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H+-Translocating Membrane-Bound Pyrophosphatase from Rhodospirillum rubrum Fuels Escherichia coli Cells via an Alternative Pathway for Energy Generation.
Malykh, Evgeniya A; Golubeva, Liubov I; Kovaleva, Ekaterina S; Shupletsov, Mikhail S; Rodina, Elena V; Mashko, Sergey V; Stoynova, Nataliya V.
Afiliación
  • Malykh EA; Ajinomoto-Genetika Research Institute, 117545 Moscow, Russia.
  • Golubeva LI; Ajinomoto-Genetika Research Institute, 117545 Moscow, Russia.
  • Kovaleva ES; Ajinomoto-Genetika Research Institute, 117545 Moscow, Russia.
  • Shupletsov MS; Ajinomoto-Genetika Research Institute, 117545 Moscow, Russia.
  • Rodina EV; Computational Mathematics and Cybernetics Department, Lomonosov Moscow State University, 119991 Moscow, Russia.
  • Mashko SV; Chemistry Department, Lomonosov Moscow State University, 119991 Moscow, Russia.
  • Stoynova NV; Ajinomoto-Genetika Research Institute, 117545 Moscow, Russia.
Microorganisms ; 11(2)2023 Jan 23.
Article en En | MEDLINE | ID: mdl-36838259
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.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Idioma: En Revista: Microorganisms Año: 2023 Tipo del documento: Article País de afiliación: Rusia Pais de publicación: Suiza

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Idioma: En Revista: Microorganisms Año: 2023 Tipo del documento: Article País de afiliación: Rusia Pais de publicación: Suiza