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Antonie van Leeuwenhoek

, Volume 63, Issue 1, pp 1–16 | Cite as

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

  • Ben A. Bulthuis
  • Gregory M. Koningstein
  • Adriaan H. Stouthamer
  • Henk W. van Verseveld
Article

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.

Key words

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

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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Copyright information

© Kluwer Academic Publishers 1993

Authors and Affiliations

  • Ben A. Bulthuis
    • 1
  • Gregory M. Koningstein
    • 1
  • Adriaan H. Stouthamer
    • 1
  • Henk W. van Verseveld
    • 1
  1. 1.Department of Microbiology, Biological LaboratoryVrije UniversiteitAmsterdamThe Netherlands

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