Catabolism of 4-hydroxybenzoate in proceeds through initial oxidative decarboxylation by a FAD-dependent 4-hydroxybenzoate 1-hydroxylase

Vanberkel, W; Eppink, M.H.M.; Middelhoven, W.J.; Vervoort, Jacques; Rietjens, Ivonne M. C. M.

doi:10.1016/0378-1097(94)90128-7

Cited by 23 publications

(30 citation statements)

References 20 publications

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“…With both dihydroxybenzene growth substrates, the enzyme constituted up to 5% of the total protein. The catalytic and hydrodynamic properties of this flavoenzyme were indistinguishable from those of the phenolic monooxygenase induced by growth on 4-hydroxybenzoate [28], confirming its general role in the catabolism of dihydroxybenzene compounds in C. parapsilosis (Fig. 1).…”

Section: R E S U L T S Induction Of Hydroquinone Hydroxylasementioning

confidence: 64%

“…Hydroquinone hydroxylase was purified to apparent homogeneity in three chromatographic steps by an improved procedure of the protocol reported previously [28]. All purification steps were performed at 4 8C.…”

Section: Enzyme Purificationmentioning

confidence: 99%

“…1,2,4-Trihydroxybenzene 1,2-dioxygenase from C. parapsilosis CBS604 was partially purified as reported earlier [28]. Phenol hydroxylase from T. cutaneum CBS2466 was purified, essentially as described [12].…”

Section: Enzyme Purificationmentioning

confidence: 99%

“…During the enzymatic conversion of dihydroxybenzenes, no product inhibition occurred as shown by kinetic analysis in the presence of partially purified 1,2,4-trihydroxybenzene dioxygenase. 19 F NMR product analysis with partially purified enzyme has revealed that hydroquinone hydroxylase from C. parapsilosis catalyzes the conversion of 2-fluoro-1,4-dihydroxybenzene to a mixture of 1,2,4-trihydroxybenzene, 1-fluoro-2,3,5-trihydroxybenzene, 1-fluoro-2,4,5-trihydroxybenzene, 1-fluoro-2,3,6-trihydroxybenzene [28]. Therefore, it was of interest to study the regiospecificity of hydroxylation of the purified enzyme with asymmetric monofluorinated phenols.…”

Section: Catalytic Propertiesmentioning

confidence: 99%

“…We recently established that catabolism of 4-hydroxybenzoate in C. parapsilosis proceeds through the successive formation of 1,4-dihydroxybenzene and 1,2,4-trihydroxybenzene ( Fig. 1) and that this rather unusual reaction pathway involves the action of two flavoenzymes [28,29]. We here describe the purification and characterization of the phenolic monooxygenase that catalyzes the second step of this pathway.…”

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confidence: 91%

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Purification and properties of hydroquinone hydroxylase, a FAD-dependent monooxygenase involved in the catabolism of 4-hydroxybenzoate in Candida parapsilosis CBS604

Eppink¹,

Cammaart²,

Wassenaar³

et al. 2000

Eur J Biochem

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The ascomycetous yeast Candida parapsilosis CBS604 catabolizes 4-hydroxybenzoate through the initial formation of hydroquinone (1,4-dihydroxybenzene). High levels of hydroquinone hydroxylase activity are induced when the yeast is grown on either 4-hydroxybenzoate, 2,4-dihydroxybenzoate, 1,3-dihydroxybenzene or 1,4-dihydroxybenzene as the sole carbon source. The monooxygenase constitutes up to 5% of the total amount of protein and is purified to apparent homogeneity in three chromatographic steps. Hydroquinone hydroxylase from C. parapsilosis is a homodimer of about 150 kDa with each 76-kDa subunit containing a tightly noncovalently bound FAD. The flavin prosthetic group is quantitatively resolved from the protein at neutral pH in the presence of chaotropic salts. The apoenzyme is dimeric and readily reconstituted with FAD.Hydroquinone hydroxylase from C. parapsilosis catalyzes the ortho-hydroxylation of a wide range of monocyclic phenols with the stoichiometric consumption of NADPH and oxygen. With most aromatic substrates, no uncoupling of hydroxylation occurs. Hydroxylation of monofluorinated phenols is highly regiospecific with a preference for C6 hydroxylation. Binding of phenol highly stimulates the rate of flavin reduction by NADPH. At pH 7.6, 25 8C, this step does not limit the rate of overall catalysis.During purification, hydroquinone hydroxylase is susceptible towards limited proteolysis. Proteolytic cleavage does not influence the enzyme dimeric nature but results in relatively stable protein fragments of 55, 43, 35 and 22 kDa. N-Terminal peptide sequence analysis revealed the presence of two nick sites and showed that hydroquinone hydroxylase from C. parapsilosis is structurally related to phenol hydroxylase from Trichosporon cutaneum. The implications of these findings for the catalytic mechanism of hydroquinone hydroxylase are discussed.

