Morphometric parameters, zeta potential and growth rate of Lactobacillus casei Shirota by effect of different bile salts

Parámetros morfométricos, potencial zeta y tasa de crecimiento de Lactobacillus casei Shirota por efecto de diferentes sales biliares

R. González-Vázquez¹, G.F. Gutiérrez-López¹, S. Arellano-Cárdenas², E. O. López-Villegas³, D. I. Téllez-Medina¹*, and Y. Rivera-Espinoza¹

¹ Departamento de Graduados e Investigación en Alimentos, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional. Prolongación de Carpio y Plan de Ayala, C.P. 11340, México D.F. *Corresponding author. E-mail: darioiker@gmail.com Tel. 57-29-60-00, Fax 62463.

³ Departamento de Investigación, Sección de Estudios de Posgrado e Investigación, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional. Prolongación de Carpio y Plan de Ayala, C.P. 11340, México D.F.

Received December 11, 2013.
Accepted December 31, 2013.

Abstract

Effects of primary and secondary bile salts, conjugated to glycine [glycocholic (GCA) and glycodeoxycholic acid (GDCA)], taurine [taurocholic acid (TCA), taurodeoxycholic acid (TDCA)] on morphometry and growth rate of Lactobacillus casei Shirota were studied. Images obtained by scanning electron microscopy showed modification (p < 0.05) in cell minor axis and aspect ratio. The former was modified by GCA while the second by all the bile salts used. The growth rate (µ) values were: 1.95, 0.66, 0.32, 1.89 and 1.86 h⁻¹ in MRS medium without salts and with GCA, GDCA, TCA and TDCA, respectively. Estimated doubling time was: 0.35, 0.36 and 0.37 h for MRS medium without salts, and with TCA and TDCA, respectively. For GCA and GDCA, a decrease in growth was observed from 4 h and 2 h of incubation, respectively. Bile salt hydrolase activity was determined qualitatively and quantitatively. L. casei Shirota showed activity on taurine-conjugated bile salts. The zeta potential was determined by using the same conditions as those for quantification of bile salt hydrolase activity and different potentials for each dispersion system were found; the zeta potential values were related to the metabolism of L. casei Shirota.

Keywords: Lactobacillus casei Shirota, growth rate, bile salts, morphometric parameters, bile salt hydrolase activity, zeta potential.

Se estudió el efecto de sales biliares primarias y secundarias, conjugadas a glicina[ácidoglicocólico (GCA), ácido glicodeoxicólico (GDCA)] y taurina [ácido taurocólico (TCA), ácido taurodexicólico (TDCA)] sobre los parámetros morfométricos y la tasa de crecimiento de Lactobacillus casei Shirota. Imágenes obtenidas por microscopia electrónica de barrido mostraron modificación (p < 0.05) del eje menor y la relación de aspecto celular. El primero fue modificado por la sal primaria GCA y el segundo por todas las sales. La tasa especifica de crecimiento (µ) fue: 1.95, 0.66, 0.32, 1.89 y 1.86 h⁻¹ en medio MRS sin sales, con GCA, GDCA, TCA y TDCA, respectivamente. El tiempo de duplicación estimado fue: 0.35, 0.36 y 0.37 h para medio MRS sin sales, TCA y TDCA, respectivamente. Para GCA y GDCA, el crecimiento disminuyó desde las 4 y 2 h de incubación, respectivamente. La actividad sal biliar hidrolasa fue determinada cualitativa y cuantitativamente. L. casei Shirota mostró actividad sobre sales conjugadas a taurina. Se determinó el potencial zeta para L. casei Shirota baja las mismas condiciones que para la cuantificación de la actividad sal biliar hidrolasa y se encontraron diferentes valores para cada sistema de dispersión asociados al metabolismo de la bacteria.

Palabras clave: Lactobacillus casei Shirota, tasa de crecimiento, sales biliares, parámetros morfométricos, actividad sal biliar hidrolasa, potencial zeta.

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Acknowledgements

The first author thanks the financial support given by the Mexican National Council for Science and Technology (CONACyT) in the form of a study grant (number 206844) awarded to pursue a Ph. D. in Biotechnology in the Instituto Politécnico Nacional-Escuela Nacional de Ciencias Biológicas, which is included in the record of graduate programs of excellence. Authors thank financial support by ICyTDF project 252/2010.

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