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.

doi:10.3390/microorganisms11020294

Open AccessArticle

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

by

Evgeniya A. Malykh

¹,

Liubov I. Golubeva

¹,

Ekaterina S. Kovaleva

¹,

Mikhail S. Shupletsov

^1,2,

Elena V. Rodina

³,

Sergey V. Mashko

^1,4 and

Nataliya V. Stoynova

^1,*

¹

Ajinomoto-Genetika Research Institute, 117545 Moscow, Russia

²

Computational Mathematics and Cybernetics Department, Lomonosov Moscow State University, 119991 Moscow, Russia

³

Chemistry Department, Lomonosov Moscow State University, 119991 Moscow, Russia

⁴

Biological Department, Lomonosov Moscow State University, 119991 Moscow, Russia

^*

Author to whom correspondence should be addressed.

Microorganisms 2023, 11(2), 294; https://doi.org/10.3390/microorganisms11020294

Submission received: 18 November 2022 / Revised: 11 January 2023 / Accepted: 14 January 2023 / Published: 23 January 2023

(This article belongs to the Special Issue Microbial Cell Factories: Production of Amino Acids Using Microorganisms)

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Abstract

Inorganic pyrophosphatases (PPases) catalyze an essential reaction, namely, the hydrolysis of PP_i, which is formed in large quantities as a side product of numerous cellular reactions. In the majority of living species, PP_i 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⁺-PPase^Rru) in the form of a proton electrochemical gradient for further ATP synthesis. Here, the codon-harmonized gene hppa^Rru encoding H⁺-PPase^Rru was expressed in the Escherichia coli chromosome. We demonstrate, for the first time, that H⁺-PPase^Rru complements the essential native S-PPase in E. coli cells. ¹³C-MFA confirmed that replacing native PPase to H⁺-PPase^Rru 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.

Keywords: inorganic pyrophosphatase; Rhodospirillum rubrum; ¹³C-MFA; H⁺-translocating pyrophosphatase; soluble pyrophosphatase; metabolic engineering; pyrophosphate hydrolysis; ATP; synthetic biology

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MDPI and ACS Style

Malykh, E.A.; Golubeva, L.I.; Kovaleva, E.S.; Shupletsov, M.S.; Rodina, E.V.; Mashko, S.V.; Stoynova, N.V. H⁺-Translocating Membrane-Bound Pyrophosphatase from Rhodospirillum rubrum Fuels Escherichia coli Cells via an Alternative Pathway for Energy Generation. Microorganisms 2023, 11, 294. https://doi.org/10.3390/microorganisms11020294

AMA Style

Malykh EA, Golubeva LI, Kovaleva ES, Shupletsov MS, Rodina EV, Mashko SV, Stoynova NV. H⁺-Translocating Membrane-Bound Pyrophosphatase from Rhodospirillum rubrum Fuels Escherichia coli Cells via an Alternative Pathway for Energy Generation. Microorganisms. 2023; 11(2):294. https://doi.org/10.3390/microorganisms11020294

Chicago/Turabian Style

Malykh, Evgeniya A., Liubov I. Golubeva, Ekaterina S. Kovaleva, Mikhail S. Shupletsov, Elena V. Rodina, Sergey V. Mashko, and Nataliya V. Stoynova. 2023. "H⁺-Translocating Membrane-Bound Pyrophosphatase from Rhodospirillum rubrum Fuels Escherichia coli Cells via an Alternative Pathway for Energy Generation" Microorganisms 11, no. 2: 294. https://doi.org/10.3390/microorganisms11020294

APA Style

Malykh, E. A., Golubeva, L. I., Kovaleva, E. S., Shupletsov, M. S., Rodina, E. V., Mashko, S. V., & Stoynova, N. V. (2023). H⁺-Translocating Membrane-Bound Pyrophosphatase from Rhodospirillum rubrum Fuels Escherichia coli Cells via an Alternative Pathway for Energy Generation. Microorganisms, 11(2), 294. https://doi.org/10.3390/microorganisms11020294

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

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