An alkaline-tolerant bacterium, which grew on various carbohydrates between pH 6 and 9·5, was isolated from soil and identified as Bacillus circulans. Lactose-grown organisms exhibited a transmembrane pH gradient (ΔpH) of −47 mV at pH 6·6, but no ΔpH at pH 9·0. The transmembrane electrical potential (Δψ) was −66 mV at pH 6·6 and −115mV at pH 9·0. Thus the total protonmotive forces at the two pH values were essentially the same. Lactose-grown organisms transported thiomethyl β-D-galactopyranoside (TMG) at pH 6·6 and at pH 9·0, but transport at the alkaline pH was dependent upon addition of ascorbate/N,N,N′,N′-tetramethyl-p-phenylenediamine (TMPD) or preincubation with lactose. In the presence of ascorbate/TMPD, the TMG transport system exhibited similar kinetics and substrate specificities at pH 6·6 and pH 9·0, and resulted in accumulation of chemically unmodified TMG to a concentration approximately 180 times greater than the external concentration. Experiments in which a diffusion potential was generated in starved organisms or in which organisms were treated with nigericin indicated a lack of correlation between the rate of TMG uptake and the magnitude of Δψ. By contrast, the rate of TMG uptake correlated with cellular ATP levels in organisms incubated at different pH values and in organisms treated with N,N′-dicyclohexylcarbodiimide, arsenate or nigericin.
BergerE. A.1973; Different mechanisms of energy coupling for the active transport of proline and glutamine in Escherichia coli. Proceedings of the National Academy of Sciences of the United States of America 70:1514–1518
BergerE. A.,
HeppelL.A.1974; Different mechanisms of energy coupling for the shock-sensitive and shock-resistant amino acid permeases of Escherichia coli. Journal of Biological Chemistry 249:7747–7755
BisschopA.,
de JongL.,
Lima CostaM. E.,
KoningsW. N.1975a; Relation between reduced nicotinamide adenine dinucleotide oxidation and amino acid transport in membrane vesicles from Bacillus subtilis. Journal of Bacteriology 121:807–813
ChislettM. E.,
KushnerD. J.1961; A strain of Bacillus circulans capable of growing under highly alkaline conditions. Journal of General Microbiology 24:187–190
ClarkV. L.,
YoungF. E.1974; Active transport of D-alanine and related amino acids by whole cells of Bacillus subtilis. Journal of Bacteriology 120:1085–1092
ColeH.,
WimpennyJ. W. T.,
HughesD. E.1967; The ATP pool in Escherichia coli I measurement of the pool using a modified luciferase assay. Biochimica et biophysica acta 143:445–453
FlaggJ. L.,
WilsonT. H.1977; A proton-motive force as the source of energy for galactoside transport in energy-depleted Escherichia coli. Journal of Membrane Biology 31:233–255
GuffantiA. A.,
SusmanP.,
BlancoR.,
Krul-WichT. A.1978; The protonmotive force and α-aminoisobutyric acid transport in an obligately alkalophilic bacterium. Journal of Biological Chemistry 253:708–715
HegemanG.D.1966; Synthesis of the enzymes of the mandelate pathway by Pseudomonas putida I Synthesis of enzymes by the wild type. Journal of Bacteriology 91:1140–1154
HertzbergE. L.,
HinkleP. C.1974; Oxidative phosphorylation and proton translocation in membrane vesicles prepared from Escherichia coli. Biochemical and Biophysical Research Communications 58:178–184
KashketE. R.,
WilsonT. H.1974; Proton-motive force in fermenting Streptococcus lactis 7962 in relation to sugar accumulation. Biochemical and Biophysical Research Communications 59:879–885
KennedyE. P.1970; The lactose permease system of Escherichia coli. The Lactose Operon49–92 Edited by
BeckwithJ. R.,
ZipserD.
