Application of culture-dependent and culture-independent methods for the identification of Lactobacillus kefiranofaciens in microbial consortia present in kefir grains."

HAMET, MARÍA F; LONDERO, A.; MEDRANO MICAELA; VERCAMMEN, ELISABETH; VAN HOORDE K; GARROTE G.L.; HUYS G.; VANDAMME, PETER; ABRAHAM A.G.

FOOD MICROBIOLOGY

ACADEMIC PRESS LTD-ELSEVIER SCIENCE LTD

Lugar: Manchester; Año: 2013 vol. 36 p. 327 - 334

0740-0020

Abstract: The biological and technological characteristics of kefiran as well as it importance in grain integrity led us to analyze the kefir grain consortium focusing the search on Lactobacillus kefiranofaciens. The presence of L. kefiranofaciens in the nine kefir grains studied was demonstrated by denaturing gradient gel electrophoresis. By culture dependent methods applying a methodology focused on the search of this specie, 22 isolates with typical morphology were obtained and identified applying genotypic and phenotypic validated methods: SDS-PAGE of whole cell proteins, (GTG)5-PCR and sequence analysis of the housekeeping gene encoding the subunit of bacterial phenylalanyltRNA synthase (pheS). This polyphasic approach allowed the reliable identification of 11 L.kefiranofaciens, 5 Lactobacillus paracasei, 4 Lactobacillus kefiri and 2 Lactobacillus parakefiri. The isolation and accurate identification of L. kefiranofaciens is relevant taking into account the important role of this microorganism in grain ecosystem as well as its potential application as starter in foodfermentation.