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Journal of Bacteriology, July 2007, p. 5203-5209, Vol. 189, No. 14
0021-9193/07/$08.00+0     doi:10.1128/JB.00361-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Generation of a Membrane Potential by Lactococcus lactis through Aerobic Electron Transport{triangledown}

R. J. W. Brooijmans,1 B. Poolman,2 G. K. Schuurman-Wolters,2 W. M. de Vos,1,4 and J. Hugenholtz1,3*

Kluyver Centre for Genomics of Industrial Fermentation, Wageningen Centre for Food Sciences, Wageningen,1 Membrane Enzymology Group, Department of Biochemistry, University of Groningen, Groningen,2 NIZO Food Research, Ede,3 Department of General Microbiology, Wageningen University and Research Centre, Wageningen, The Netherlands4

Received 10 March 2007/ Accepted 1 May 2007

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 F1-Fo 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.

* Corresponding author. Mailing address: NIZO Food Research, P.O. Box 20, 6710 BA Ede, The Netherlands. Phone: 31-318-659540. Fax: 31-318-650400. E-mail: jeroen.hugenholtz{at}nizo.nl

{triangledown} Published ahead of print on 11 May 2007.

Journal of Bacteriology, July 2007, p. 5203-5209, Vol. 189, No. 14
0021-9193/07/$08.00+0     doi:10.1128/JB.00361-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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