Site-directed mutagenesis of the Thauera aromatica strain T1 tutE tutFDGH gene cluster

Bhandare, Reena; Calabro, Mark; Coschigano, Peter W.

doi:10.1016/j.bbrc.2006.05.199

Cited by 8 publications

(13 citation statements)

References 38 publications

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“…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). The bssD (tutE) gene, which is always located upstream of the bssCAB (tutFDG) genes, codes for BssD (TutE) activase, which introduces the glycyl radical at the large subunit (BssA/ TutD) of benzylsuccinate synthase in an S-adenosylmethionine-dependent reaction (332).…”

Section: Gene Clusters For Degradation Of Aromatic Hydrocarbonsmentioning

confidence: 99%

“…The bssD (tutE) gene, which is always located upstream of the bssCAB (tutFDG) genes, codes for BssD (TutE) activase, which introduces the glycyl radical at the large subunit (BssA/ TutD) of benzylsuccinate synthase in an S-adenosylmethionine-dependent reaction (332). Site-directed mutagenesis of certain cysteine residues of TutE in T. aromatica strain T1 revealed that they are likely to be critical for the radical activation of benzylsuccinate synthase (23). The benzylsuccinate synthase is irreversibly inactivated in the presence of molecular oxygen by the oxygenolytic cleavage of the ␣-subunit (200,217).…”

Section: Gene Clusters For Degradation Of Aromatic Hydrocarbonsmentioning

confidence: 99%

“…The bssE gene product contains a Walker-type ATP/GTP binding-site motif and is similar to an emerging class of chaperone-like ATPases required for the assembly, operation, and disassembly of protein complexes, which suggests that it may be required for either the assembly or activation of benzylsuccinate synthase (33,155). The tutH gene product in T. aromatica strain T1 has been shown to be necessary for toluene metabolism, and the 52-Gly Lys Ser-54 residues, which are part of the putative ATP/GTP binding domain, are essential for its activity (23). The bssF gene was predicted to be part of the bss operon because genes coding for highly similar proteins are found directly downstream of bssE in all completely sequenced bss clusters (Fig.…”

Section: Gene Clusters For Degradation Of Aromatic Hydrocarbonsmentioning

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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Section: Gene Clusters For Degradation Of Aromatic Hydrocarbonsmentioning

confidence: 99%

Section: Gene Clusters For Degradation Of Aromatic Hydrocarbonsmentioning

confidence: 99%

Section: Gene Clusters For Degradation Of Aromatic Hydrocarbonsmentioning

confidence: 99%

See 1 more Smart Citation

Anaerobic Catabolism of Aromatic Compounds: a Genetic and Genomic View

Carmona

Zamarro

Blázquez

et al. 2009

Microbiol Mol Biol Rev

392

381

View full text Add to dashboard Cite

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“…BSS␣ contains the 10-stranded barrel and is the catalytic subunit, whereas BSS␤ and BSS␥ are both small, [4Fe-4S]-cluster-containing proteins of unknown function (19,20). Despite the fact that the roles of BSS␤ and BSS␥ are unclear, both proteins are required for viable growth of T. aromatica strain T1 on toluene (21,22). We previously solved structures of the BSS␣␤␥ and BSS␣␥ complexes, revealing that removal of BSS␤ from the complex with BSS␣␥ permits a large conformational change within BSS␣ (23).…”

mentioning

confidence: 99%

Substrate-bound Structures of Benzylsuccinate Synthase Reveal How Toluene Is Activated in Anaerobic Hydrocarbon Degradation

Funk

Marsh

Drennan

2015

Journal of Biological Chemistry

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Background: Benzylsuccinate synthase (BSS) catalyzes the formation of a COC bond between toluene and fumarate by a radical mechanism. Results: BSS binds substrates in a buried active site and uses conformational changes to gate access. Conclusion: BSS and related enzymes precisely position substrates for chemistry. Significance: These structures reveal the molecular basis for BSS radical chemistry and substrate specificity.

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“…denitrificans cannot not grow on the media solidified with agar or gelrite, therefore preventing the use of conventional molecular biological techniques. Molecular biology experiments have been performed using various Thauera strains (Bhandare et al, 2006). Thus, T. terpenica 58Eu can be used as another model organism for anaerobic steroid degradation.…”

Section: Discussionmentioning

confidence: 99%

Integrated multi-omics analyses reveal the biochemical mechanisms and phylogenetic relevance of anaerobic androgen biodegradation in the environment

et al. 2016

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Steroid hormones, such as androgens, are common surface-water contaminants. However, literature on the ecophysiological relevance of steroid-degrading organisms in the environment, particularly in anoxic ecosystems, is extremely limited. We previously reported that Steroidobacter denitrificans anaerobically degrades androgens through the 2,3-seco pathway. In this study, the genome of Sdo. denitrificans was completely sequenced. Transcriptomic data revealed gene clusters that were distinctly expressed during anaerobic growth on testosterone. We isolated and characterized the bifunctional 1-testosterone hydratase/dehydrogenase, which is essential for anaerobic degradation of steroid A-ring. Because of apparent substrate preference of this molybdoenzyme, corresponding genes, along with the signature metabolites of the 2,3-seco pathway, were used as biomarkers to investigate androgen biodegradation in the largest sewage treatment plant in Taipei, Taiwan. Androgen metabolite analysis indicated that denitrifying bacteria in anoxic sewage use the 2,3-seco pathway to degrade androgens. Metagenomic analysis and PCR-based functional assays showed androgen degradation in anoxic sewage by Thauera spp. through the action of 1-testosterone hydratase/dehydrogenase. Our integrative 'omics' approach can be used for culture-independent investigations of the microbial degradation of structurally complex compounds where isotopelabeled substrates are not easily available.

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Site-directed mutagenesis of the Thauera aromatica strain T1 tutE tutFDGH gene cluster

Cited by 8 publications

References 38 publications

Anaerobic Catabolism of Aromatic Compounds: a Genetic and Genomic View

Anaerobic Catabolism of Aromatic Compounds: a Genetic and Genomic View

Substrate-bound Structures of Benzylsuccinate Synthase Reveal How Toluene Is Activated in Anaerobic Hydrocarbon Degradation

Integrated multi-omics analyses reveal the biochemical mechanisms and phylogenetic relevance of anaerobic androgen biodegradation in the environment

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