Abstract
Respiratory chain phosphorylation has been investigated in the methylotrophic bacterium Methylophilus methylotrophus following the addition of oxidisable substrates to aerobic, whole cell suspensions. Initial-rate experiments showed that ATP synthesis occurred at the overall expense of AMP and inorganic phosphate via the sequential action of the ATP phosphohydrolase and adenylate kinase; some of the nascent ATP was rapidly used to synthesis nonadenine nucleoside triphosphates. After being corrected for ATP turnover, Pi/O quotients of 0.46 to 0.54, 0.77 and 1.37 nmol/ng-atom O were obtained for the oxidation of methanol dehydrogenase-linked substrates (methanol, ethanol and acetaldehyde), duroquinol and formate (NAD+-linked) respectively. These values were proportional to the →H+/O and/or →K+/O quotients exhibited by these substrates, and yielded an average →H+/ATP (≡→H+/Pi) quotient of 4.2 ng-ion H+/nmol. Steady-state experiments showed that the extent of cellular energisation varied with the respiration rate but was always in the order methanol > duroquinol > acetaldehyde, thus indicating that under these longer-term conditions methanol was completely oxidised to yield PQQH2 and 2NAD(P)H. These results are discussed in terms of the various reactions which lead to the generation or utilisation of the protonmotive force in this organism.
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Abbreviations
- FCCP:
-
carbonylcyanide p-trifluoromethyxyphenyl-hydrazone
- \(\Delta \bar \mu {\text{H}}^ +\) :
-
bulk phase, transmembrane electrochemical potential difference of protons (\((\bar \mu \Psi ){\text{H}}_{{\text{in}}}^{\text{ + }} - \bar \mu {\text{H}}_{{\text{out}}}^{\text{ + }} )\))
- ΔpH:
-
bulk phase, transmembrane pH difference (pHin−pHout)
- ΔΨ:
-
bulk phase, transmembrane electrical potential difference (ψin - ψout)
- [∼P]:
-
concentration of anhydride phosphate bonds in adenine nucleotides (2[ATP]+[ADP])
- FPLC:
-
fast protein liquid chromatography
- PQQ:
-
pyrroloquinoline quinone
- ΔGp:
-
phosphorylation potential
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Patchett, R.A., Quilter, J.A. & Jones, C.W. Energy conservation in whole cells of the methylotrophic bacterium Methylophilus methylotrophus . Arch. Microbiol. 141, 95–102 (1985). https://doi.org/10.1007/BF00446747
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DOI: https://doi.org/10.1007/BF00446747