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Analysis of ldh genes in Lactobacillus casei BL23: role on lactic acid production

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Journal of Industrial Microbiology & Biotechnology

Abstract

Lactobacillus casei is a lactic acid bacterium that produces l-lactate as the main product of sugar fermentation via l-lactate dehydrogenase (Ldh1) activity. In addition, small amounts of the d-lactate isomer are produced by the activity of a d-hydroxycaproate dehydrogenase (HicD). Ldh1 is the main l-lactate producing enzyme, but mutation of its gene does not eliminate l-lactate synthesis. A survey of the L. casei BL23 draft genome sequence revealed the presence of three additional genes encoding Ldh paralogs. In order to study the contribution of these genes to the global lactate production in this organism, individual, as well as double mutants (ldh1 ldh2, ldh1 ldh3, ldh1 ldh4 and ldh1 hicD) were constructed and lactic acid production was assessed in culture supernatants. ldh2, ldh3 and ldh4 genes play a minor role in lactate production, as their single mutation or a mutation in combination with an ldh1 deletion had a low impact on l-lactate synthesis. A Δldh1 mutant displayed an increased production of d-lactate, which was probably synthesized via the activity of HicD, as it was abolished in a Δldh1 hicD double mutant. Contrarily to HicD, no Ldh1, Ldh2, Ldh3 or Ldh4 activities could be detected by zymogram assays. In addition, these assays revealed the presence of extra bands exhibiting d-/l-lactate dehydrogenase activity, which could not be attributed to any of the described genes. These results suggest that L. casei BL23 possesses a complex enzymatic system able to reduce pyruvic to lactic acid.

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Acknowledgments

J. Rico was recipient of a CSIC postgraduate fellowship partially funded by the Corporación Alimentaria Peñasanta. Genome sequencing of L. casei BL23 was carried out at the University of Caen, Laboratoire de Microbiologie de l’Environnement, at the INRA Thiverval-Grignon, Microbiologie et Génétique Moléculaire and at the Instituto de Agroquímica y Tecnología de Alimentos, CSIC, with the financial aide from the Region Basse Normandie and the INRA.

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Authors and Affiliations

  1. Laboratorio de Bacterias Lácticas y Probióticos, IATA-CSIC, P.O. Box 73, 46100, Burjassot, Valencia, Spain

    Juan Rico, María Jesús Yebra, Gaspar Pérez-Martínez & Vicente Monedero

  2. Laboratorie de Microbiologie et Génétique Moleculaire, AgroParisTech-CNRS-INRA, 78850, Thiverval-Grignon, France

    Josef Deutscher

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  1. Juan Rico
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  2. María Jesús Yebra
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  5. Vicente Monedero
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Correspondence to Vicente Monedero.

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Rico, J., Yebra, M.J., Pérez-Martínez, G. et al. Analysis of ldh genes in Lactobacillus casei BL23: role on lactic acid production. J Ind Microbiol Biotechnol 35, 579–586 (2008). https://doi.org/10.1007/s10295-008-0319-8

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