3-Hydroxybenzoate 6-hydroxylase from Pseudomonas aeruginosa

Groseclose, Edye E.; Ribbons, Douglas W.

doi:10.1016/0006-291x(73)91228-x

Cited by 43 publications

(25 citation statements)

References 15 publications

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“…Data on comparative sizes of phenol hydroxylase from bacterial sources are lacking since the enzyme has not been completely purified from procaryotes; however, the 80-kilodalton relative molecular mass determined for the P. pickettii PKO1 phenol hydroxylase in this study is comparable to the 74-kilodalton monomer molecular mass for phenol hydroxylase from the yeast T. cutaneum (28) and is in the same size range as that found for several other bacterial aromatic flavoprotein hydroxylases (14,17,27,39). Plasmidencoded dichlorophenol hydroxylases have recently been purified from several bacterial strains that are capable of degrading chlorophenoxy herbicides (4,25), but these enzymes show no activity toward phenol.…”

Section: Pseudomonas Pickettii Phenol Hydroxylase 4629supporting

confidence: 51%

Molecular cloning, characterization, and regulation of a Pseudomonas pickettii PKO1 gene encoding phenol hydroxylase and expression of the gene in Pseudomonas aeruginosa PAO1c

Kukor

Olsen

1990

J Bacteriol

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A 26-kilobase BamlI restriction endonuclease DNA fragment was cloned from Pseudomonas pickeUii PKO1, a strain isolated from a soil microcosm that had been amended with benzene, toluene, and xylene. This DNA fragment, cloned into vector plasmid pRO1727 and designated pRO1957, allowed Pseudomonas aeruginosa PAO1c to grow on phenol as the sole source of carbon. Physical and functional restriction endonudease maps have been derived for the cloned DNA fragment. Two DNA fragments carried in trans and derived from subclones of pRO1957 show phenol hydroxylase activity in cell extracts of P. aeruginosa. Deletion and subcloning analyses of these fragments indicated that the gene encoding phenol hydroxylase is positively regulated. Phenol and m-cresol were shown to be inducers of the enzyme. o-Cresol and p-cresol did not induce enzymatic activity but could be metabolized by cells that had been previously exposed to phenol or m-cresol; moreover, the enzyme exhibited a rather broad substrate specificity and was sensitive to thiol-inhibiting reagents. A novel polypeptide with an estimated molecular mass of 80,000 daltons was detected in extracts of phenol-induced cells of P. aeruginosa carrying plasmid pRO1959.

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Section: Pseudomonas Pickettii Phenol Hydroxylase 4629supporting

confidence: 51%

Molecular cloning, characterization, and regulation of a Pseudomonas pickettii PKO1 gene encoding phenol hydroxylase and expression of the gene in Pseudomonas aeruginosa PAO1c

Kukor

Olsen

1990

J Bacteriol

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“…XlnD, along with other characterized 3-hydroxybenzoate 6-hydroxylases, was able to utilize both NADH and NADPH as cofactors. With 3-hydroxybenzoate as the substrate, XlnD has a preference for NADH over NADPH, a trait that it shares with the 3-hydroxybenzoate 6-hydroxylases from P. aeruginosa (6) and Micrococcus sp. (22).…”

Section: Discussionmentioning

confidence: 99%

“…The genetics and catalytic mechanism of hydroxylases that catalyze hydroxylation at a position ortho to an existing hydroxyl group of the aromatic ring has been widely characterized for 4-hydroxybenzoate hydroxylase (1,4,5,19,27). However, very little is known about the genetics and biochemistry of hydroxylases that catalyze the hydroxylation of the aromatic ring at a position para to an existing hydroxyl group, with 3-hydroxybenzoate 6-hydroxylase having been purified and characterized thus far only from Pseudomonas aeruginosa (6), Micrococcus sp. (22).…”

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confidence: 99%

Molecular and Biochemical Characterization of the xlnD -Encoded 3-Hydroxybenzoate 6-Hydroxylase Involved in the Degradation of 2,5-Xylenol via the Gentisate Pathway in Pseudomonas alcaligenes NCIMB 9867

et al. 2005

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The xlnD gene from Pseudomonas alcaligenes NCIMB 9867 (strain P25X) was shown to encode 3-hydroxybenzoate 6-hydroxylase I, the enzyme that catalyzes the NADH-dependent conversion of 3-hydroxybenzoate to gentisate. Active recombinant XlnD was purified as a hexahistidine fusion protein from Escherichia coli, had an estimated molecular mass of 130 kDa, and is probably a trimeric protein with a subunit mass of 43 kDa. This is in contrast to the monomeric nature of the few 3-hydroxybenzoate 6-hydroxylases that have been characterized thus far. Like other 3-hydroxybenzoate 6-hydroxylases, XlnD could utilize either NADH or NADPH as the electron donor. P25X harbors a second 3-hydroxybenzoate 6-hydroxylase II that was strictly inducible by specific aromatic substrates. However, the degradation of 2,5-xylenol and 3,5-xylenol in strain P25X was found to be dependent on the xlnD-encoded 6-hydroxylase I and not the second, strictly inducible 6-hydroxylase II.