New York: Cold Spring Harbor Laboratory;
KoningsW. N.,
BarnesE. M.,
KabackH. R.1971; Mechanisms of active transport in isolated membrane vesicles III. The coupling of reduced phenazine methosulfate to the concentrative uptake of β-galactosides and amino acids. Journal of Biological Chemistry 246:5857–5861
KushnerD. J.,
LissonT. A.1959; Alkali resistance in a strain of Bacillus cereus pathogenic for the Larch Sawfly Pristiphora erichsonii. Journal of General Microbiology 21:96–108
LarsenS. H.,
AdlerJ.,
GargusJ. J.,
HoggR. W.1974; Chemomechanical coupling without ATP: the source of energy for motility and chemotaxis in bacteria. Proceedings of the National Academy of Sciences of the United States of America 71:1239–1243
LowryO. H.,
RosebroughN. J.,
FarrA. L/,
RandallR. J.1951; Protein measurement with the Folin phenol reagent. Journal of Biological Chemistry 193:265–275
MaloneyP. C.,
KashketE. R.,
WilsonT. H.1974; A proton motive force drives ATP synthesis in bacteria. Proceedings of the National Academy of Sciences of the United States of America 71:3896–3900
MaloneyP. C.,
KashketE. R.,
WilsonT. H.1975 Methods for studying transport in bacteria. In Methods in Membrane Biology31–49 Edited by
KornE. D.
New York: Plenum Press;
MarmurJ.,
DotyP.1959; Heterogeneity in deoxyribonucleic acids I. Dependence on composition of the configurational stability of deoxyribonucleic acids. Nature, London 183:1427–1429
PavlasovaE.,
HaroldF. M.1968; Energy coupling in the transport of β-galactosides by Escherichia coli: effect of proton conductors. Journal of Bacteriology 98:198–204
RamosS.,
KabackH. R.1977b; The relationship between the electrochemical proton gradient and active transport in Escherichia coli membrane vesicles. Biochemistry 16:854–859
RamosS.,
SchuldinerS.,
KabackH. R.1976; The electrical gradient of protons and its relationship to active transport in Escherichia coli membrane vesicles. Proceedings of the National Academy of Sciences of the United States of America 73:1892–1896
StanleyP. E.,
WilliamsS. G.1969; Use of the liquid scintillation spectrometer for determining adenosine triphosphate by the luciferase enzyme. Analytical Biochemistry 29:381–392
TanakaS.,
LernerS. A.,
LINE. C. C.1967; Replacement of a phosphoenolpyruvate-dependent phosphotransferase by a nicotinamide adenine dinucleotide-linked dehydrogenase for the utilization of mannitol. Journal of Bacteriology 93:642–648
ThipoyathasanaP.,
ValentineR. C.1974; The requirement for energy transducing ATPase for anaerobic motility in Escherichia coli. Bio-chimica et biophysica acta 347:464–468
TsuchiyaT.1977; Adenosine 5′-triphosphate synthesis driven by a protonmotive force in membrane vesicles of Escherichia coli. Journal of Bacteriology 129:763–769
TsuchiyaT.,
RosenP.1976; Adenosine 5′-triphosphate synthesis energized by an artificially imposed membrane potential in membrane vesicles of Escherichia coli. Journal of Bacteriology 127:154–161
WaddellW. J.,
ButlerT. C.1959; Calculation of intracellular pH from the distribution of 5,5-dimethyl-2,4-oxazolidinedione (DMO): application to skeletal muscle of the dog. Journal of Clinical Investigation 38:720–729
WilsonD. M.,
AldereteJ. F.,
MaloneyP. C.,
WilsonT. H.1976; Protonmotive force as the source of energy for adenosine 5′-triphosphate synthesis in Escherichia coli. Journal of Bacteriology 126:327–337
WinklerH. H.,
WilsonT. H.1966; The role of energy coupling in the transport of β-galacto-sides by Escherichia coli. Journal of Biological Chemistry 241:2200–2211