsap, H. B. Hobrzanska, and G. Reid. 1998. Differentiation of Lactobacillus species by molecular typing. Appl. Environ. Microbiol., 64 (7): 2418-2423.

Cite this Article: