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Dairy Science & Technology

, Volume 89, Issue 6, pp 531–540 | Cite as

Phenotypic and genotypic characterization of lactobacilli from Churpi cheese

  • Prashant
  • Sudhir Kumar Tomar
  • Rameshwar Singh
  • Subhash Chandra Gupta
  • Dilip K. Arora
  • Balwindar Kumar Joshi
  • Dinesh KumarEmail author
Original Article

Abstract

The aim of this study was to investigate the diversity of lactobacilli in Churpi cheese, a traditional variety of cheese made from yak milk. Thirty-five lactobacilli isolated from five different samples procured from different parts of Arunachal Pradesh (India) were analyzed by phenotypic and genotypic methods. The 16S rDNA sequencing for all the isolates was performed. Five different species of lactobacilli were isolated from Churpi cheese with the predominance of Lactobacillus paracasei followed by Lactobacillus plantarum. All the isolates were further evaluated for their technological properties such as citrate utilization, exopolysaccharide (EPS), bacteriocin, and acid production. Forty-three percent isolates were found positive for citrate utilization, while 14% were found good acid producer, and 11% for EPS production. Lactobacillus coryniformis was an uncommon species found in Churpi cheese. Isolates obtained in this study can be potentially used for the development of defined strain starter for Churpi cheese.

Churpi cheese Lactobacillus 16S rDNA yak 

Churpi

Abstract

Churpi Arunachal Pradesh 5 Churpi 35 16S rDNA Churpi 5 Lactobacillus paracasei Lactobacillus plantarum 43% 14% 11% Lactobacillus coryniformis Churpi Churpi

Churpi Lactobacillus 16S rDNA 

Caractérisation phénotypique et génotypique de lactobacilles de fromage Churpi

Résumé

Le but de cette étude était d’évaluer la diversité des lactobacilles du fromage Churpi, une variété traditionnelle de fromage fabriqué à partir de lait de yak. Trente-cinq lactobacilles isolés de cinq échantillons différents obtenus de différents endroits d’Arunachal Pradesh (Inde) ont été analysés à l’aide de méthodes phénotypiques et génotypiques. Le séquençage de l’ADNr 16S de tous les isolats a été réalisé. Cinq espèces différentes de lactobacilles ont été isolées du fromage Churpi avec une prédominance de Lactobacillus paracasei suivi par Lactobacillus plantarum. Tous les isolats ont ensuite été évalués pour leurs propriétés technologiques telles que l’utilisation du citrate, la production d’exopolysaccharides (EPS), de bactériocine et d’acide. 43 % des isolats se sont avérés positifs pour l’utilisation du citrate, 14 % bons producteurs d’acide et 11 % producteurs d’EPS. Lactobacillus coryniformis, espèce rare dans le fromage, a été trouvé dans le fromage Churpi. Les isolats obtenus dans la présente étude pourraient servir au développement de levains de souches définies pour la fabrication de fromage Churpi.

fromage Churpi Lactobacillus ADNr 16S lait de yak 

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References

  1. [1]
    Anthony E.E., A note on capsule staining, Science 73 (1931) 319.CrossRefGoogle Scholar
  2. [2]
    Ayad E.H.E., Verheul A., de Jong C., Wouters J.T.M., Smit G., Flavour forming abilities and amino acid requirements of Lactococcus lactis strains isolated from artisanal and non-dairy origin, Int. Dairy J. 9 (1999) 725–735.CrossRefGoogle Scholar
  3. [3]
    Balcazar J.L., de Blas I., Ruiz-Zarzuela I., Vendrell D., Girones O., Muzquiz J.L., Sequencing of variable regions of the 16S rRNA gene for identification of lactic acid bacteria isolated from the intestinal microbiota of healthy salmonids, Comp. Immunol. Microbiol. Infect. Dis. 30 (2007) 111–118.CrossRefGoogle Scholar
  4. [4]
    Barefoot S.F., Klaenhammer T.R., Detection and activity of lactacin B, a bacteriocin produced by Lactobacillus acidophilus, Appl. Environ. Microbiol. 45 (1983) 1808–1815.Google Scholar
  5. [5]
    Berthier F., Ehrlich S.D., Rapid species identification within two groups of closely related lactobacilli using PCR primers that target the 16S/23S rRNA spacer region, FEMS Microbiol. Lett. 161 (2006) 97–106.CrossRefGoogle Scholar
  6. [6]
    Crow V., Curry B., Hayes M., The ecology of non-starter lactic acid bacteria (NSLAB) and their use as adjuncts in New Zealand Cheddar, Int. Dairy J. 11 (2001) 275–283.CrossRefGoogle Scholar
  7. [7]
    Dubernet S., Desmasures N., Gueguen M., A PCR-based method for identification of lactobacilli at the genus level, FEMS Microbiol. Lett. 214 (2002) 271–275.CrossRefGoogle Scholar
  8. [8]
    Heilig G.H.J., Zoetendal E.G., Marteau E.E., Akkermans A.D.L., de Vos W.M., Molecular diversity of Lactobacillus spp. and other lactic acid bacteria in the human intestine as determined by specific amplification of 16S ribosomal DNA, Appl. Environ. Microbiol. 68 (2002) 114–123.CrossRefGoogle Scholar
  9. [9]
    Kempler G.M., McKay L.L., Improved medium for detection of citrate-fermenting Streptococcus lactis subsp. diacetylactis, Appl. Environ. Microbiol. 39 (1980) 926–927.Google Scholar
  10. [10]
    Kim M., Chun J., Bacterial community structure in kimchi, a Korean fermented vegetable food, as revealed by 16S rRNA gene analysis, Int. J. Food Microbiol. 103 (2005) 91–96.CrossRefGoogle Scholar
  11. [11]
    Klijn N., Weerkamp A.H., de Vos W.M., Detection and characterization of lactoseutilizing Lactococcus subsp. in natural ecosystems, Appl. Environ. Microbiol. 61 (1995) 788–792.Google Scholar
  12. [12]
    Lane D.J., 16S/23S rRNA sequencing, in: Stackebrandt E.R., Goodfellow M. (Eds.), Nucleic Acid Techniques in Bacterial Systematic, John Wiley & Sons Ltd., Chichester, UK, 1991, pp. 115–175.Google Scholar
  13. [13]
    Pospiech A., Neumann B., A versatile quickprep of genomic DNA from Gram-positive bacteria, Trends Genet. 11 (1995) 217–218.CrossRefGoogle Scholar
  14. [14]
    Sancheza I., Sesenab S., Povedaa J.M., Cabezasa L., Palopb L., Phenotypic and genotypic characterization of lactobacilli isolated from Spanish goat cheeses, Int. J. Food Microbiol. 102 (2005) 355–362.CrossRefGoogle Scholar
  15. [15]
    Stackebrandt E., Goebel B.M., Taxonomic note: a place for DNA-DNA reassociation and 16S rRNA sequence analysis in the present species definition in bacteriology, Int. J. Syst. Bacteriol. 44 (1994) 846–849.CrossRefGoogle Scholar
  16. [16]
    Tamang J.P., Sarkar P.K., Microbiology of mesu, a traditional fermented bamboo shoot product, Int. J. Food Microbiol. 29 (1996) 49–58.CrossRefGoogle Scholar
  17. [17]
    Veljovic K., Terzic-Vidojevic A., Vukasinovic M., Strahinic I., Begovic J., Lozo J., Ostojic M., Topisirovic L., Preliminary characterization of lactic acid bacteria isolated from Zlatar cheese, J. Appl. Microbiol. 103 (2007) 2142–2152.CrossRefGoogle Scholar
  18. [18]
    Ward L.J.H., Timmins M.J., Differentiation of Lactobacillus casei, Lactobacillus paracasei and Lactobacillus rhamnosus by polymerase chain reaction, Lett. Appl. Microbiol. 29 (1999) 90–92.CrossRefGoogle Scholar
  19. [19]
    Weerkamp A.H., Klijn N., Neeter R., Smit G., Properties of mesophilic lactic acid bacteria from raw milk and naturally fermented raw milk products, Neth. Milk Dairy J. 50 (1996) 319–332.Google Scholar
  20. [20]
    Wiener G.H., Jianlin H., Ruijun L., Production characteristics of yak, in: The Yak, 2nd Edn., RAP Publication, FAO Regional Office for Asia and the Pacific, Bangkok, Thailand, 2003, pp. 119–171.Google Scholar
  21. [21]
    Wu X.H., Luo Z., Yu L., Ren F.-Z., Han B.-Z., Nout M.J.R., A survey on composition and microbiota of fresh and fermented yak milk at different Tibetan altitudes, Dairy Sci. Technol. 89 (2009) 201–209.CrossRefGoogle Scholar
  22. [22]
    Young J.P.W., Downer H.L., Eardly B.D., Phylogency of the phototrophic Rhizobium strain BTAil by polymerase chain reactionbased sequencing of a 16S rRNA gene segment, J. Bacteriol. 173 (1991) 2271–2277.Google Scholar

Copyright information

© Springer S+B Media B.V. 2009

Authors and Affiliations

  • Prashant
    • 1
  • Sudhir Kumar Tomar
    • 2
  • Rameshwar Singh
    • 2
  • Subhash Chandra Gupta
    • 1
  • Dilip K. Arora
    • 3
  • Balwindar Kumar Joshi
    • 1
  • Dinesh Kumar
    • 1
    Email author
  1. 1.Genes and Genetic Resources Molecular Analysis LabNational Bureau of Animal Genetic ResourcesKarnal, HaryanaIndia
  2. 2.Dairy Microbiology DivisionNational Dairy Research InstituteKarnal, HaryanaIndia
  3. 3.National Bureau of Agriculturally Important MicroorganismsUttar PradeshIndia

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