Pyrophosphate-Dependent ATP Formation from Acetyl Coenzyme A in <named-content content-type="genus-species">Syntrophus aciditrophicus</named-content>, 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...
| Main Authors: | , , , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
American Society for Microbiology
2016-09-01
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| Series: | mBio |
| Online Access: | https://journals.asm.org/doi/10.1128/mBio.01208-16 |
| _version_ | 1818424638130094080 |
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| author | 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 |
| author_facet | 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 |
| author_sort | Kimberly L. James |
| collection | DOAJ |
| description | 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 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. AMP-forming, acetyl-CoA synthetases were previously thought to function only in the activation of acetate to acetyl-CoA. |
| first_indexed | 2024-12-14T14:01:13Z |
| format | Article |
| id | doaj.art-d1dab5dbda50462a851bf92528d0875e |
| institution | Directory Open Access Journal |
| issn | 2150-7511 |
| language | English |
| last_indexed | 2024-12-14T14:01:13Z |
| publishDate | 2016-09-01 |
| publisher | American Society for Microbiology |
| record_format | Article |
| series | mBio |
| spelling | doaj.art-d1dab5dbda50462a851bf92528d0875e2022-12-21T22:58:43ZengAmerican Society for MicrobiologymBio2150-75112016-09-017410.1128/mBio.01208-16Pyrophosphate-Dependent ATP Formation from Acetyl Coenzyme A in <named-content content-type="genus-species">Syntrophus aciditrophicus</named-content>, a New Twist on ATP FormationKimberly L. James0Luis A. Ríos-Hernández1Neil Q. Wofford2Housna Mouttaki3Jessica R. Sieber4Cody S. Sheik5Hong H. Nguyen6Yanan Yang7Yongming Xie8Jonathan Erde9Lars Rohlin10Elizabeth A. Karr11Joseph A. Loo12Rachel R. Ogorzalek Loo13Gregory B. Hurst14Robert P. Gunsalus15Luke I. Szweda16Michael J. McInerney17Department of Microbiology and Plant Biology, University of Oklahoma, Norman, Oklahoma, USADepartment of Microbiology and Plant Biology, University of Oklahoma, Norman, Oklahoma, USADepartment of Microbiology and Plant Biology, University of Oklahoma, Norman, Oklahoma, USADepartment of Microbiology and Plant Biology, University of Oklahoma, Norman, Oklahoma, USADepartment of Microbiology and Plant Biology, University of Oklahoma, Norman, Oklahoma, USADepartment of Microbiology and Plant Biology, University of Oklahoma, Norman, Oklahoma, USADepartment of Biological Chemistry, University of California Los Angeles, Los Angeles, California, USADepartment of Chemistry and Biochemistry, University of California Los Angeles, Los Angeles, California, USADepartment of Chemistry and Biochemistry, University of California Los Angeles, Los Angeles, California, USADepartment of Chemistry and Biochemistry, University of California Los Angeles, Los Angeles, California, USADepartment of Microbiology, Immunology and Molecular Genetics, University of California Los Angeles, Los Angeles, California, USADepartment of Microbiology and Plant Biology, University of Oklahoma, Norman, Oklahoma, USADepartment of Chemistry and Biochemistry, University of California Los Angeles, Los Angeles, California, USADepartment of Biological Chemistry, University of California Los Angeles, Los Angeles, California, USAChemical Sciences Division, Oak Ridge National Laboratory, Oak Ridge, Tennessee, USADepartment of Microbiology, Immunology and Molecular Genetics, University of California Los Angeles, Los Angeles, California, USAAging and Metabolism Research Program, Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma, USADepartment of Microbiology and Plant Biology, University of Oklahoma, Norman, Oklahoma, USAABSTRACT 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. 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. AMP-forming, acetyl-CoA synthetases were previously thought to function only in the activation of acetate to acetyl-CoA.https://journals.asm.org/doi/10.1128/mBio.01208-16 |
| spellingShingle | 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 Pyrophosphate-Dependent ATP Formation from Acetyl Coenzyme A in <named-content content-type="genus-species">Syntrophus aciditrophicus</named-content>, a New Twist on ATP Formation mBio |
| title | Pyrophosphate-Dependent ATP Formation from Acetyl Coenzyme A in <named-content content-type="genus-species">Syntrophus aciditrophicus</named-content>, a New Twist on ATP Formation |
| title_full | Pyrophosphate-Dependent ATP Formation from Acetyl Coenzyme A in <named-content content-type="genus-species">Syntrophus aciditrophicus</named-content>, a New Twist on ATP Formation |
| title_fullStr | Pyrophosphate-Dependent ATP Formation from Acetyl Coenzyme A in <named-content content-type="genus-species">Syntrophus aciditrophicus</named-content>, a New Twist on ATP Formation |
| title_full_unstemmed | Pyrophosphate-Dependent ATP Formation from Acetyl Coenzyme A in <named-content content-type="genus-species">Syntrophus aciditrophicus</named-content>, a New Twist on ATP Formation |
| title_short | Pyrophosphate-Dependent ATP Formation from Acetyl Coenzyme A in <named-content content-type="genus-species">Syntrophus aciditrophicus</named-content>, a New Twist on ATP Formation |
| title_sort | pyrophosphate dependent atp formation from acetyl coenzyme a in named content content type genus species syntrophus aciditrophicus named content a new twist on atp formation |
| url | https://journals.asm.org/doi/10.1128/mBio.01208-16 |
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