Skip to main content
Log in

Specific enumeration of lactic acid bacteria in ragi tape by colony hybridization with specific oligonucleotide probes

  • Published:
World Journal of Microbiology and Biotechnology Aims and scope Submit manuscript

Abstract

Lactic acid bacteria (LAB) are among the integral microflora of ragi tape, a dry starter of Balinese rice wine, brem. The species diversity and population level of LAB present in different types of ragi tape were studied by colony hybridization using 16S and 23S rDNA targeted oligonucleotide probes. These probes were DB6, Lbc, Wgp, and Rpt, which were specific for Enterococcus faecium, Lactobacillus curvatus, Weissella spp., and Pediococcus pentosaceus, respectively. Results revealed that P. pentosaceus and Weissella spp. were the predominant LAB in ragi tape, whereas L. curvatus and E. faecium were associated specific to types of ragi tape. A 21-mer species-specific oligonucleotide probe, Rpt, that targets the 16S rDNA of P. pentosaceus was developed in this study and found to be highly specific to be used as an effective tool to enumerate population of this species in ragi tape and its population changes during rice wine production. It was detected that LAB showed active growth during the early stage of brem fermentation. A succession of growth of LAB population during the fermentation was observed in which the heterofermentative LAB, Weissella spp., grew first, followed by the proliferation of P. pentosaceus.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Alm, E.W., Oerther, D.B., Larsen, N., Stahl, D.A. & Raskin, L. 1996 The oligonucleotide probe database. Applied and Environmental Microbiology 62, 3557–3559.

    Google Scholar 

  • Ardana, M.M. & Fleet, G.H. 1989 The microbial ecology of tape ketan fermentation. International Journal of Food Microbiology 9, 157–165.

    Google Scholar 

  • Betzl, D., Ludwig, W. & Schleifer, K.H. 1990 Identification of Lactococci and Enterococci by colony hybridization with 23S rRNA-targeted oligonucleotide probes. Applied and Environmental Microbiology 56, 2927–2929.

    Google Scholar 

  • Collins, M.D., Williams, A.M. & Wallbanks, S. 1990 The phylogeny of Aerococcus and Pediococcus determined by 16S rRNA sequence analysis: description of Tetragenococcus gen. nov. FEMS Microbiology Letters 70, 255–262.

    Google Scholar 

  • Collins, M.D., Samelis, J., Metaxopoulos, J. & Wallbanks, S. 1993 Taxonomic studies on some Leuconostoc-like organisms from fermented sausages: description of a new genus Weissella for the Leuconostoc paramesenteroides group of species. Journal of Applied Bacteriology 75, 595–603.

    Google Scholar 

  • Cronk, T.C., Mattick, L.R., Steinkraus, K.H. & Hacker, L.R. 1979 Production of higher alcohols during Indonesian tape ketan fermentation. Applied and Environmental Microbiology 37, 892–896.

    Google Scholar 

  • Djien, K.S. 1972 Tape fermentation. Applied Microbiology 23, 976–978.

    Google Scholar 

  • Hadisepoetro, E.S.S., Takada, N. & Oshima, Y. 1979 Microflora in ragi and usar. Journal of Fermentation Technology 57, 251–259.

    Google Scholar 

  • Hertel, C., Ludwig, W., Obst, M., Vodgel, R.R., Hammes, W.P. & Schleifer, K.H. 1991 23 S rRNA-targeted oligonucleotide probes for the rapid identification of meat lactobacilli. Systematic and Applied Microbiology 14, 173–177.

    Google Scholar 

  • Hesseltine, C.W. & Ray, M.L. 1988 Lactic acid bacteria in muncha and ragi. Journal of Applied Bacteriology 64, 395–401.

    Google Scholar 

  • Ishimaru, Y. & Nakano, M. 1960 Studies on the microorganisms in the ragi. Report of Food Research Institute 18, 158–163 (in Japanese).

