IV International Symposium on Lactic Acid Bacteria: Food, Health and Applications

A7 GENOTYPIC AND PHENOTYPIC CHARACTERIZATION OF 

EXOPOLYSACCHARIDE‐PRODUCING Lactobacillus STRAINS 
ISOLATED FROM KEFIR GRAINS
M.V. Gangoiti1, M.F. Hamet2, M. Medrano2, A.I. Puertas3, J, Piermaria, M.T. Dueñas3, A.G Abraham1,2

1AreaBioquímica y Control de Alimentos – Facultad de ciencias Exactas, UNLP. 47 y 115 (1900) La Plata, Buenos Aires,
Argentina. 2Centro de Investigación y Desarrollo en Criotecnología de Alimentos (CIDCA). 47 y 116 (1900) La Plata,
Buenos Aires, Argentina. 3Dpto. de Química Aplicada, Dpto. de Ciencia y Tecnología de Polímeros, Facultad de Ciencias
Químicas. Universidad del País Vasco. Paseo Manuel de Lardizabal, 3 (20018), San Sebastián, País Vasco. E‐mail:
aga@biol.unlp.edu.ar

Some lactic acid bacteria (LAB) isolated from kefir grains, exert the ability to develop ropy colonies
when growing in MRS‐Agar, that could be associated to exopolysaccharide (EPS) producing
bacteria. EPS producing LAB have the ability to synthesize in situ these biopolymers, which could
act as natural thickening agents, improving texture on fermented products. This ability, and the
GRAS status that LAB possesses, confers these microorganisms an additional value to be used on
dairy industry. The aim of the present study was to conduct a genotypic and phenotypic
characterization of the EPS production of 3 Lactobacillus paracasei strains (CIDCA 8339, CIDCA
83123, CIDCA 83124) and 2 L. kefiranofaciens strains (CIDCA 83118, CIDCA 83119) isolated from
kefir grains. For genotypic characterization, the presence of the priming enzyme glycosyl
transferase (pGT) ‐essential for heteropolysaccharides synthesis‐ was determined by PCR. The
obtained products were amplified and sequenced, finding that the pGT gen was present in all the
studied strains. For phenotypic characterization, all the microorganisms were grown in milk under
their respective optimal culture conditions, and EPS production was characterized. EPS produced by
these strains was between 140 and 301,9 mg/l. Molecular weight of EPSs was determined by high
pressure liquid chromatography (HPLC) using a molecular exclusion column associated to a
refraction index detector. Different fractions were found, with molecular weights between 104‐106
Da for EPSs produced by L. paracasei and 102‐105 Da for those produced by L. kefiranofaciens. To
conduct all the determinations, two EPS producing strains were used as positive controls: L.
delbrueckii subsp bulgaricus CIDCA 332 and L. kefiranofaciens subsp kefiranofaciens JCM 6985. The
evaluation of the flow curves of the fermented products of all the studied strains showed a
pseudoplastic fluid behavior, and presented apparent viscosity values and hysteresis areas higher to
acid milk gels obtained by chemical acidification of milk with δ‐gluconolactona. It can be concluded
that Lactobacillus studied possess the gen encoding enzyme responsible for heteropolysaccharide
synthesis. The phenotypic expression of this gen results in the production of EPSs that modify
rheological behavior of fermented milks. Taking into account the present results these strains and
the EPSs they produce are interesting candidates for their application in food industry.

San Miguel de Tucumán, Tucumán, ARGENTINA. October 16-18, 2013