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Section: R E S U L T S Induction Of Hydroquinone Hydroxylasementioning

confidence: 64%

Section: Enzyme Purificationmentioning

confidence: 99%

Section: Enzyme Purificationmentioning

confidence: 99%

Section: Catalytic Propertiesmentioning

confidence: 99%

mentioning

confidence: 91%

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Purification and properties of hydroquinone hydroxylase, a FAD-dependent monooxygenase involved in the catabolism of 4-hydroxybenzoate in Candida parapsilosis CBS604

Eppink¹,

Cammaart²,

Wassenaar³

et al. 2000

Eur J Biochem

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Flavoenzyme-Catalyzed Oxygenations and Oxidations of Phenolic Compounds

Moonen

Fraaije

Rietjens

et al. 2002

Advanced Synthesis & Catalysis

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Flavin‐dependent monooxygenases and oxidases play an important role in the mineralization of phenolic compounds. Because of their exquisite regioselectivity and stereoselectivity, these enzymes are of interest for the biocatalytic production of fine chemicals and food ingredients. In our group, we have characterized several flavoenzymes that act on phenolic compounds, including 4‐hydroxybenzoate 3‐hydroxylase, 3‐hydroxyphenylacetate 6‐hydroxylase, 4‐hydroxybenzoate 1‐hydroxylase (decarboxylating), hydroquinone hydroxylase, 2‐hydroxybiphenyl 3‐monooxygenase, phenol hydroxylase, 4‐hydroxyacetophenone monooxygenase and vanillyl‐alcohol oxidase. The catalytic properties of these enzymes are reviewed here, together with insights obtained from site‐directed and random mutagenesis.

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Biocatalytic Potential of p‐Hydroxybenzoate Hydroxylase from Rhodococcus rhodnii 135 and Rhodococcus opacus 557

et al. 2004

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The biocatalytic potential of the NADHdependent p-hydroxybenzoate hydroxylases (PHBH) from Rhodococcus rhodnii 135 and Rhodococcus opacus 557 was investigated. Monofluorinated 4-hydroxybenzoates were efficiently hydroxylated, albeit at different rates. 2-Fluoro-4-hydroxybenzoate was a true substrate for PHBH from R. rhodnii 135 but a substrate inhibitor for PHBH from R. opacus 557. Monochlorinated 4-hydroxybenzoates also acted as PHBH substrates, but with these compounds strong uncoupling of hydroxylation (formation of hydrogen peroxide) occurred. PHBH from R. rhodnii 135 preferred the 5'-hydroxylation of 2-chloro-4-hydroxybenzoate but the enzyme from R. opacus 557 favored the formation of 2-chloro-3,4-dihydroxybenzoate. Conversely, PHBH from R. rhodnii 135 regioselectively hydroxylated 2-fluoro-4-hydroxybenzoate to 2-fluoro-3,4-dihydroxybenzoate whereas the enzyme from R. opacus 557 also produced significant amounts of 2-fluoro-4,5-dihydroxybenzoate. At high NADH/substrate ratio, both 2-fluorodihydroxybenzoate products were further converted to 2-fluoro-3,4,5-trihydroxybenzoate. PHBH from R. rhodnii 135 and R. opacus 557 preferred the 5'-hydroxylation of 3-chloro-4-hydroxybenzoate. However, conversion of 3-fluoro-4-hydroxybenzoate involved considerable dehalogenation affording nearly equal amounts of 3,4-dihydroxybenzoate and 5-fluoro-3,4-dihydroxybenzoate. At high NADH/substrate ratio, the latter compound was further converted to 3,4,5-trihydroxybenzoate. The results are discussed in relation to the properties of the NADPH-specific PHBH from Pseudomonas fluorescens.

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Catabolism of 4-hydroxybenzoate in proceeds through initial oxidative decarboxylation by a FAD-dependent 4-hydroxybenzoate 1-hydroxylase

Cited by 23 publications

References 20 publications

Purification and properties of hydroquinone hydroxylase, a FAD-dependent monooxygenase involved in the catabolism of 4-hydroxybenzoate in Candida parapsilosis CBS604

Purification and properties of hydroquinone hydroxylase, a FAD-dependent monooxygenase involved in the catabolism of 4-hydroxybenzoate in Candida parapsilosis CBS604

Flavoenzyme-Catalyzed Oxygenations and Oxidations of Phenolic Compounds

Biocatalytic Potential of p‐Hydroxybenzoate Hydroxylase from Rhodococcus rhodnii 135 and Rhodococcus opacus 557

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