    Google Scholar 

  • Kaufmann, P., Pfefferkorn, A., Teuber, M. & Meile, L. 1997 Identification and quantification of Bifidobacterium species isolated from food with genus-specific 16S rRNA-targeted probes by colony hybridization and PCR. Applied and Environmental Microbiology 63, 1268–1273.

    Google Scholar 

  • Kozaki, M., Uchimura, Y. & Okada, S. 1992 Manual for Isolation and Identification of Lactic Acid Bacteria. Tokyo: Asakura Shoten. ISBN 4-254-43049-3 (in Japanese).

    Google Scholar 

  • Lucia, S.H., Sloan, G.L. & Heath, H.E. 1986 A simple and generally applicable procedure for releasing DNA from bacterial cells. Applied and Environmental Microbiology 51, 1138–1140.

    Google Scholar 

  • Mora, D., Fortina, M.G., Parini, C. & Manachini, P.L. 1997 Identification of Pediococcus acidilactici and Pediococcus pentosaceus based on 16S rRNA and ldhD gene-targeted multiplex PCR analysis. FEMS Microbiology Letters 151, 231–236.

    Google Scholar 

  • Neef, J.M., Van de Peer, Y., Hendriks, L. & Watcher, D.R. 1990 Compilation of small ribosomal subunit RNA sequences. Nucleic Acids Research (supplement) 18, 2237–2317.

    Google Scholar 

  • Sakurai, H., Yokota, A. & Tomita, F. 1997 Molecular cloning of an inulin fructotransferase (depolymerizing) gene from Arthrobacter sp. H-657 and its expression in Escherichia coli. Bioscience Biotechnology and Biochemistry 61, 87–92.

    Google Scholar 

  • Salama, M., Sandine, W. & Giovannoni, S.J. 1991 Development and application of oligonucleotide probes for identification of Lactococcus lactis subsp. cremoris. Applied and Environmental Microbiology 57, 1313–1318.

    Google Scholar 

  • Schleifer, K.H., Ehrmann, M., Beimfohr, M., Brockmann, E., Ludwig, W. & Amann, R. 1995 Application of molecular methods for the classification and identification of lactic acid bacteria. International Dairy Journal 5, 1081–1094.

    Google Scholar 

  • Simpson, W.J. & Taguchi, H. 1995 The genus Pediococcus with notes on the genera Tetragenococcus and Aerococcus. In The Genera of Lactic Acid Bacteria, eds. Wood, B.J.B. & Holzapfel, W.H. pp. 125–172. London: Blackie Academic & Professional. ISBN 0-7514-0215-X.

    Google Scholar 

  • Steinkraus, K.H. 1996 Indigenous fermented foods in which ethanol is a major product. In Handbook of Indigenous Fermented Foods, 2nd edn, ed. Steinkraus, K.H. pp. 363–508. New York: Marcel Dekker, Inc. ISBN 0-8247-9352-8.

    Google Scholar 

  • Sujaya, I.N., Amachi, S., Yokota, A., Asano, K. & Tomita, F. 2001a Identification and characterization of lactic acid bacteria in ragi tape. World Journal of Microbiology and Biotechnology 17, 349–357.

    Google Scholar 

  • Sujaya, I.N., Sone, T., Yokota, A., Asano, K. & Tomita, F. 2001b Studies on lactic acid bacteria in brem fermentation. Abstract on Annual Meeting of Japan Society for Bioscience, Biotechnology, and Agrochemistry. Nippon Nōgeikagaku Kaishi 75, 339.

    Google Scholar 

  • Wood, B.J.B. & Hodge, M.M. 1985 Yeast-lactic acid bacteria interactions and their contribution to fermented foodstuffs. In Microbiology of Fermented Foods, vol. I, ed. Wood, B.J.B. pp. 263–294. London: Elsevier Applied Science Publishers. ISBN 0-85334-332-1.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Sujaya, I., Amachi, S., Saito, K. et al. Specific enumeration of lactic acid bacteria in ragi tape by colony hybridization with specific oligonucleotide probes. World Journal of Microbiology and Biotechnology 18, 263–270 (2002). https://doi.org/10.1023/A:1014964613329

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1014964613329

Navigation