Cloned (Comment) | Organism |
---|---|
gene mnx1, sequence comparisons and phylogenetic analysis and tree, recombinant expression of C-terminally His-tagged enzyme in Escherichia coli strain TOP10 | Candida parapsilosis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.01 | - |
4-hydroxybenzoate | recombinant enzyme, pH 7.6, 25°C | Candida parapsilosis |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-hydroxybenzoate + NADH + 2 H+ + O2 | Candida parapsilosis | - |
hydroquinone + NAD+ + H2O + CO2 | - |
? | |
4-hydroxybenzoate + NADH + 2 H+ + O2 | Candida parapsilosis CBS604 | - |
hydroquinone + NAD+ + H2O + CO2 | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Candida parapsilosis | - |
- |
- |
Candida parapsilosis CBS604 | - |
- |
- |
Posttranslational Modification | Comment | Organism |
---|---|---|
additional information | 4HB1H does not contain a lipid cofactor | Candida parapsilosis |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged enzyme from Escherichia coli strain TOP10 by nickel affinity chromatography, dialysis, ion exchange chromatography, and gel filtration | Candida parapsilosis |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
4-hydroxybenzoate + NAD(P)H + H+ + O2 = hydroquinone + NAD(P)+ + H2O + CO2 | the conversion of 4-HB involves an ipso-attack of the electrophilic flavin C4a-hydroperoxide at the C1-atom of the activated substrate, resulting in a tetrahedral benzoquinone species and subsequent release of the carboxyl side chain. It cannot be excluded that His230 activates 4-HB in 4HB1H for ipso-attack by the flavin C4a-hydroperoxide. Furthermore, it can also not be ruled out that the substrate of 4HB1H binds in a flipped orientation with the 4-hydroxyl moiety pointing towards the side chains of Tyr116 and His118 | Candida parapsilosis |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
14 | - |
purified recombinant enzyme, pH 7.6, 25°C, substrates 4-hydroxybenzoate and NADH | Candida parapsilosis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-hydroxybenzoate + NADH + 2 H+ + O2 | - |
Candida parapsilosis | hydroquinone + NAD+ + H2O + CO2 | - |
? | |
4-hydroxybenzoate + NADH + 2 H+ + O2 | - |
Candida parapsilosis CBS604 | hydroquinone + NAD+ + H2O + CO2 | - |
? | |
additional information | 4HB1H catalyzes an oxidative decarboxylation reaction. 4HB1H is active with a wide range of 4-hydroxybenzoate (4-HB) derivatives. 4HB1H can also be induced by protocatechuate (PCA) and convert this compound directly into the central intermediate HHQ | Candida parapsilosis | ? | - |
- |
|
additional information | 4HB1H catalyzes an oxidative decarboxylation reaction. 4HB1H is active with a wide range of 4-hydroxybenzoate (4-HB) derivatives. 4HB1H can also be induced by protocatechuate (PCA) and convert this compound directly into the central intermediate HHQ | Candida parapsilosis CBS604 | ? | - |
- |
|
protocatechuate + NADH + 2 H+ + O2 | - |
Candida parapsilosis | hydroquinone + NAD+ + H2O + CO2 | - |
? | |
protocatechuate + NADH + 2 H+ + O2 | - |
Candida parapsilosis CBS604 | hydroquinone + NAD+ + H2O + CO2 | - |
? | |
vanillate + NADH + 2 H+ + O2 | - |
Candida parapsilosis | 3-methoxy-1,4-dihydroxybenzene + NAD+ + H2O + CO2 | - |
? | |
vanillate + NADH + 2 H+ + O2 | - |
Candida parapsilosis CBS604 | 3-methoxy-1,4-dihydroxybenzene + NAD+ + H2O + CO2 | - |
? |
Subunits | Comment | Organism |
---|---|---|
monomer | - |
Candida parapsilosis |
Synonyms | Comment | Organism |
---|---|---|
4-hydroxybenzoate 1-hydroxylase (decarboxylating) | - |
Candida parapsilosis |
4HB1H | - |
Candida parapsilosis |
Cp_4HB1H | - |
Candida parapsilosis |
FAD-dependent 4-hydroxybenzoate hydroxylase | - |
Candida parapsilosis |
PHBH | - |
Candida parapsilosis |
V1H | - |
Candida parapsilosis |
vanillate 1-hydroxylase | - |
Candida parapsilosis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
- |
Candida parapsilosis |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
10 | - |
4-hydroxybenzoate | recombinant enzyme, pH 7.6, 25°C | Candida parapsilosis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.6 | - |
- |
Candida parapsilosis |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FAD | addition of FAD is essential to achieve optimal turnover | Candida parapsilosis | |
additional information | 4HB1H does not contain a lipid cofactor | Candida parapsilosis | |
NADH | highly preferred cofactor | Candida parapsilosis |
General Information | Comment | Organism |
---|---|---|
evolution | phylogenetic analysis of FAD-dependent 4-hydroxybenzoate hydroxylases and 3-hydroxybenzoate 4-hydroxylase, phylogenetic analysis and tree, overview. Enzyme 4HB1H is only present in a small group of Saccharomycetes. Many FAD-binding proteins with unknown function in pezizomycetes and agaricomycetes have structural features in common with 4HB1H, but lack its C-terminal tail. 4HB1H belongs to a subgroup of structurally related group A FPMOs that catalyze para-hydroxylation reactions | Candida parapsilosis |
metabolism | the enzyme is important in lignin degradation. The conversion of 4-hydroxybenzoate (4-HB) involves an ipso-attack of the electrophilic flavin C4a-hydroperoxide at the C1-atom of the activated substrate, resulting in a tetrahedral benzoquinone species and subsequent release of the carboxyl side chain. The second step of the catabolism of 4-HB in Candida parapsilosis catalyzed by the homodimeric hydroquinone hydroxylase, HQH | Candida parapsilosis |
additional information | structure comparisons and protein homology modelling, generation of a model of Candida parapsilosis Cp_HQH and models for Cp_4HB1H, Fusarium oxysporum Fo_4HB1H-like, and Boreostereum vibrans Bv_VibMO1 using Cutaneotrichosporon cutaneum Tc_PHHY (PDB ID 1pn0), Rhodococcus jostii Rj_3HB6H (PDB ID 4bk1) and Pseudomonas putida Pp_SALH (PDB ID 5evy) as template structures, overview | Candida parapsilosis |
physiological function | 4-hydroxybenzoate 1-hydroxylase (4HB1H) from yeast catalyzes an oxidative decarboxylation reaction and is structurally similar to 3-hydroxybenzoate 6-hydroxylase (3HB6H), salicylate hydroxylase (SALH) and 6-hydroxynicotinate 3-monooxygenase (6HNMO). Genome mining suggests that the 4HB1H activity is widespread in the fungal kingdom and might be responsible for the oxidative decarboxylation of vanillate, an import intermediate in lignin degradation | Candida parapsilosis |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1000 | - |
4-hydroxybenzoate | recombinant enzyme, pH 7.6, 25°C | Candida parapsilosis |