Benzylsuccinate Synthase of
            <i>Azoarcus</i>
            sp. Strain T: Cloning, Sequencing, Transcriptional Organization, and Its Role in Anaerobic Toluene and
            <i>m</i>
            -Xylene Mineralization

Achong, Gypsy R.; Rodrı́guez, Ana M.; Spormann, Alfred M.

doi:10.1128/jb.183.23.6763-6770.2001

Cited by 94 publications

(92 citation statements)

References 35 publications

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“…The bacterium used the same enzyme for addition of fumarate to m-xylene and to toluene. A mutant deleted in bss, which codes for benzylsuccinate synthase, was unable to grow on either substrate, but grew on benzoate (Achong et al 2001). After the gene was restored, the capability of the denitrifying bacterium to initiate m-xylene and toluene degradation was recuperated.…”

Section: Discussionmentioning

confidence: 90%

Degradation of o -xylene and m -xylene by a novel sulfate-reducer belonging to the genus Desulfotomaculum

Morasch

Schink

Tebbe

et al. 2004

Archives of Microbiology

118

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A strictly anaerobic bacterium, strain OX39, was isolated with o-xylene as organic substrate and sulfate as electron acceptor from an aquifer at a former gasworks plant contaminated with aromatic hydrocarbons. Apart from o-xylene, strain OX39 grew on m-xylene and toluene and all three substrates were oxidized completely to CO 2 . Induction experiments indicated that o-xylene, m-xylene, and toluene degradation were initiated by different specific enzymes. Methylbenzylsuccinate was identified in supernatants of cultures grown on o-xylene and m-xylene, and benzylsuccinate was detected in supernatants of toluene-grown cells, thus indicating that degradation was initiated in all three cases by fumarate addition to the methyl group. Strain OX39 was sensitive towards sulfide and depended on Fe(II) in the medium as a scavenger of the produced sulfide. Analysis of the PCR-amplified 16S rRNA gene revealed that strain OX39 affiliates with the gram-positive endospore-forming sulfate reducers of the genus Desulfotomaculum and is the first hydrocarbonoxidizing bacterium in this genus. Keywords Sulfate reduction · BTEX · Benzylsuccinate synthase · Desulfotomaculum IntroductionPetroleum hydrocarbons are among the most abundant groundwater contaminants and can be found in gasworks plants, landfill leachates, and accidental fuel spills (US-EPA 1999). The mono-aromatic hydrocarbons benzene, toluene, ethylbenzene, and xylene isomers (BTEX) are putatively mutagenic or carcinogenic substances and make-up more than 50% by weight of the water-soluble gasoline fraction (Coleman et al. 1984). Due to their relatively high solubility, they are mobile with the groundwater flow and form contaminant plumes in aquifers. The degradative potential of anaerobic bacteria towards aromatic hydrocarbons in situ was investigated in several studies and a decrease of BTEX compounds could be demonstrated under denitrifying, Fe(III)-reducing, sulfate-reducing, and methanogenic conditions (Dolfing et al. 1990;Kazumi et al. 1997;Reinhard et al. 1997;Gieg et al. 1999;Phelps and Young 2001). Numerous anaerobic bacterial cultures have been enriched in the past, and pure strains have been isolated. Of all BTEX compounds, toluene has been studied most extensively with respect to its anaerobic degradation. Pure cultures of toluene-degrading bacteria that use NO 3 -, Fe(III), or SO 4 2-as electron acceptors have been isolated (Lovley and Lonergan 1990;Rabus et al. 1993;Seyfried et al. 1994;Zhou et al. 1995;Beller et al. 1996). Under anoxic conditions, degradation of toluene proceeds via addition of fumarate to the methyl group (Biegert et al. 1996;Beller and Spormann 1997b;Leuthner et al. 1998;Rabus and Heider 1998;Kane et al. 2002).Consumption of o-xylene was discovered simultaneously with toluene degradation in methanogenic consortia (Edwards and Grbic-Galic 1994). Later, transformation of o-xylene to o-methyl homologs of benzylsuccinate by toluene-degrading strains was reported; however, no further oxidation steps could be observed (Beller and Spormann...

