Pyrophosphate-dependent ATP formation from acetyl coenzyme a in Syntrophus aciditrophicus, a new twist on ATP formation

Kimberly L. James, Luis A. Ríos-Hernández, Neil Q. Wofford, Housna Mouttaki, Jessica R. Sieber, Cody S. Sheik, Hong H. Nguyen, Yanan Yang, Yongming Xie, Jonathan Erde, Lars Rohlin, Elizabeth A. Karr, Joseph A. Loo, Rachel R.Ogorzalek Loo, Gregory B. Hurst, Robert P. Gunsalus, Luke I. Szweda, Michael J. McInerney

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Syntrophus aciditrophicus is a model syntrophic bacterium that degrades key intermediates in anaerobic decomposition, such as benzoate, cyclohexane-1-carboxylate, and certain fatty acids, to acetate when grown with hydrogen-/formateconsuming microorganisms. ATP formation coupled to acetate production is the main source for energy conservation by S. aciditrophicus. However, the absence of homologs for phosphate acetyltransferase and acetate kinase in the genome of S. aciditrophicus leaves it unclear as to how ATP is formed, as most fermentative bacteria rely on these two enzymes to synthesize ATP from acetyl coenzyme A (CoA) and phosphate. Here, we combine transcriptomic, proteomic, metabolite, and enzymatic approaches to show that S. aciditrophicus uses AMP-forming, acetyl-CoA synthetase (Acs1) for ATP synthesis from acetyl- CoA. acs1 mRNA and Acs1 were abundant in transcriptomes and proteomes, respectively, of S. aciditrophicus grown in pure culture and coculture. Cell extracts of S. aciditrophicus had low or undetectable acetate kinase and phosphate acetyltransferase activities but had high acetyl-CoA synthetase activity under all growth conditions tested. Both Acs1 purified from S. aciditrophicus and recombinantly produced Acs1 catalyzed ATP and acetate formation from acetyl-CoA, AMP, and pyrophosphate. High pyrophosphate levels and a high AMP-to-ATP ratio (5.9 ± 1.4) in S. aciditrophicus cells support the operation of Acs1 in the acetate-forming direction. Thus, S. aciditrophicus has a unique approach to conserve energy involving pyrophosphate, AMP, acetyl-CoA, and an AMP-forming, acetyl-CoA synthetase. IMPORTANCE Bacteria use two enzymes, phosphate acetyltransferase and acetate kinase, to make ATP from acetyl-CoA, while acetate-forming archaea use a single enzyme, an ADP-forming, acetyl-CoA synthetase, to synthesize ATP and acetate from acetyl-CoA. Syntrophus aciditrophicus apparently relies on a different approach to conserve energy during acetyl-CoA metabolism, as its genome does not have homologs to the genes for phosphate acetyltransferase and acetate kinase. Here, we show that S. aciditrophicus uses an alternative approach, an AMP-forming, acetyl-CoA synthetase, to make ATP from acetyl-CoA. AMPforming, acetyl-CoA synthetases were previously thought to function only in the activation of acetate to acetyl-CoA.

Original languageEnglish (US)
Article numbere01208-16
JournalmBio
Volume7
Issue number4
DOIs
StatePublished - Jul 1 2016

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Acetyl Coenzyme A
Coenzymes
Acetate-CoA Ligase
Adenosine Triphosphate
Phosphate Acetyltransferase
Adenosine Monophosphate
Acetate Kinase
Acetates
Bacteria
Enzymes
Genome
diphosphoric acid
acetyladenosine triphosphate
Benzoates
Archaea
Proteome
Coculture Techniques
Cell Extracts
Transcriptome
Proteomics

ASJC Scopus subject areas

  • Microbiology
  • Virology

Cite this

James, K. L., Ríos-Hernández, L. A., Wofford, N. Q., Mouttaki, H., Sieber, J. R., Sheik, C. S., ... McInerney, M. J. (2016). Pyrophosphate-dependent ATP formation from acetyl coenzyme a in Syntrophus aciditrophicus, a new twist on ATP formation. mBio, 7(4), [e01208-16]. https://doi.org/10.1128/mBio.01208-16

Pyrophosphate-dependent ATP formation from acetyl coenzyme a in Syntrophus aciditrophicus, a new twist on ATP formation. / James, Kimberly L.; Ríos-Hernández, Luis A.; Wofford, Neil Q.; Mouttaki, Housna; Sieber, Jessica R.; Sheik, Cody S.; Nguyen, Hong H.; Yang, Yanan; Xie, Yongming; Erde, Jonathan; Rohlin, Lars; Karr, Elizabeth A.; Loo, Joseph A.; Loo, Rachel R.Ogorzalek; Hurst, Gregory B.; Gunsalus, Robert P.; Szweda, Luke I.; McInerney, Michael J.

