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Influence of vitamins and osmolites on growth and bacteriocin production by Lactobacillus salivarius CRL 1328 in a chemically defined medium

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Abstract:

The aim of this study was to analyze the influence of vitamins, glycerol, and salts on the growth and bacteriocin production by Lactobacillus salivarius CRL 1328, a human vagina isolate, by using a chemically defined medium to determine the optimal conditions for salivaricin production. The single omission of d-biotin, thiamine, p-aminobenzoic acid, folic acid, or cyanocobalamin did not affect the bacterial growth, whereas the removal of nicotinic acid, riboflavin, and pyridoxal produced a decrease of about 30% in the growth rate. Maximum salivaricin activity was observed after the addition of 5 or 10 g/L of NaCl. On the basis of the nutritional requirements and the levels of salivaricin production, a new optimized and simplified defined medium (SDM-NaCl) for L. salivarius CRL 1328 bacteriocin production was formulated. The kinetics of salivaricin production in SDM-NaCl and in the complex media LAPTg revealed that bacteriocin production was growth linked. A combination of tricine - sodium dodecyl sulfate polyacrylamide gel electrophoresis (Tricine-SDS-PAGE), Lumitein protein gel staining, and a bioassay for antibacterial activity indicated that the molecular mass of salivaricin CRL 1328 is about 4.5 kDa. The partially purified bacteriocin, obtained from SDM-NaCl after concentration, allowed for the design of a relatively simple method for the recovery of a biologically active protein.

Le but de cette étude était d’analyser l’influence des vitamines, du glycérol et des sels sur la croissance et la production de bactériocines par Lactobacillus salivarius CRL 1328, un isolat vaginal humain, en utilisant un milieu de culture chimique défini, afin de déterminer les conditions optimales de production de salivaricine. L’omission simple de d-biotine, de thiamine, d’acide p-aminobenzoïque, d’acide folique ou de cyanocobalamine n’affectait pas la croissance bactérienne, alors que le retrait d’acide nicotinique, de riboflavine et de pyridoxal diminuait le taux de croissance d’environ 30 %. L’activité maximale liée à la salivaricine était observée après l’ajout de 5 ou 10 g/L de NaCl. Sur la base des besoins nutritionnels et des niveaux de production de salivaricine, un nouveau milieu défini optimal et simplifié (SDM-NaCl) a été formulé pour la production de salivaricine par L. Salivarius CRL 1328. Les cinétiques de production de salivaricine dans le milieu SDM-NaCl et dans le milieu complexe LAPTg ont révélé que la production de la bactériocine était liée à la croissance. La combinaison d’une électrophorèse sur gel de polyacrylamide Tricine-SDS, d’une coloration protéique sur gel par la Lumitein et d’un essai biologique de l’activité antibactérienne a indiqué que le poids moléculaire de la salivaricine de CRL 1328 est d’environ 4,5 kDa. La bactériocine partiellement purifiée du milieu SDM-NaCl concentré a permis d’élaborer une méthode relativement simple de récupération de la protéine biologiquement active.

Document Type: Research Article

Publication date: March 1, 2009

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  • Published since 1954, this monthly journal contains new research in the field of microbiology including applied microbiology and biotechnology; microbial structure and function; fungi and other eucaryotic protists; infection and immunity; microbial ecology; physiology, metabolism and enzymology; and virology, genetics, and molecular biology. It also publishes review articles and notes on an occasional basis, contributed by recognized scientists worldwide.
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