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Journal of Bacteriology, June 2001, p. 3458-3467, Vol. 183, No. 11
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.11.3458-3467.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Twofold Reduction of Phosphofructokinase Activity in Lactococcus lactis Results in Strong Decreases in Growth Rate and in Glycolytic Flux

Heidi W. Andersen, Christian Solem, Karin Hammer, and Peter R. Jensen*

Section of Molecular Microbiology, BioCentrum, Technical University of Denmark, DK-2800 Lyngby, Denmark

Received 16 January 2001/Accepted 21 March 2001

Two mutant strains of Lactococcus lactis in which the promoter of the las operon, harboring pfk, pyk, and ldh, were replaced by synthetic promoters were constructed. These las mutants had an approximately twofold decrease in the activity of phosphofructokinase, whereas the activities of pyruvate kinase and lactate dehydrogenase remained closer to the wild-type level. In defined medium supplemented with glucose, the growth rate of the mutants was reduced to 57 to 70% of wild-type levels and the glycolytic flux was reduced to 62 to 76% of wild-type levels. In complex medium growth was even further reduced. Surprisingly, the mutants still showed homolactic fermentation, which indicated that the limitation was different from standard glucose-limited conditions. One explanation could be that the reduced activity of phosphofructokinase resulted in the accumulation of sugar-phosphates. Indeed, when one of the mutants was starved for glucose in glucose-limited chemostat, the growth rate could gradually be increased to 195% of the growth rate observed in glucose-saturated batch culture, suggesting that phosphofructokinase does affect the concentration of upstream metabolites. The pools of glucose-6-phosphate and fructose-6-phosphate were subsequently found to be increased two- to fourfold in the las mutants, which indicates that phosphofructokinase exerts strong control over the concentration of these metabolites.

* Corresponding author. Mailing address: Section of Molecular Microbiology, BioCentrum, Technical University of Denmark, Building 301, DK-2800 Lyngby, Denmark. Phone: 45 45-252510. Fax: 45 45-932809. E-mail: Peter.R.Jensen{at}BioCentrum.DTU.DK.

Journal of Bacteriology, June 2001, p. 3458-3467, Vol. 183, No. 11
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.11.3458-3467.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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