Culturation of Fermented Milk Samples Containing Probiotic and Its Identification by PCR

  • AMARILA MALIK UNIVERSITAS INDONESIA
  • INDRIANI INDRIANI UNIVERSITAS INDONESIA
  • LILIANA SARAGIH UNIVERSITAS INDONESIA
  • MAKSUM RADJI UNIVERSITAS INDONESIA

Abstract

Lactic Acid Bacteria (LAB) has long been used in fermented milk because its stability, unique taste and texture and also for its health benents that are provided by the probiotic. There are many fermented dairy products in the market nowadays that declared containing living culture or as probiotics. The aim of this study was to identify LAB from fermented milk product by Visual inspection on plate culture for their morphology, as well by molecular technique performing PCR and DNA sequencing methods. Morphology and growth incubation time of each isolate was observed. A pair of primer that is highly speciiic for LAB targeting the gene encoding for 168 rRNA, which represents sequences of region 677 to 693 of E. coli numbering has been used for PCR identiiication in this study. A total of nineteen isolates were obtained from 10 fermented milk samples that were grown on de Mann Rogosa Sharpe Agar (MRSA) media. Nine out of 19 isolates which represent each of the samples were selected. Genomic DNA extracted from those 9 isolates were used as template for PCR. The results showed that 8 amplicons were positively identitied as LAB, but only three identities indicated similar species as declared, while 5 others did not.

References

1. Savadogo A, Ouattara CAT, Savadago PW, Barro N, Ouattara AS, Traore AS. Identification of exopolysaccharides-producing lactic acid bacteria from Burkina Faso fermented milk samples. African J Biotechnol. 2004. 3(3):189-94.

2. Anonim. An introduction to probiotics. diambil dari: URL: http://nccam.nih.gov/ health/probiotics/ introductionhtm. diakses 07 Maret 2011, pk. 20.40.

3. Parvez S, Malik KA, Ah Kang S, Kim H-Y. Probiotics and their fermented food products are beneficial for health. J Appl Microbiol. 2006. 100:1171-85.

4. Ljungh A, Wadstrom T. Lactic acid bacteria as probiotics. Curr Issues Intest Microbiol. 2006. 7(2):73-89

5. Heilig HGHJ, Zoetendal EG, Vaughan EE, Marteau P, Akkermans ADL, de Vos WM. Molecular diversity of Lactobacillus sp. and other lactic acid bacteria in the human intestine as determined by specific amplification of 16S ribosomal DNA. Appl Environ Microbiol. 2002. 68:114-23. .

6. Malik A, Felicia, Radji M, Oetari A. ldentiiikasi bakteri asam laktat penghasil eksopolisakarida asal sumber lokal menggunakan gen penyandi 16S rRNA. Sains Indones. 2007. 12(2):1-6

7. De Man JC, Rogosa M, Sharpe ME. A medium for the cultivation of lactobacilli. J Appl Bacteriol.1960. 23:130-5

8. Malik A, Ariestanti DM, Nurfachtiyani A, Yanuar A. Skrining gen glukosiltransferase (gti) dari bakteri asam laktat penghasil eksopolisakarida. Makara (Seri Sains). 2008. 1221-6.

9. Malik A, Radji M, Kran S, Dijkhuizen L. Screening of lactic acid bacteria from Indonesia reveals glucansucrase and fructansucrase genes in two different Weissella confuse: strains from soya. FEMS Microbiol Lett. 2009. 300:131-8.

10. Malik A, Hermawati AK, Hestiningtyas M, SoemiatiA, Radji M. Isolasi dan skrining molekuler bakteri asam laktat pembawa gen glukansukrase dari makanan dan minuman mengandung gula. Makara (Sains). 2010. 14:57-62.

11. Geneaid. Gel/PCR DNA Fragments Extraction Kit. October, 2002.

12. Altsehul SF, Gish W, Miller W, Myers EW, Lipman DJ. Basic local alignment search tool. J Mol Biol. 1990. 215(3):403-10.

13.Weaver RF. Molecular biology. 3rd Edition. New York: McGraW Hill; 2005. 99-106

14. Mahenthiralingam E, Marchbank A, Drevinek P, Garaiova I, Plummer S. Use of colony-based bacterial strain typing for tracking the fate of Lactobacillus strains during human consumption. BMC Microbiol. ' 2009. (9):251-65.

15. Randazzo CL, Torriani S, Akkermans ADL, Vos WM de, Vaughan EE. Diversity, dynamics, and activity of bacterial communities during production of an artisanal Sicilian cheese as evaluated by 168 rRNA analysis. Appl Environ Microbiol. 2002. 68(4):1882-92.

16. Alberts B, Johnson A, Lewis J, Raff M, Roberts K, Walter P. Molecular biology of the cell, 5th, New York: Garland Science, Taylor & Francis Group. G: 15 & G: 28.
Published
2011-04-30
How to Cite
MALIK, AMARILA et al. Culturation of Fermented Milk Samples Containing Probiotic and Its Identification by PCR. JURNAL ILMU KEFARMASIAN INDONESIA, [S.l.], v. 9, n. 1, p. 40-45, apr. 2011. ISSN 2614-6495. Available at: <http://jifi.farmasi.univpancasila.ac.id/index.php/jifi/article/view/324>. Date accessed: 23 nov. 2024.
Citation Formats
Issue
Vol 9 No 1 (2011): JIFI
Section
Articles
License

Licencing
All articles in Jurnal Ilmu Kefarmasian Indonesia are an open-access article, distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License which permits unrestricted non-commercial used, distribution and reproduction in any medium.

This licence applies to Author(s) and Public Reader means that the users mays :

  • SHARE:
    copy and redistribute the article in any medium or format
  • ADAPT:
    remix, transform, and build upon the article (eg.: to produce a new research work and, possibly, a new publication)
  • ALIKE:
    If you remix, transform, or build upon the article, you must distribute your contributions under the same license as the original.
  • NO ADDITIONAL RESTRICTIONS:
    You may not apply legal terms or technological measures that legally restrict others from doing anything the license permits.

It does however mean that when you use it you must:

  • ATTRIBUTION: You must give appropriate credit to both the Author(s) and the journal, provide a link to the license, and indicate if changes were made. You may do so in any reasonable manner, but not in any way that suggests the licensor endorses you or your use.

You may not:

  • NONCOMMERCIAL: You may not use the article for commercial purposes.

Creative Commons License

This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License.