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“…Wheelis et al (34) suggested an aerobic benzoate metabolism via 3-hydroxybenzoate and gentisate for which a benzoate 3-hydroxylase has to be postulated. This enzyme has never been demonstrated, in contrast to 3-hydroxybenzoate 6-hydroxylase (19). Also, in cell extracts of Pseudomonas strain KB 740, a benzoate 3-hydroxylase activity could not be detected.…”

mentioning

confidence: 83%

“…Extracts of Pseudomonas strain KB 740 catalyzed the hydroxylation of both 3-hydroxybenzoate and 3-hydroxybenzoyl-CoA with NAD(P)H at similar rates but with higher affinity for 3-hydroxybenzoyl-CoA, and in both cases gentisate was the product; we do not know yet whether these two reactions are catalyzed by one or more enzymes (3a). An FADdependent 3-hydroxybenzoate 6-hydroxylase was found in Pseudomonas aeruginosa by Groseclose and Ribbons (19), and Wheelis et al (34) described an NADPH-dependent enzyme in Pseudomonas testosteroni. The following ring cleavage reaction is postulated to be performed by gentisate dioxygenase.…”

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confidence: 97%

New aerobic benzoate oxidation pathway via benzoyl-coenzyme A and 3-hydroxybenzoyl-coenzyme A in a denitrifying Pseudomonas sp

et al. 1993

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A denitrifying Pseudomonas sp. is able to oxidize aromatic compounds compounds completely to CO2, both aerobically and anaerobically. It is shown that benzoate is aerobically oxidized by a new degradation pathway via benzoyl-coenzyme A (CoA) and 3-hydroxybenzoyl-CoA. The organism grew aerobically with benzoate, 3-hydroxybenzoate, and gentisate; catechol, 2-hydroxybenzoate, and protocatechuate were not used, and 4-hydroxybenzoate was a poor substrate. Mutants were obtained which were not able to utilize benzoate as the sole carbon source aerobically but still used 3-hydroxybenzoate or gentisate. Simultaneous adaptation experiments with whole cells seemingly suggested a sequential induction of enzymes of a benzoate oxidation pathway via 3-hydroxybenzoate and gentisate. Cells grown aerobically with benzoate contained a benzoate-CoA ligase (AMP forming) (0.1 mumol min-1 mg-1) which converted benzoate but not 3-hydroxybenzoate into its CoA thioester. The enzyme of 130 kDa composed of two identical subunits of 56 kDa was purified and characterized. Cells grown aerobically with 3-hydroxybenzoate contained a similarly active CoA ligase for 3-hydroxybenzoate, 3-hydroxybenzoate-CoA ligase (AMP forming). Extracts from cells grown aerobically with benzoate catalyzed a benzoyl-CoA- and flavin adenine dinucleotide-dependent oxidation of NADPH with a specific activity of at least 25 nmol NADPH oxidized min-1 mg of protein-1; NADH and benzoate were not used. This new enzyme, benzoyl-CoA 3-monooxygenase, was specifically induced during aerobic growth with benzoate and converted [U-14C]benzoyl-CoA stoichiometrically to [14C]3-hydroxybenzoyl-CoA.

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3-Hydroxybenzoate 6-hydroxylase from Pseudomonas aeruginosa

Cited by 43 publications

References 15 publications

Molecular cloning, characterization, and regulation of a Pseudomonas pickettii PKO1 gene encoding phenol hydroxylase and expression of the gene in Pseudomonas aeruginosa PAO1c

Molecular cloning, characterization, and regulation of a Pseudomonas pickettii PKO1 gene encoding phenol hydroxylase and expression of the gene in Pseudomonas aeruginosa PAO1c

Molecular and Biochemical Characterization of the xlnD -Encoded 3-Hydroxybenzoate 6-Hydroxylase Involved in the Degradation of 2,5-Xylenol via the Gentisate Pathway in Pseudomonas alcaligenes NCIMB 9867

New aerobic benzoate oxidation pathway via benzoyl-coenzyme A and 3-hydroxybenzoyl-coenzyme A in a denitrifying Pseudomonas sp

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