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Title: Pyrophosphate-Dependent ATP Formation from Acetyl Coenzyme A in Syntrophus aciditrophicus, a New Twist on ATP Formation

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-/formate-consuming 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 Acs1more » 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.« less

Authors:
 [1];  [1];  [1];  [1];  [1];  [1];  [2];  [3];  [3];  [3];  [4];  [1];  [5];  [6];  [2];  [4];  [7]; ORCiD logo [1]
  1. Univ. of Oklahoma, Norman, OK (United States). Dept. of Microbiology and Plant Biology
  2. Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). Chemical Sciences Division
  3. Univ. of California, Los Angeles, CA (United States). Dept. of Chemistry and Biochemistry
  4. Univ. of California, Los Angeles, CA (United States). Dept. of Microbiology, Immunology and Molecular Genetics
  5. Univ. of California, Los Angeles, CA (United States). Dept. of Chemistry and Biochemistry; Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). Chemical Sciences Division
  6. Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). Chemical Sciences Division
  7. Oklahoma Medical Research Foundation, Oklahoma City, OK (United States). Aging and Metabolism Research Program
Publication Date:
Research Org.:
Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Univ. of Oklahoma, Norman, OK (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22). Chemical Sciences, Geosciences & Biosciences Division; USDOE Office of Science (SC), Biological and Environmental Research (BER); National Institutes of Health (NIH)
OSTI Identifier:
1324038
Alternate Identifier(s):
OSTI ID: 1604787
Grant/Contract Number:  
AC05-00OR22725; FG02-96ER20214; FC02-02ER63421; R01GM085402; FC02-02ER6342; FG02-08ER64689
Resource Type:
Journal Article: Accepted Manuscript
Journal Name:
mBio (Online)
Additional Journal Information:
Journal Volume: 7; Journal Issue: 4; Journal ID: ISSN 2150-7511
Publisher:
American Society for Microbiology
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; syntrophy; bioenergy; methanogenesis

Citation Formats

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., Ogorzalek Loo, Rachel R., Hurst, Gregory B., Gunsalus, Robert P., Szweda, Luke I., and McInerney, Michael J. Pyrophosphate-Dependent ATP Formation from Acetyl Coenzyme A in Syntrophus aciditrophicus, a New Twist on ATP Formation. United States: N. p., 2016. Web. doi: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., Ogorzalek Loo, Rachel R., 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. United States. 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., Ogorzalek Loo, Rachel R., Hurst, Gregory B., Gunsalus, Robert P., Szweda, Luke I., and McInerney, Michael J. Tue . "Pyrophosphate-Dependent ATP Formation from Acetyl Coenzyme A in Syntrophus aciditrophicus, a New Twist on ATP Formation". United States. https://doi.org/10.1128/mBio.01208-16. https://www.osti.gov/servlets/purl/1324038.
@article{osti_1324038,
title = {Pyrophosphate-Dependent ATP Formation from Acetyl Coenzyme A in Syntrophus aciditrophicus, a New Twist on ATP Formation},
author = {James, Kimberly L. and Ríos-Hernández, Luis A. and Wofford, Neil Q. and Mouttaki, Housna and Sieber, Jessica R. and Sheik, Cody S. and Nguyen, Hong H. and Yang, Yanan and Xie, Yongming and Erde, Jonathan and Rohlin, Lars and Karr, Elizabeth A. and Loo, Joseph A. and Ogorzalek Loo, Rachel R. and Hurst, Gregory B. and Gunsalus, Robert P. and Szweda, Luke I. and McInerney, Michael J.},
abstractNote = {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-/formate-consuming 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.},
doi = {10.1128/mBio.01208-16},
url = {https://www.osti.gov/biblio/1324038}, journal = {mBio (Online)},
issn = {2150-7511},
number = 4,
volume = 7,
place = {United States},
year = {2016},
month = {8}
}

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