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The relation of proton motive force, adenylate energy charge and phosphorylation potential to the specific growth rate and efficiency of energy transduction inBacillus licheniformis under aerobic growth conditions

Abstract

The magnitude of the proton motive force (Δp) and its constituents, the electrical (Δψ) and chemical potential (-ZΔpH), were established for chemostat cultures of a protease-producing, relaxed (rel ) variant and a not protease-producing, stringent (rel +) variant of an industrial strain ofBacillus licheniformis (respectively referred to as the A- and the B-type). For both types, an inverse relation of Δp with the specific growth rate μ was found. The calculated intracellular pH (pHin) was not constant but inversely related to μ. This change in pHin might be related to regulatory functions of metabolism but a regulatory role for pHin itself could not be envisaged. Measurement of the adenylate energy charge (EC) showed a direct relation with μ for glucose-limited chemostat cultures; in nitrogen-limited chemostat cultures, the EC showed an approximately constant value at low μ and an increased value at higher μ. For both limitations, the ATP/ADP ratio was directly related to μ.

The phosphorylation potential (ΔG'p) was invariant with μ. From the values for ΔG'p and Δp, a variable →H+/ATP-stoichiometry was inferred: →H+/ATP=1.83+0.52µ, so that at a given →H+/O-ratio of four (4), the apparent P/O-ratio (inferred from regression analysis) showed a decline of 2.16 to 1.87 for μ=0 to μmax (we discuss how more than half of this decline will be independent of any change in internal cell-volume). We propose that the constancy of ΔG'p and the decrease in the efficiency of energy-conservation (P/O-value) with increasing μ are a way in which the cells try to cope with an apparent less than perfect coordination between anabolism and catabolism to keep up the highest possible μ with a minimum loss of growth-efficiency. Protease production in nitrogen-limited cultures as compared to glucose-limited cultures, and the difference between the A- and B-type, could not be explained by a different energy-status of the cells.

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Abbreviations

CCCP:

carbonylcyanide-p-trichloromethoxyphenylhydrazone

DW:

dry weight of biomass

F:

Faraday's constant, 96.6 J/(mV × mol)

Fo:

chemostat outflow-rate (ml/h)

FCCP:

carbonylcyanide-p-trifluoromethoxyphenylhydrazone

ΔG'p:

phosphorylation potential, the Gibbs energy change for ATP-synthesis from ADP and Pi

ΔG'0p:

standard Gibbs energy change at specified conditions

→H+/ATP:

number of protons translocated through

ATP:

synthase in synthesis of one ATP

→H+/O:

protons translocated during transfer of 2 electrons from substrate to oxygen

μ:

specific growth rate (1/h)

ΔμH+:

transmembrane electrochemical proton potential, J/mol

Mb :

‘molar weight’ (147.6 g/mol) of bacteria with general cell formula C6.0H10.8O3.0N1.2

pHout,in :

extracellular, intracellular pH

Pi :

(intracellular) inorganic phosphate

Δp:

proton motive force, mV

ΔpH:

transmembrane pH-difference

Δψ:

transmembrane electrical potential, mV

P/O:

number of ADP phosphorylated to ATP upon reduction of one ‘O2−’ to H2O by two electrons transferred through the electron transfer chain

P/O:

(→H+/O) × (→H+/ATP)−1

P/OF, P/ON :

P/O with the two electrons donated by resp. (NADH + H+) and FADH

q:

specific rate of consumption or production (mol/g DW × h)

rel +,rel :

stringent, relaxed genotype

R:

universal gas constant, 8.36 J/(mol × degree)

T:

absolute temperature

TPMP+ :

triphenylmethylphosphonium ion

TPP+ :

tetraphenyl phosphonium ion

Y:

growth yield, g DW/mol

Z:

conversion constant=61.8 mV for 310 K (37 °C)

ZΔpH:

transmembrane proton potential or chemical potential, mV

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Correspondence to Henk W. van Verseveld.

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Bulthuis, B.A., Koningstein, G.M., Stouthamer, A.H. et al. The relation of proton motive force, adenylate energy charge and phosphorylation potential to the specific growth rate and efficiency of energy transduction inBacillus licheniformis under aerobic growth conditions. Antonie van Leeuwenhoek 63, 1–16 (1993). https://doi.org/10.1007/BF00871725

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Key words

  • adenylate energy charge
  • Bacillus
  • chemostat
  • energy conservation
  • extracellular protease
  • membrane potential
  • phosphorylation potential
  • proton motive force