Twofold reduction of phosphofructokinase activity in Lactococcus lactis results in strong decreases in growth rate and in glycolytic flux

Journal of Bacteriology
H W AndersenP R Jensen

Abstract

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 wer...Continue Reading

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Citations

May 17, 2011·Microbial Ecology·Tjaša Danevčič, David Stopar
Jan 7, 2004·Journal of Microbiological Methods·Dina Petranovic, Ivan Mijakovic
Feb 6, 2004·Trends in Biotechnology·Willem M de Vos, Jeroen Hugenholtz
Apr 27, 2002·Applied and Environmental Microbiology·Christian Solem, Peter Ruhdal Jensen
Aug 30, 2002·Applied and Environmental Microbiology·Brian J KoebmannPeter R Jensen
Feb 7, 2003·Applied and Environmental Microbiology·Ingeborg C BoelsWillem M de Vos
Mar 5, 2005·Applied and Environmental Microbiology·H Wouter WisselinkJeroen Hugenholtz
Jul 10, 2007·Journal of Bacteriology·Christian SolemPeter R Jensen
Jun 26, 2002·Journal of Bacteriology·Brian J KoebmannPeter R Jensen
Nov 5, 2003·Journal of Bacteriology·Casper Møller JørgensenJan Martinussen
Nov 1, 2011·PloS One·Sandra M CarvalhoAna Rute Neves
Mar 19, 2014·PloS One·Siu Hung Joshua ChanPeter Ruhdal Jensen
Oct 27, 2015·Frontiers in Microbiology·Laura PaixãoAna R Neves
Apr 28, 2005·The FEBS Journal·Brian KoebmannPeter Ruhdal Jensen
Jan 1, 2012·Computational and Structural Biotechnology Journal·Maria Papagianni
Jun 4, 2011·International Journal of Food Microbiology·Irene BustosM Carmen Martínez-Cuesta
Apr 25, 2007·Critical Reviews in Microbiology·Magdalena Kowalczyk, Jacek Bardowski
Oct 26, 2014·Applied and Environmental Microbiology·Onur ErcanMichiel Kleerebezem
Sep 1, 2018·FEMS Microbiology Letters·Rajni Hatti-KaulHesham El Enshasy
May 18, 2010·Journal of Basic Microbiology·María-Enriqueta Muñoz-Márquez, Elizabeth Ponce-Rivas

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