Benzoyl‐Coenzyme‐A 3‐Monooxygenase, a Flavin‐Dependent Hydroxylase

Niemetz, Ruth; Altenschmidt, Uwe; Brucker, Susanne; Fuchs, Georg

doi:10.1111/j.1432-1033.1995.tb20372.x

Cited by 22 publications

(22 citation statements)

References 38 publications

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“…The assay mixture (600 l) contained 100 mM Tris-HCl (pH 8.5), 2 mM dithiothreitol, 5 mM MgCl 2 , 1 mM ATP, 0.4 mM CoA, 1 mM aromatic acid, and 6 l of crude cell extract. Benzoyl-CoA was measured at 290 nm (ε ϭ 3.9 mM Ϫ1 cm Ϫ1 ), 2-hydroxybenzoyl-CoA was measured at 320 nm (ε ϭ 5.9 mM Ϫ1 cm Ϫ1 ), 3-hydroxybenzoyl-CoA was measured at 310 nm (ε ϭ 3.3 mM Ϫ1 cm Ϫ1 ), 4-hydroxybenzoyl-CoA was measured at 300 nm (ε ϭ 10.55 mM Ϫ1 cm Ϫ1 ), and 2-aminobenzoyl-CoA was measured at 365 nm (ε ϭ 5.5 mM Ϫ1 cm Ϫ1 ) (48,60). For the coupled enzyme assay, AMP formation was monitored by coupling the CoA ligase reaction to the myokinase, pyruvate kinase, and lactate dehydrogenase system and by spectrophotometrically measuring the rate of NADH oxidation at 365 nm (60).…”

Section: Methodsmentioning

confidence: 99%

“…Benzoate-CoA ligase activity was determined at 30°C with crude cell extracts through a direct spectrophotometric assay or via a coupled enzyme assay. For the direct assay, product formation (CoA derivatives) was monitored spectrophotometrically as described previously (48). The assay mixture (600 l) contained 100 mM Tris-HCl (pH 8.5), 2 mM dithiothreitol, 5 mM MgCl 2 , 1 mM ATP, 0.4 mM CoA, 1 mM aromatic acid, and 6 l of crude cell extract.…”

Section: Methodsmentioning

confidence: 99%

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ThebzdGene Cluster, Coding for Anaerobic Benzoate Catabolism, inAzoarcussp. Strain CIB

et al. 2004

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We report here that the bzd genes for anaerobic benzoate degradation in Azoarcus sp. strain CIB are organized as two transcriptional units, i.e., a benzoate-inducible catabolic operon, bzdNOPQMSTUVWXYZA, and a gene, bzdR, encoding a putative transcriptional regulator. The last gene of the catabolic operon, bzdA, has been expressed in Escherichia coli and encodes the benzoate-coenzyme A (CoA) ligase that catalyzes the first step in the benzoate degradation pathway. The BzdA enzyme is able to activate a wider range of aromatic compounds than that reported for other previously characterized benzoate-CoA ligases. The reduction of benzoyl-CoA to a nonaromatic cyclic intermediate is carried out by a benzoyl-CoA reductase (bzdNOPQ gene products) detected in Azoarcus sp. strain CIB extracts. The bzdW, bzdX, and bzdY gene products show significant similarity to the hydratase, dehydrogenase, and ring-cleavage hydrolase that act sequentially on the product of the benzoyl-CoA reductase in the benzoate catabolic pathway of Thauera aromatica. Benzoate-CoA ligase assays and transcriptional analyses based on lacZ-reporter fusions revealed that benzoate degradation in Azoarcus sp. strain CIB is subject to carbon catabolite repression by some organic acids, indicating the existence of a physiological control that connects the expression of the bzd genes to the metabolic status of the cell.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

ThebzdGene Cluster, Coding for Anaerobic Benzoate Catabolism, inAzoarcussp. Strain CIB

et al. 2004

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“…Moreover, some dehalogenation mechanisms of aromatic compounds also involve CoA thioester formation in aerobiosis (81). Although the rationale for utilizing such hybrid pathways, i.e., aerobic catabolic pathways endowed with typical features of an anaerobic catabolism, is not known, it has been suggested that they could represent a strategy of the microorganisms to cope with fluctuations of oxygen supply (212). In this sense, the existence of a hybrid pathway for the catabolism of PA in E. coli could reflect the facultative anaerobic character of this bacterium.…”

Section: Pa Aerobic Hybrid Pathwaymentioning

confidence: 99%

Biodegradation of Aromatic Compounds by Escherichia coli

Dı́az

Ferrández

Prieto

et al. 2001

Microbiol Mol Biol Rev

322

235

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Although Escherichia coli has long been recognized as the best-understood living organism, little was known about its abilities to use aromatic compounds as sole carbon and energy sources. This review gives an extensive overview of the current knowledge of the catabolism of aromatic compounds by E. coli. After giving a general overview of the aromatic compounds that E. coli strains encounter and mineralize in the different habitats that they colonize, we provide an up-to-date status report on the genes and proteins involved in the catabolism of such compounds, namely, several aromatic acids (phenylacetic acid, 3- and 4-hydroxyphenylacetic acid, phenylpropionic acid, 3-hydroxyphenylpropionic acid, and 3-hydroxycinnamic acid) and amines (phenylethylamine, tyramine, and dopamine). Other enzymatic activities acting on aromatic compounds in E. coli are also reviewed and evaluated. The review also reflects the present impact of genomic research and how the analysis of the whole E. coli genome reveals novel aromatic catabolic functions. Moreover, evolutionary considerations derived from sequence comparisons between the aromatic catabolic clusters of E. coli and homologous clusters from an increasing number of bacteria are also discussed. The recent progress in the understanding of the fundamentals that govern the degradation of aromatic compounds in E. coli makes this bacterium a very useful model system to decipher biochemical, genetic, evolutionary, and ecological aspects of the catabolism of such compounds. In the last part of the review, we discuss strategies and concepts to metabolically engineer E. coli to suit specific needs for biodegradation and biotransformation of aromatics and we provide several examples based on selected studies. Finally, conclusions derived from this review may serve as a lead for future research and applications

