Lactate efflux-induced electrical potential in membrane vesicles of Streptococcus cremoris

We developed a procedure for isolating membrane vesicles from the homolactic fermentative bacterium Streptococcus cremoris. The membrane vesicles were shown to have a right-side-out orientation by freeze-etch electron microscopy and to be free of cytoplasmic constituents. The membrane vesicles retained their functional properties and accumulated the amino acids L-leucine, L-histidine, and L-alanine in response to a valinomycin-induced potassium diffusion gradient. Studies with these membrane vesicles strongly supported the possibility that there was a proton motive force-generating mechanism by end product efflux (Michels et al., FEMS Lett. 5:357-364, 1979). Lactate efflux from membrane vesicles which were loaded with L-lactate and diluted in a lactate-free medium led to the generation of an electrical potential across the membrane. The results indicate that lactate efflux is an electrogenic process by which L-lactate is translocated with more than one proton.