Activation of short-chain ketones and isopropanol in sulfate-reducing bacteria

Abstract Background Degradation of acetone by aerobic and nitrate-reducing bacteria can proceed via carboxylation to acetoacetate and subsequent thiolytic cleavage to two acetyl residues. A different strategy was identified in the sulfate-reducing bacterium Desulfococcus biacutus that involves formy...

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Main Authors: Jasmin Frey, Sophie Kaßner, Dieter Spiteller, Mario Mergelsberg, Matthias Boll, David Schleheck, Bernhard Schink
Format: Article
Language:English
Published: BMC 2021-02-01
Series:BMC Microbiology
Subjects:
Online Access:https://doi.org/10.1186/s12866-021-02112-6
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spelling doaj-6a2fba4a236442c3b0936f112391fb5d2021-02-21T12:20:56ZengBMCBMC Microbiology1471-21802021-02-0121111110.1186/s12866-021-02112-6Activation of short-chain ketones and isopropanol in sulfate-reducing bacteriaJasmin Frey0Sophie Kaßner1Dieter Spiteller2Mario Mergelsberg3Matthias Boll4David Schleheck5Bernhard Schink6Department of Biology, University of KonstanzDepartment of Biology, University of KonstanzDepartment of Biology, University of KonstanzInstitute of Biology, Albert-Ludwigs-UniversitätInstitute of Biology, Albert-Ludwigs-UniversitätDepartment of Biology, University of KonstanzDepartment of Biology, University of KonstanzAbstract Background Degradation of acetone by aerobic and nitrate-reducing bacteria can proceed via carboxylation to acetoacetate and subsequent thiolytic cleavage to two acetyl residues. A different strategy was identified in the sulfate-reducing bacterium Desulfococcus biacutus that involves formylation of acetone to 2-hydroxyisobutyryl-CoA. Results Utilization of short-chain ketones (acetone, butanone, 2-pentanone and 3-pentanone) and isopropanol by the sulfate reducer Desulfosarcina cetonica was investigated by differential proteome analyses and enzyme assays. Two-dimensional protein gel electrophoresis indicated that D. cetonica during growth with acetone expresses enzymes homologous to those described for Desulfococcus biacutus: a thiamine diphosphate (TDP)-requiring enzyme, two subunits of a B12-dependent mutase, and a NAD+-dependent dehydrogenase. Total proteomics of cell-free extracts confirmed these results and identified several additional ketone-inducible proteins. Acetone is activated, most likely mediated by the TDP-dependent enzyme, to a branched-chain CoA-ester, 2-hydroxyisobutyryl-CoA. This compound is linearized to 3-hydroxybutyryl-CoA by a coenzyme B12-dependent mutase followed by oxidation to acetoacetyl-CoA by a dehydrogenase. Proteomic analysis of isopropanol- and butanone-grown cells revealed the expression of a set of enzymes identical to that expressed during growth with acetone. Enzyme assays with cell-free extract of isopropanol- and butanone-grown cells support a B12-dependent isomerization. After growth with 2-pentanone or 3-pentanone, similar protein patterns were observed in cell-free extracts as those found after growth with acetone. Conclusions According to these results, butanone and isopropanol, as well as the two pentanone isomers, are degraded by the same enzymes that are used also in acetone degradation. Our results indicate that the degradation of several short-chain ketones appears to be initiated by TDP-dependent formylation in sulfate-reducing bacteria.https://doi.org/10.1186/s12866-021-02112-6Anaerobic acetone degradationKetone degradationPentanoneSulfate reduction2-hydroxyisobutyryl-CoAThiamine diphosphate
collection DOAJ
language English
format Article
sources DOAJ
author Jasmin Frey
Sophie Kaßner
Dieter Spiteller
Mario Mergelsberg
Matthias Boll
David Schleheck
Bernhard Schink
spellingShingle Jasmin Frey
Sophie Kaßner
Dieter Spiteller
Mario Mergelsberg
Matthias Boll
David Schleheck
Bernhard Schink
Activation of short-chain ketones and isopropanol in sulfate-reducing bacteria
BMC Microbiology
Anaerobic acetone degradation
Ketone degradation
Pentanone
Sulfate reduction
2-hydroxyisobutyryl-CoA
Thiamine diphosphate
author_facet Jasmin Frey
Sophie Kaßner
Dieter Spiteller
Mario Mergelsberg
Matthias Boll
David Schleheck
Bernhard Schink
author_sort Jasmin Frey
title Activation of short-chain ketones and isopropanol in sulfate-reducing bacteria
title_short Activation of short-chain ketones and isopropanol in sulfate-reducing bacteria
title_full Activation of short-chain ketones and isopropanol in sulfate-reducing bacteria
title_fullStr Activation of short-chain ketones and isopropanol in sulfate-reducing bacteria
title_full_unstemmed Activation of short-chain ketones and isopropanol in sulfate-reducing bacteria
title_sort activation of short-chain ketones and isopropanol in sulfate-reducing bacteria
publisher BMC
series BMC Microbiology
issn 1471-2180
publishDate 2021-02-01
description Abstract Background Degradation of acetone by aerobic and nitrate-reducing bacteria can proceed via carboxylation to acetoacetate and subsequent thiolytic cleavage to two acetyl residues. A different strategy was identified in the sulfate-reducing bacterium Desulfococcus biacutus that involves formylation of acetone to 2-hydroxyisobutyryl-CoA. Results Utilization of short-chain ketones (acetone, butanone, 2-pentanone and 3-pentanone) and isopropanol by the sulfate reducer Desulfosarcina cetonica was investigated by differential proteome analyses and enzyme assays. Two-dimensional protein gel electrophoresis indicated that D. cetonica during growth with acetone expresses enzymes homologous to those described for Desulfococcus biacutus: a thiamine diphosphate (TDP)-requiring enzyme, two subunits of a B12-dependent mutase, and a NAD+-dependent dehydrogenase. Total proteomics of cell-free extracts confirmed these results and identified several additional ketone-inducible proteins. Acetone is activated, most likely mediated by the TDP-dependent enzyme, to a branched-chain CoA-ester, 2-hydroxyisobutyryl-CoA. This compound is linearized to 3-hydroxybutyryl-CoA by a coenzyme B12-dependent mutase followed by oxidation to acetoacetyl-CoA by a dehydrogenase. Proteomic analysis of isopropanol- and butanone-grown cells revealed the expression of a set of enzymes identical to that expressed during growth with acetone. Enzyme assays with cell-free extract of isopropanol- and butanone-grown cells support a B12-dependent isomerization. After growth with 2-pentanone or 3-pentanone, similar protein patterns were observed in cell-free extracts as those found after growth with acetone. Conclusions According to these results, butanone and isopropanol, as well as the two pentanone isomers, are degraded by the same enzymes that are used also in acetone degradation. Our results indicate that the degradation of several short-chain ketones appears to be initiated by TDP-dependent formylation in sulfate-reducing bacteria.
topic Anaerobic acetone degradation
Ketone degradation
Pentanone
Sulfate reduction
2-hydroxyisobutyryl-CoA
Thiamine diphosphate
url https://doi.org/10.1186/s12866-021-02112-6
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