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Journal of Bacteriology, July 2001, p. 3817-3824, Vol. 183, No. 13
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.13.3817-3824.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Molecular Physiology of Sugar Catabolism in Lactococcus lactis IL1403

Sergine Even, Nic D. Lindley, and Muriel Cocaign-Bousquet*

Centre de Bioingénierie Gilbert Durand, UMR 5504 INSA/CNRS and UMR 792 INSA/INRA, Institut National des Sciences Appliquées, 31077 Toulouse Cedex 4, France

Received 8 January 2001/Accepted 5 April 2001

The metabolic characteristics of Lactococcus lactis IL1403 were examined on two different growth media with respect to the physiological response to two sugars, glucose and galactose. Analysis of specific metabolic rates indicated that despite significant variations in the rates of both growth and sugar consumption, homolactic fermentation was maintained for all cultures due to the low concentration of either pyruvate-formate lyase or alcohol dehydrogenase. When the ionophore monensin was added to the medium, flux through glycolysis was not increased, suggesting a catabolic flux limitation, which, with the low intracellular concentrations of glycolytic intermediates and high in vivo glycolytic enzyme capacities, may be at the level of sugar transport. To assess transcription, a novel DNA macroarray technology employed RNA labeled in vitro with digoxigenin and detection of hybrids with an alkaline phosphatase-antidigoxigenin conjugate. This method showed that several genes of glycolysis were expressed to higher levels on glucose and that the genes of the mixed-acid pathway were expressed to higher levels on galactose. When rates of enzyme synthesis are compared to transcript concentrations, it can be deduced that some translational regulation occurs with threefold-higher translational efficiency in cells grown on glucose.

* Corresponding author. Mailing address: Centre de Bioingénierie Gilbert Durand, UMR 5504 INSA/CNRS & UMR 792 INSA/INRA, Institut National des Sciences Appliquées, 135 Ave. de Rangueil, 31077 Toulouse Cedex 4, France. Phone: (33) 561 559 438. Fax: (33) 561 559 400. E-mail: cocaign{at}insa-tlse.fr.

Journal of Bacteriology, July 2001, p. 3817-3824, Vol. 183, No. 13
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.13.3817-3824.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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