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Membrane Complexes of Syntrophomonas wolfei Involved in Syntrophic Butyrate Degradation and Hydrogen Formation

Syntrophic butyrate metabolism involves the thermodynamically unfavorable production of hydrogen and/or formate from the high potential electron donor, butyryl-CoA. Such redox reactions can occur only with energy input by a process called reverse electron transfer. Previous studies have demonstrated...

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Autores principales: Crable, Bryan R., Sieber, Jessica R., Mao, Xinwei, Alvarez-Cohen, Lisa, Gunsalus, Robert, Ogorzalek Loo, Rachel R., Nguyen, Hong, McInerney, Michael J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5101538/
https://www.ncbi.nlm.nih.gov/pubmed/27881975
http://dx.doi.org/10.3389/fmicb.2016.01795
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author Crable, Bryan R.
Sieber, Jessica R.
Mao, Xinwei
Alvarez-Cohen, Lisa
Gunsalus, Robert
Ogorzalek Loo, Rachel R.
Nguyen, Hong
McInerney, Michael J.
author_facet Crable, Bryan R.
Sieber, Jessica R.
Mao, Xinwei
Alvarez-Cohen, Lisa
Gunsalus, Robert
Ogorzalek Loo, Rachel R.
Nguyen, Hong
McInerney, Michael J.
author_sort Crable, Bryan R.
collection PubMed
description Syntrophic butyrate metabolism involves the thermodynamically unfavorable production of hydrogen and/or formate from the high potential electron donor, butyryl-CoA. Such redox reactions can occur only with energy input by a process called reverse electron transfer. Previous studies have demonstrated that hydrogen production from butyrate requires the presence of a proton gradient, but the biochemical machinery involved has not been clearly elucidated. In this study, the gene and enzyme systems involved in reverse electron transfer by Syntrophomonas wolfei were investigated using proteomic and gene expression approaches. S. wolfei was grown in co-culture with Methanospirillum hungatei or Dehalococcoides mccartyi under conditions requiring reverse electron transfer and compared to both axenic S. wolfei cultures and co-cultures grown in conditions that do not require reverse electron transfer. Blue native gel analysis of membranes solubilized from syntrophically grown cells revealed the presence of a membrane-bound hydrogenase, Hyd2, which exhibited hydrogenase activity during in gel assays. Bands containing a putative iron-sulfur (FeS) oxidoreductase were detected in membranes of crotonate-grown and butyrate grown S. wolfei cells. The genes for the corresponding hydrogenase subunits, hyd2ABC, were differentially expressed at higher levels during syntrophic butyrate growth when compared to growth on crotonate. The expression of the FeS oxidoreductase gene increased when S. wolfei was grown with M. hungatei. Additional membrane-associated proteins detected included F(o)F(1) ATP synthase subunits and several membrane transporters that may aid syntrophic growth. Furthermore, syntrophic butyrate metabolism can proceed exclusively by interspecies hydrogen transfer, as demonstrated by growth with D. mccartyi, which is unable to use formate. These results argue for the importance of Hyd2 and FeS oxidoreductase in reverse electron transfer during syntrophic butyrate degradation.
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spelling pubmed-51015382016-11-23 Membrane Complexes of Syntrophomonas wolfei Involved in Syntrophic Butyrate Degradation and Hydrogen Formation Crable, Bryan R. Sieber, Jessica R. Mao, Xinwei Alvarez-Cohen, Lisa Gunsalus, Robert Ogorzalek Loo, Rachel R. Nguyen, Hong McInerney, Michael J. Front Microbiol Microbiology Syntrophic butyrate metabolism involves the thermodynamically unfavorable production of hydrogen and/or formate from the high potential electron donor, butyryl-CoA. Such redox reactions can occur only with energy input by a process called reverse electron transfer. Previous studies have demonstrated that hydrogen production from butyrate requires the presence of a proton gradient, but the biochemical machinery involved has not been clearly elucidated. In this study, the gene and enzyme systems involved in reverse electron transfer by Syntrophomonas wolfei were investigated using proteomic and gene expression approaches. S. wolfei was grown in co-culture with Methanospirillum hungatei or Dehalococcoides mccartyi under conditions requiring reverse electron transfer and compared to both axenic S. wolfei cultures and co-cultures grown in conditions that do not require reverse electron transfer. Blue native gel analysis of membranes solubilized from syntrophically grown cells revealed the presence of a membrane-bound hydrogenase, Hyd2, which exhibited hydrogenase activity during in gel assays. Bands containing a putative iron-sulfur (FeS) oxidoreductase were detected in membranes of crotonate-grown and butyrate grown S. wolfei cells. The genes for the corresponding hydrogenase subunits, hyd2ABC, were differentially expressed at higher levels during syntrophic butyrate growth when compared to growth on crotonate. The expression of the FeS oxidoreductase gene increased when S. wolfei was grown with M. hungatei. Additional membrane-associated proteins detected included F(o)F(1) ATP synthase subunits and several membrane transporters that may aid syntrophic growth. Furthermore, syntrophic butyrate metabolism can proceed exclusively by interspecies hydrogen transfer, as demonstrated by growth with D. mccartyi, which is unable to use formate. These results argue for the importance of Hyd2 and FeS oxidoreductase in reverse electron transfer during syntrophic butyrate degradation. Frontiers Media S.A. 2016-11-09 /pmc/articles/PMC5101538/ /pubmed/27881975 http://dx.doi.org/10.3389/fmicb.2016.01795 Text en Copyright © 2016 Crable, Sieber, Mao, Alvarez-Cohen, Gunsalus, Ogorzalek Loo, Nguyen and McInerney. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Microbiology
Crable, Bryan R.
Sieber, Jessica R.
Mao, Xinwei
Alvarez-Cohen, Lisa
Gunsalus, Robert
Ogorzalek Loo, Rachel R.
Nguyen, Hong
McInerney, Michael J.
Membrane Complexes of Syntrophomonas wolfei Involved in Syntrophic Butyrate Degradation and Hydrogen Formation
title Membrane Complexes of Syntrophomonas wolfei Involved in Syntrophic Butyrate Degradation and Hydrogen Formation
title_full Membrane Complexes of Syntrophomonas wolfei Involved in Syntrophic Butyrate Degradation and Hydrogen Formation
title_fullStr Membrane Complexes of Syntrophomonas wolfei Involved in Syntrophic Butyrate Degradation and Hydrogen Formation
title_full_unstemmed Membrane Complexes of Syntrophomonas wolfei Involved in Syntrophic Butyrate Degradation and Hydrogen Formation
title_short Membrane Complexes of Syntrophomonas wolfei Involved in Syntrophic Butyrate Degradation and Hydrogen Formation
title_sort membrane complexes of syntrophomonas wolfei involved in syntrophic butyrate degradation and hydrogen formation
topic Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5101538/
https://www.ncbi.nlm.nih.gov/pubmed/27881975
http://dx.doi.org/10.3389/fmicb.2016.01795
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