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H+-Translocating Membrane-Bound Pyrophosphatase from Rhodospirillum rubrum Fuels Escherichia coli Cells via an Alternative Pathway for Energy Generation.


ABSTRACT: Inorganic pyrophosphatases (PPases) catalyze an essential reaction, namely, the hydrolysis of PPi, which is formed in large quantities as a side product of numerous cellular reactions. In the majority of living species, PPi hydrolysis is carried out by soluble cytoplasmic PPase (S-PPases) with the released energy dissipated in the form of heat. In Rhodospirillum rubrum, part of this energy can be conserved by proton-pumping pyrophosphatase (H+-PPaseRru) in the form of a proton electrochemical gradient for further ATP synthesis. Here, the codon-harmonized gene hppaRru encoding H+-PPaseRru was expressed in the Escherichia coli chromosome. We demonstrate, for the first time, that H+-PPaseRru complements the essential native S-PPase in E. coli cells. 13C-MFA confirmed that replacing native PPase to H+-PPaseRru leads to the re-distribution of carbon fluxes; a statistically significant 36% decrease in tricarboxylic acid (TCA) cycle fluxes was found compared with wild-type E. coli MG1655. Such a flux re-distribution can indicate the presence of an additional method for energy generation (e.g., ATP), which can be useful for the microbiological production of a number of compounds, the biosynthesis of which requires the consumption of ATP.

SUBMITTER: Malykh EA 

PROVIDER: S-EPMC9959109 | biostudies-literature | 2023 Jan

REPOSITORIES: biostudies-literature

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H<sup>+</sup>-Translocating Membrane-Bound Pyrophosphatase from <i>Rhodospirillum rubrum</i> Fuels <i>Escherichia coli</i> Cells via an Alternative Pathway for Energy Generation.

Malykh Evgeniya A EA   Golubeva Liubov I LI   Kovaleva Ekaterina S ES   Shupletsov Mikhail S MS   Rodina Elena V EV   Mashko Sergey V SV   Stoynova Nataliya V NV  

Microorganisms 20230123 2


Inorganic pyrophosphatases (PPases) catalyze an essential reaction, namely, the hydrolysis of PP<sub>i</sub>, which is formed in large quantities as a side product of numerous cellular reactions. In the majority of living species, PP<sub>i</sub> hydrolysis is carried out by soluble cytoplasmic PPase (S-PPases) with the released energy dissipated in the form of heat. In <i>Rhodospirillum rubrum,</i> part of this energy can be conserved by proton-pumping pyrophosphatase (H<sup>+</sup>-PPase<sup>Rr  ...[more]

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