In: mBio, Vol. 7, No. 4, e01208-16, 01.07.2016.

Research output: Contribution to journalArticle

James, KL, Ríos-Hernández, LA, Wofford, NQ, Mouttaki, H, Sieber, JR, Sheik, CS, Nguyen, HH, Yang, Y, Xie, Y, Erde, J, Rohlin, L, Karr, EA, Loo, JA, Loo, RRO, Hurst, GB, Gunsalus, RP, Szweda, LI & McInerney, MJ 2016, 'Pyrophosphate-dependent ATP formation from acetyl coenzyme a in Syntrophus aciditrophicus, a new twist on ATP formation', mBio, vol. 7, no. 4, e01208-16. https://doi.org/10.1128/mBio.01208-16
James KL, Ríos-Hernández LA, Wofford NQ, Mouttaki H, Sieber JR, Sheik CS et al. Pyrophosphate-dependent ATP formation from acetyl coenzyme a in Syntrophus aciditrophicus, a new twist on ATP formation. mBio. 2016 Jul 1;7(4). e01208-16. https://doi.org/10.1128/mBio.01208-16
James, Kimberly L. ; Ríos-Hernández, Luis A. ; Wofford, Neil Q. ; Mouttaki, Housna ; Sieber, Jessica R. ; Sheik, Cody S. ; Nguyen, Hong H. ; Yang, Yanan ; Xie, Yongming ; Erde, Jonathan ; Rohlin, Lars ; Karr, Elizabeth A. ; Loo, Joseph A. ; Loo, Rachel R.Ogorzalek ; Hurst, Gregory B. ; Gunsalus, Robert P. ; Szweda, Luke I. ; McInerney, Michael J. / Pyrophosphate-dependent ATP formation from acetyl coenzyme a in Syntrophus aciditrophicus, a new twist on ATP formation. In: mBio. 2016 ; Vol. 7, No. 4.
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abstract = "Syntrophus aciditrophicus is a model syntrophic bacterium that degrades key intermediates in anaerobic decomposition, such as benzoate, cyclohexane-1-carboxylate, and certain fatty acids, to acetate when grown with hydrogen-/formateconsuming microorganisms. ATP formation coupled to acetate production is the main source for energy conservation by S. aciditrophicus. However, the absence of homologs for phosphate acetyltransferase and acetate kinase in the genome of S. aciditrophicus leaves it unclear as to how ATP is formed, as most fermentative bacteria rely on these two enzymes to synthesize ATP from acetyl coenzyme A (CoA) and phosphate. Here, we combine transcriptomic, proteomic, metabolite, and enzymatic approaches to show that S. aciditrophicus uses AMP-forming, acetyl-CoA synthetase (Acs1) for ATP synthesis from acetyl- CoA. acs1 mRNA and Acs1 were abundant in transcriptomes and proteomes, respectively, of S. aciditrophicus grown in pure culture and coculture. Cell extracts of S. aciditrophicus had low or undetectable acetate kinase and phosphate acetyltransferase activities but had high acetyl-CoA synthetase activity under all growth conditions tested. Both Acs1 purified from S. aciditrophicus and recombinantly produced Acs1 catalyzed ATP and acetate formation from acetyl-CoA, AMP, and pyrophosphate. High pyrophosphate levels and a high AMP-to-ATP ratio (5.9 ± 1.4) in S. aciditrophicus cells support the operation of Acs1 in the acetate-forming direction. Thus, S. aciditrophicus has a unique approach to conserve energy involving pyrophosphate, AMP, acetyl-CoA, and an AMP-forming, acetyl-CoA synthetase. IMPORTANCE Bacteria use two enzymes, phosphate acetyltransferase and acetate kinase, to make ATP from acetyl-CoA, while acetate-forming archaea use a single enzyme, an ADP-forming, acetyl-CoA synthetase, to synthesize ATP and acetate from acetyl-CoA. Syntrophus aciditrophicus apparently relies on a different approach to conserve energy during acetyl-CoA metabolism, as its genome does not have homologs to the genes for phosphate acetyltransferase and acetate kinase. Here, we show that S. aciditrophicus uses an alternative approach, an AMP-forming, acetyl-CoA synthetase, to make ATP from acetyl-CoA. AMPforming, acetyl-CoA synthetases were previously thought to function only in the activation of acetate to acetyl-CoA.",
author = "James, {Kimberly L.} and R{\'i}os-Hern{\'a}ndez, {Luis A.} and Wofford, {Neil Q.} and Housna Mouttaki and Sieber, {Jessica R.} and Sheik, {Cody S.} and Nguyen, {Hong H.} and Yanan Yang and Yongming Xie and Jonathan Erde and Lars Rohlin and Karr, {Elizabeth A.} and Loo, {Joseph A.} and Loo, {Rachel R.Ogorzalek} and Hurst, {Gregory B.} and Gunsalus, {Robert P.} and Szweda, {Luke I.} and McInerney, {Michael J.}",
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AU - James, Kimberly L.

