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Literature summary for 1.14.13.24 extracted from
- Purification and characterization of the ncgl2923-encoded 3-hydroxybenzoate 6-hydroxylase from Corynebacterium glutamicum (2010), J. Basic Microbiol., 50, 599-604.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| gene ncgl2923, sequence determination, analysis, and comparisons, functional overexpression in of N-terminally His-tagged Ncgl2923 in Escherichia coli strain BL21(DE3), subcloning in Escherichia coli strain DH5alpha | Corynebacterium glutamicum |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.0534 | - |
3-hydroxybenzoate | recombinant enzyme, with NADH, pH 7.5, 25°C | Corynebacterium glutamicum |  |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 49000 | - |
3 * 49000, recombinant Ncgl2923, SDS-PAGE | Corynebacterium glutamicum |
| 125000 | - |
recombinant Ncgl2923, gel filtration | Corynebacterium glutamicum |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 3-hydroxybenzoate + NADH + H+ + O2 | Corynebacterium glutamicum | - |
2,5-dihydroxybenzoate + NAD+ + H2O | i.e. gentisate | ? | |
| 3-hydroxybenzoate + NADH + H+ + O2 | Corynebacterium glutamicum ATCC 13032 | - |
2,5-dihydroxybenzoate + NAD+ + H2O | i.e. gentisate | ? | |
| 3-hydroxybenzoate + NADPH + H+ + O2 | Corynebacterium glutamicum | - |
2,5-dihydroxybenzoate + NADP+ + H2O | i.e. gentisate | ? | |
| 3-hydroxybenzoate + NADPH + H+ + O2 | Corynebacterium glutamicum ATCC 13032 | - |
2,5-dihydroxybenzoate + NADP+ + H2O | i.e. gentisate | ? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Corynebacterium glutamicum | Q8NLB6 | gene ncgl2923 | - |
| Corynebacterium glutamicum ATCC 13032 | Q8NLB6 | gene ncgl2923 | - |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant N-terminally His-tagged Ncgl2923 from Escherichia coli strain BL21(DE3) by nickel affinity chromatography to homogeneity | Corynebacterium glutamicum |
Reaction
| Reaction | Comment | Organism | Reaction ID |
|---|---|---|---|
| 3-hydroxybenzoate + NADH + H+ + O2 = 2,5-dihydroxybenzoate + NAD+ + H2O | catalytic mechanism involving FAD, overview | Corynebacterium glutamicum |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 6.92 | - |
purified recombinant enzyme, with NADH, pH 7.5, 25°C | Corynebacterium glutamicum |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 3-hydroxybenzoate + NADH + H+ + O2 | - |
Corynebacterium glutamicum | 2,5-dihydroxybenzoate + NAD+ + H2O | i.e. gentisate | ? | |
| 3-hydroxybenzoate + NADH + H+ + O2 | - |
Corynebacterium glutamicum ATCC 13032 | 2,5-dihydroxybenzoate + NAD+ + H2O | i.e. gentisate | ? | |
| 3-hydroxybenzoate + NADPH + H+ + O2 | - |
Corynebacterium glutamicum | 2,5-dihydroxybenzoate + NADP+ + H2O | i.e. gentisate | ? | |
| 3-hydroxybenzoate + NADPH + H+ + O2 | - |
Corynebacterium glutamicum ATCC 13032 | 2,5-dihydroxybenzoate + NADP+ + H2O | i.e. gentisate | ? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| trimer | 3 * 49000, recombinant Ncgl2923, SDS-PAGE | Corynebacterium glutamicum |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| 3-hydroxybenzoate 6-hydroxylase | - |
Corynebacterium glutamicum |
| Ncgl2923 | - |
Corynebacterium glutamicum |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 25 | - |
assay at | Corynebacterium glutamicum |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7.5 | - |
- |
Corynebacterium glutamicum |
pH Range
| pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|
| 6 | 8 | high activity range | Corynebacterium glutamicum |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| FAD | 3fold activation at 0.015 mM. Sequence analysis of FAD/NADPH-binding domain of Ncgl2923, overview | Corynebacterium glutamicum | |
| NADH | dependent on, preferred cofactor | Corynebacterium glutamicum | |
| NADPH | less active than NADH | Corynebacterium glutamicum | |
General Information
| General Information | Comment | Organism |
|---|---|---|
| metabolism | strain ATCC 13032 metabolizes 3-hydroxybenzoate, the 3-hydroxybenzoate 6-hydroxylase is involved in the initial step of 3-hydroxybenzoate catabolismate via gentisate | Corynebacterium glutamicum |
| physiological function | the enzyme catalyzes the NADH-dependent conversion of 3-hydroxybenzoate to gentisate | Corynebacterium glutamicum |
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