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“…strain B4 suggested gentisyl-CoA as an intermediate and thus a possible role for gentisyl-CoA thioesterase (11). Also, the first steps of a novel benzoate aerobic degradation pathway observed in a denitrifying bacterium may involve transformation of benzoate to benzoyl-CoA, benzoyl-CoA to 3-hydroxybenzoyl-CoA, and 3-hydroxybenzoyl-CoA to gentisyl-CoA (1,19). Finally, it has been postulated that gentisyl-CoA is formed from 4-hydroxbenzoyl-CoA in halophilic Archaea via the action of a novel monooxygenase that catalyzes intramolecular thioester group migration (i.e., the NIH shift) (6).…”

Section: Chemicalsmentioning

confidence: 99%

The BH1999 Protein of Bacillus halodurans C-125 Is Gentisyl-Coenzyme A Thioesterase

et al. 2004

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In this study, we have shown that recombinant BH1999 from Bacillus halodurans catalyzes the hydrolysis of gentisyl coenzyme A (CoA) (2,5-dihydroxybenzoyl-coenzyme A) at physiological pH with a k cat /K m of 1.6 ؋ 10 6 M ؊1 s ؊1 and the hydrolysis of 3-hydroxybenzoyl-CoA with a k cat /K m of 3.0 ؋ 10 5 M ؊1 s ؊1 . All other acyl-CoA thioesters tested had low or no substrate activity. The BH1999 gene is juxtaposed with a gene cluster that contains genes believed to function in gentisate oxidative degradation. It is hypothesized that BH1999 functions as a gentisyl-CoA thioesterase. Gentisyl-CoA thioesterase shares the backbone fold and the use of an active site aspartate residue to mediate catalysis with the 4-hydroxybenzoyl-CoA thioesterase of the hotdog fold enzyme superfamily. A comparative study of these two enzymes showed that they differ greatly in the rate contribution made by the catalytic aspartate, in the pH dependence of catalysis, and in substrate specificity.The pathways leading to the degradation of environmental aromatic compounds in bacteria and fungi form the foundation for the bioremediation of industrial waste products. Studies have shown that a wide variety of aromatic compounds are degraded through pathways that converge at catechol or protocatechuate (12). These intermediates, in turn, undergo oxidative ring opening, followed by mineralization via the ␤-ketoadipate pathway (12). Gentisate (2,5-dihydroxybenzoate) is another common intermediate that has been implicated in the degradation pathways of 3-hydroxybenzoate (9), xylenol (20), salicylate (21), 3,6-dichloro-2-methoxybenzoate (28), and naphthalene (8). Gentisate undergoes oxidative ring cleavage (catalyzed by gentisate 1,2-dioxygenase) to maleylpyruvate. Maleylpyruvate is either cleaved to pyruvate and maleate by the enzyme maleylpyruvate hydrolase or converted to fumarylpyruvate with maleylpyruvate isomerase. The fumarylpyruvate is then cleaved to fumarate and pyruvate by fumarylpyruvate hydrolase (2,14,20,22).In Bacillus halodurans C-125 (26), gentisate degradation via oxidation to maleylpyruvate and cleavage to pyruvate is evidenced by the gene cluster represented in Fig. 1. The gene cluster contains two genes encoding homologs to gentisate 1,2-dioxygenase (BH2002 and BH2004, which have 32 to 37% sequence identity to the previously characterized gentisate 1,2-dioxygenase from Sphingomonas sp. strain RW5 [28]) and Ralstonia sp. strain U2 (30). In addition, the neighboring gene BH2005 encodes a homolog of fumarylacetoacetate hydrolase, an enzyme that catalyzes hydrolytic cleavage in the close analog fumarylacetoacetate (17). The presence of an isomerase homologue (BH2000) may indicate the formation of fumarylpyruvate. The upstream gene BH1999 encodes a small protein which is homologous to the 4-hydroxybenzoyl-coenzyme A (CoA) thioesterases from Pseudomonas sp. strain DJ12 (35% identity in amino acid sequence) (5) and strain CBS3 (31% identity in amino acid sequence) (3). It seemed plausible that BH1999 is a gentisyl-CoA thioesterase.To t...

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Benzoyl‐Coenzyme‐A 3‐Monooxygenase, a Flavin‐Dependent Hydroxylase

Cited by 22 publications

References 38 publications

ThebzdGene Cluster, Coding for Anaerobic Benzoate Catabolism, inAzoarcussp. Strain CIB

ThebzdGene Cluster, Coding for Anaerobic Benzoate Catabolism, inAzoarcussp. Strain CIB

Biodegradation of Aromatic Compounds by Escherichia coli

The BH1999 Protein of Bacillus halodurans C-125 Is Gentisyl-Coenzyme A Thioesterase

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