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Section: Discussionmentioning

confidence: 90%

Degradation of o -xylene and m -xylene by a novel sulfate-reducer belonging to the genus Desulfotomaculum

Morasch

Schink

Tebbe

et al. 2004

Archives of Microbiology

118

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“…However, in Azoarcus sp. strain T, the bssF gene appears to have its own transcriptional start site (1). Comparative analysis reveals that the order of the genes in the bssDCABEF cluster is conserved in all toluenedegrading bacteria analyzed so far (Fig.…”

Section: Gene Clusters For Degradation Of Aromatic Hydrocarbonsmentioning

confidence: 99%

“…Gene inactivation studies with Azoarcus sp. strain T and T. aromatica strain T1 revealed that each of the subunits is essential for the function of the benzylsuccinate synthase (1,69). Site-directed mutagenesis of the tutF, tutD, and tutG genes from T. aromatica strain T1 determined residues that are critical for the activity of benzylsuccinate synthase (23,72).…”

Section: Gene Clusters For Degradation Of Aromatic Hydrocarbonsmentioning

confidence: 99%

“…On the other hand, upstream of the bss clusters in Azoarcus and Thauera strains, there are two adjacent genes, tdiR (tutB1) and tdiS (tutC1) (Fig. 16), that code for a putative two-component regulatory system that was suggested to control the expression of the bss and bbs genes (see below) (1,202). Orthologs of tdiSR are lacking in G. metallireducens, and putative xylR-like and tetR-like regulatory genes are flanking the bss and bbs clusters, respectively (Fig.…”

Section: Vol 73 2009 Anaerobic Biodegradation Of Aromatics 101mentioning

confidence: 99%

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Anaerobic Catabolism of Aromatic Compounds: a Genetic and Genomic View

Carmona

Zamarro

Blázquez

et al. 2009

Microbiol Mol Biol Rev

392

381

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SUMMARY Aromatic compounds belong to one of the most widely distributed classes of organic compounds in nature, and a significant number of xenobiotics belong to this family of compounds. Since many habitats containing large amounts of aromatic compounds are often anoxic, the anaerobic catabolism of aromatic compounds by microorganisms becomes crucial in biogeochemical cycles and in the sustainable development of the biosphere. The mineralization of aromatic compounds by facultative or obligate anaerobic bacteria can be coupled to anaerobic respiration with a variety of electron acceptors as well as to fermentation and anoxygenic photosynthesis. Since the redox potential of the electron-accepting system dictates the degradative strategy, there is wide biochemical diversity among anaerobic aromatic degraders. However, the genetic determinants of all these processes and the mechanisms involved in their regulation are much less studied. This review focuses on the recent findings that standard molecular biology approaches together with new high-throughput technologies (e.g., genome sequencing, transcriptomics, proteomics, and metagenomics) have provided regarding the genetics, regulation, ecophysiology, and evolution of anaerobic aromatic degradation pathways. These studies revealed that the anaerobic catabolism of aromatic compounds is more diverse and widespread than previously thought, and the complex metabolic and stress programs associated with the use of aromatic compounds under anaerobic conditions are starting to be unraveled. Anaerobic biotransformation processes based on unprecedented enzymes and pathways with novel metabolic capabilities, as well as the design of novel regulatory circuits and catabolic networks of great biotechnological potential in synthetic biology, are now feasible to approach.

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“…strain M5; 19 and the TdiSR system regulating benzylsuccinate synthase which is involved in the anaerobic catabolism of toluene in Azoarcus sp. strain T. 1 Members of these TCSs, and others, have been incorporated into the rational design of biosensors by integrating the sensing apparatus with appropriate reporter genes (e.g., lux, gfp) for in situ monitoring of aromatic pollutant presence in both aquatic and soil environments. 33 The application of such biosensors is not without challenges however, as they are typically characterized under laboratory conditions that are often not reflective of realtime settings.…”

Section: Introductionmentioning

confidence: 99%

Functional characterization of a StyS sensor kinase reveals distinct domains associated with intracellular and extracellular sensing of styrene inP. putidaCA-3

O’Leary¹,

Mooney

O’Mahony³

et al. 2014

Bioengineered

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Benzylsuccinate Synthase of Azoarcus sp. Strain T: Cloning, Sequencing, Transcriptional Organization, and Its Role in Anaerobic Toluene and m -Xylene Mineralization

Cited by 94 publications

References 35 publications

Degradation of o -xylene and m -xylene by a novel sulfate-reducer belonging to the genus Desulfotomaculum

Degradation of o -xylene and m -xylene by a novel sulfate-reducer belonging to the genus Desulfotomaculum

Anaerobic Catabolism of Aromatic Compounds: a Genetic and Genomic View

Functional characterization of a StyS sensor kinase reveals distinct domains associated with intracellular and extracellular sensing of styrene inP. putidaCA-3

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