Identification and characterization of the tungsten-containing class of benzoyl-coenzyme A reductases

Kung, Johannes W.; Löffler, Claudia; Dörner, Katerina; Heintz, Dimitri; Gallien, Sébastien; Dorsselaer, Alain Van; Friedrich, Thorsten; Boll, Matthias

doi:10.1073/pnas.0905073106

Cited by 115 publications

(144 citation statements)

References 35 publications

(37 reference statements)

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“…It has been postulated that strict anaerobes must use ATP-independent class II BCRs because the consumption of two ATPs with class I BCRs would preclude a sufficient net gain of energy to support growth under strict anaerobic conditions (Kung et al, 2009Löffler et al, 2010). However, the fact that F. placidus is able to metabolize benzoate anoxically with an ATPconsuming class I BCR indicates that this rule does not apply to all anaerobic organisms and might be restricted to mesophilic bacteria.…”

Section: Discussionmentioning

confidence: 99%

“…The reaction catalyzed by this complex is reversible and ATP independent (Kung et al, 2009Lö ffler et al, 2010). It has been proposed that strict anaerobes must use class II BCRs because the amount of energy available from benzoate oxidation coupled to the reduction of Fe(III), sulfate or protons is not sufficient to support the substantial energetic requirement of the ATP-dependent class I BCR reaction (Schöcke and Schink, 1999;Kung et al, 2009Kung et al, , 2010Lö ffler et al, 2010).…”

Section: Introductionmentioning

confidence: 99%

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Genome-scale analysis of anaerobic benzoate and phenol metabolism in the hyperthermophilic archaeon Ferroglobus placidus

et al. 2011

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Insight into the mechanisms for the anaerobic metabolism of aromatic compounds by the hyperthermophilic archaeon Ferroglobus placidus is expected to improve understanding of the degradation of aromatics in hot (>80° C) environments and to identify enzymes that might have biotechnological applications. Analysis of the F. placidus genome revealed genes predicted to encode enzymes homologous to those previously identified as having a role in benzoate and phenol metabolism in mesophilic bacteria. Surprisingly, F. placidus lacks genes for an ATP-independent class II benzoyl-CoA (coenzyme A) reductase (BCR) found in all strictly anaerobic bacteria, but has instead genes coding for a bzd-type ATP-consuming class I BCR, similar to those found in facultative bacteria. The lower portion of the benzoate degradation pathway appears to be more similar to that found in the phototroph Rhodopseudomonas palustris, than the pathway reported for all heterotrophic anaerobic benzoate degraders. Many of the genes predicted to be involved in benzoate metabolism were found in one of two gene clusters. Genes for phenol carboxylation proceeding through a phenylphosphate intermediate were identified in a single gene cluster. Analysis of transcript abundance with a whole-genome microarray and quantitative reverse transcriptase polymerase chain reaction demonstrated that most of the genes predicted to be involved in benzoate or phenol metabolism had higher transcript abundance during growth on those substrates vs growth on acetate. These results suggest that the general strategies for benzoate and phenol metabolism are highly conserved between microorganisms living in moderate and hot environments, and that anaerobic metabolism of aromatic compounds might be analyzed in a wide range of environments with similar molecular targets.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Genome-scale analysis of anaerobic benzoate and phenol metabolism in the hyperthermophilic archaeon Ferroglobus placidus

et al. 2011

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“…Instead, homologs are found in the alternative BCRindependent mechanism within a set of 44 genes that have been postulated to operate in Geobacter metallireducens and 'Syntrophus aciditrophicus' (Butler et al, 2007;McInerney et al, 2007;Peters et al, 2007). The key BCR enzyme in G. metallireducens was successfully characterized in vitro (Kung et al, 2009). The metagenome analysis further predicted the pathways that are used for conversion of benzoate to hydroxypimelyl-CoA and subsequent conversion of 3-hydroxypimelyl-CoA via b-oxidation to acetyl-CoA, which in turn gives rise to acetate through substrate-level phosphorylation (Supplementary Figure 7; Figure 3).…”

Section: Pelotomaculummentioning

confidence: 99%

Multiple syntrophic interactions in a terephthalate-degrading methanogenic consortium

