Journal of Bacteriology · 2007 · 73 citations · 48 references
Life SciencesBiologyMicrobiologyBioenergeticsMembrane BiophysicsMicrobial MetabolismCellular EnergeticsElectron Transport ChainLactic Acid BacteriaMembrane BioenergeticsMembrane PotentialProton Motive ForceBacterial Growth YieldBacterial Proton Motive ForceFluorescent ProbesLactic Acid Bacteria PhysiologyMembrane Potential MeasurementNadh Dehydrogenase ActivityOxygen Consumption KineticsRespiratory Chain Reactions
Lactococcus lactis, a facultative anaerobic lactic acid bacterium, is known to have an increased growth yield when grown aerobically in the presence of heme. We have now established the presence of a functional, proton motive force-generating electron transfer chain (ETC) in L. lactis under these conditions. Proton motive force generation in whole cells was measured using a fluorescent probe (3',3'-dipropylthiadicarbocyanine), which is sensitive to changes in membrane potential (Delta psi). Wild-type cells, grown aerobically in the presence of heme, generated a Delta psi even in the presence of the F(1)-F(o) ATPase inhibitor N,N'-dicyclohexylcarbodiimide, while a cytochrome bd-negative mutant strain (CydA Delta) did not. We also observed high oxygen consumption rates by membrane vesicles prepared from heme-grown cells, compared to CydA Delta cells, upon the addition of NADH. This demonstrates that NADH is an electron donor for the L. lactis ETC and demonstrates the presence of a membrane-bound NADH-dehydrogenase. Furthermore, we show that the functional respiratory chain is present throughout the exponential and late phases of growth.
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Molecular Cloning. A Laboratory Manual
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Measurement of protein using bicinchoninic acid
Pam Smith, Randall I. Krohn, Greg T. Hermanson et al. · Analytical Biochemistry · 1985 · 18.4K citations