AU - Ríos-Hernández, Luis A.

AU - Wofford, Neil Q.

AU - Mouttaki, Housna

AU - Sieber, Jessica R.

AU - Sheik, Cody S.

AU - Nguyen, Hong H.

AU - Yang, Yanan

AU - Xie, Yongming

AU - Erde, Jonathan

AU - Rohlin, Lars

AU - Karr, Elizabeth A.

AU - Loo, Joseph A.

AU - Loo, Rachel R.Ogorzalek

AU - Hurst, Gregory B.

AU - Gunsalus, Robert P.

AU - Szweda, Luke I.

AU - McInerney, Michael J.

PY - 2016/7/1

Y1 - 2016/7/1

N2 - Syntrophus aciditrophicus is a model syntrophic bacterium that degrades key intermediates in anaerobic decomposition, such as benzoate, cyclohexane-1-carboxylate, and certain fatty acids, to acetate when grown with hydrogen-/formateconsuming microorganisms. ATP formation coupled to acetate production is the main source for energy conservation by S. aciditrophicus. However, the absence of homologs for phosphate acetyltransferase and acetate kinase in the genome of S. aciditrophicus leaves it unclear as to how ATP is formed, as most fermentative bacteria rely on these two enzymes to synthesize ATP from acetyl coenzyme A (CoA) and phosphate. Here, we combine transcriptomic, proteomic, metabolite, and enzymatic approaches to show that S. aciditrophicus uses AMP-forming, acetyl-CoA synthetase (Acs1) for ATP synthesis from acetyl- CoA. acs1 mRNA and Acs1 were abundant in transcriptomes and proteomes, respectively, of S. aciditrophicus grown in pure culture and coculture. Cell extracts of S. aciditrophicus had low or undetectable acetate kinase and phosphate acetyltransferase activities but had high acetyl-CoA synthetase activity under all growth conditions tested. Both Acs1 purified from S. aciditrophicus and recombinantly produced Acs1 catalyzed ATP and acetate formation from acetyl-CoA, AMP, and pyrophosphate. High pyrophosphate levels and a high AMP-to-ATP ratio (5.9 ± 1.4) in S. aciditrophicus cells support the operation of Acs1 in the acetate-forming direction. Thus, S. aciditrophicus has a unique approach to conserve energy involving pyrophosphate, AMP, acetyl-CoA, and an AMP-forming, acetyl-CoA synthetase. IMPORTANCE Bacteria use two enzymes, phosphate acetyltransferase and acetate kinase, to make ATP from acetyl-CoA, while acetate-forming archaea use a single enzyme, an ADP-forming, acetyl-CoA synthetase, to synthesize ATP and acetate from acetyl-CoA. Syntrophus aciditrophicus apparently relies on a different approach to conserve energy during acetyl-CoA metabolism, as its genome does not have homologs to the genes for phosphate acetyltransferase and acetate kinase. Here, we show that S. aciditrophicus uses an alternative approach, an AMP-forming, acetyl-CoA synthetase, to make ATP from acetyl-CoA. AMPforming, acetyl-CoA synthetases were previously thought to function only in the activation of acetate to acetyl-CoA.

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