et al. 2010

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Terephthalate (TA) is one of the top 50 chemicals produced worldwide. Its production results in a TA-containing wastewater that is treated by anaerobic processes through a poorly understood methanogenic syntrophy. Using metagenomics, we characterized the methanogenic consortium inside a hyper-mesophilic (that is, between mesophilic and thermophilic), TA-degrading bioreactor. We identified genes belonging to dominant Pelotomaculum species presumably involved in TA degradation through decarboxylation, dearomatization, and modified b-oxidation to H 2 /CO 2 and acetate. These intermediates are converted to CH 4 /CO 2 by three novel hyper-mesophilic methanogens. Additional secondary syntrophic interactions were predicted in Thermotogae, Syntrophus and candidate phyla OP5 and WWE1 populations. The OP5 encodes genes capable of anaerobic autotrophic butyrate production and Thermotogae, Syntrophus and WWE1 have the genetic potential to oxidize butyrate to CO 2 /H 2 and acetate. These observations suggest that the TA-degrading consortium consists of additional syntrophic interactions beyond the standard H 2 -producing syntroph-methanogen partnership that may serve to improve community stability. The ISME Journal (2011) 5, 122-130; doi:10.1038/ismej.2010; published online 5 August 2010Subject Category: integrated genomics and post-genomics approaches in microbial ecology Keywords: metagenomics; methanogenesis; syntroph; microbial diversity; carbon cycling Introduction Terephthalate (TA) is used as the raw material for the manufacture of numerous plastic products (for example, polyethylene TA bottles and textile fibers). During its production, TA-containing wastewater is discharged in large volumes (as high as 300 million m 3 per year) and high concentration (up to 20 kg COD (chemical oxygen demand) m À3 ) (Razo-Flores et al., 2006). This wastewater is generally treated by anaerobic biological processes under mesophilic conditions (B35 1C). However, anaerobic processes operated at hyper-mesophilic (46-50 1C) and thermophilic (B55 1C) temperatures may be preferable because of the ability to achieve higher loading rate (van Lier et al., 1997;Chen et al., 2004), which reduces the reactor volume. Moreover, TA wastewater is usually generated at 54-60 1C, and does not require additional energy input for maintaining reactor temperature (Chen et al., 2004). The microbial biomass usually occurs in the form of granules or biofilms attaching on the surface of porous media. Under such environments, TA degradation has been hypothesized (Kleerebezem et al., 1999) to be based on a syntrophic microbial relationship whereby fermentative H 2 -producing bacteria (syntrophs) convert TA through benzoate to acetate and H 2 /CO 2 , and acetoclastic and hydrogenotrophic methanogens further convert the intermediates to methane by physically positioning themselves close to the syntrophs to overcome the www.nature.com/ismej thermodynamic barrier (Stams, 1994;Conrad, 1999;Dolfing, 2001).In practice, the complexities of TA-degrading communities are not...

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“…Cell extracts of S. aciditrophicus converted Ch1CoA to Ch1,5CoA (0.6 mol min Ϫ1 mg Ϫ1 ), which was subsequently converted to benzoyl-CoA (0.2 mol min Ϫ1 mg Ϫ1 ) in a time-, enzyme-, and ferricenium hexafluorophosphate-dependent reaction. While the formation of Ch1,5CoA was expected to be catalyzed by a previously unknown Ch1CoA dehydrogenase, the further conversion of Ch1,5CoA to benzoyl-CoA could also be assigned to the reverse activity of the class II benzoyl-CoA reductases in the cell extract, as described previously for the Geobacter metallireducens enzyme (15). Attempts to purify the Ch1CoA-oxidizing enzyme from cell extracts of S. aciditrophicus failed due to highly similar chromatographic binding properties for other proteins (e.g., ChCoA dehydrogenase and subunits of benzoyl-CoA reductase).…”

Section: General Remarksmentioning

confidence: 99%

Cyclohexanecarboxyl-Coenzyme A (CoA) and Cyclohex-1-ene-1-Carboxyl-CoA Dehydrogenases, Two Enzymes Involved in the Fermentation of Benzoate and Crotonate in Syntrophus aciditrophicus

Kung¹,

Seifert²,

Bergen³

et al. 2013

Journal of Bacteriology

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The strictly anaerobic Syntrophus aciditrophicus is a fermenting deltaproteobacterium that is able to degrade benzoate or crotonate in the presence and in the absence of a hydrogen-consuming partner. During growth in pure culture, both substrates are dismutated to acetate and cyclohexane carboxylate. In this work, the unknown enzymes involved in the late steps of cyclohexane carboxylate formation were studied. Using enzyme assays monitoring the oxidative direction, a cyclohex-1-ene-1-carboxyl-CoA (Ch1CoA)-forming cyclohexanecarboxyl-CoA (ChCoA) dehydrogenase was purified and characterized from S. aciditrophicus and after heterologous expression of its gene in Escherichia coli. In addition, a cyclohexa-1,5-diene-1-carboxyl-CoA (Ch1,5CoA)-forming Ch1CoA dehydrogenase was characterized after purification of the heterologously expressed gene. Both enzymes had a native molecular mass of 150 kDa and were composed of a single, 40-to 45-kDa subunit; both contained flavin adenine dinucleotide (FAD) as a cofactor. While the ChCoA dehydrogenase was competitively inhibited by Ch1CoA in the oxidative direction, Ch1CoA dehydrogenase further converted the product Ch1,5CoA to benzoyl-CoA. The results obtained suggest that Ch1,5CoA is a common intermediate in benzoate and crotonate fermentation that serves as an electron-accepting substrate for the two consecutively operating acyl-CoA dehydrogenases characterized in this work. In the case of benzoate fermentation, Ch1,5CoA is formed by a class II benzoyl-CoA reductase; in the case of crotonate fermentation, Ch1,5CoA is formed by reversing the reactions of the benzoyl-CoA degradation pathway that are also employed during the oxidative (degradative) branch of benzoate fermentation.

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Identification and characterization of the tungsten-containing class of benzoyl-coenzyme A reductases

Abstract: anaerobic aromatic degradation ͉ AOR ͉ geobacter

Cited by 115 publications

References 35 publications

Genome-scale analysis of anaerobic benzoate and phenol metabolism in the hyperthermophilic archaeon Ferroglobus placidus

Genome-scale analysis of anaerobic benzoate and phenol metabolism in the hyperthermophilic archaeon Ferroglobus placidus

Multiple syntrophic interactions in a terephthalate-degrading methanogenic consortium

Cyclohexanecarboxyl-Coenzyme A (CoA) and Cyclohex-1-ene-1-Carboxyl-CoA Dehydrogenases, Two Enzymes Involved in the Fermentation of Benzoate and Crotonate in Syntrophus aciditrophicus

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