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Literature summary for 5.99.1.4 extracted from
- 2-Hydroxychromene-2-carboxylate isomerase from bacteria that degrade naphthalenesulfonates (1993), Biodegradation, 4, 155-162.
No PubMed abstract available
Activating Compound
| Activating Compound | Comment | Organism | Structure |
|---|---|---|---|
| 2-mercaptoethanol | - |
Comamonas testosteroni |  |
| dithiothreitol | - |
Comamonas testosteroni | |
| glutathione | highest enzyme activity is found after preincubation of the enzyme with glutathione at alkaline pH-values. To determine enzyme activities during the purification procedure, it is necessary to reactivate the enzyme with glutathione | Comamonas testosteroni | |
| glutathione | no enzyme activity is found when the enzyme preparations have been preincubated without or with only 10 mM glutathione | Brevundimonas vesicularis | |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| additional information | no significant inhibition is observed after one day incubation with 50 mM EDTA | Brevundimonas vesicularis | |
| additional information | no significant inhibition is observed after one day incubation with 50 mM EDTA | Comamonas testosteroni |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.23 | - |
2-hydroxychromene-2-carboxylate | - |
Comamonas testosteroni | |
| 0.27 | - |
2-hydroxy-2H-chromene-2-carboxylate | - |
Brevundimonas vesicularis | |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| additional information | no increase of enzyme activity is found after (pre)incubation of the enzyme with ZnSO4, COCl2, FeCl2 or MnCl2 (each 2 mM) | Comamonas testosteroni | |
| additional information | no increase of enzyme activity is found after (pre)incubation of the enzyme with ZnSO4, COCl2, FeCl2 or MnCl2 (each 2 mM) | Brevundimonas vesicularis |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 24700 | - |
SDS-PAGE | Comamonas testosteroni |
| 24700 | - |
SDS-PAGE | Brevundimonas vesicularis |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| additional information | Comamonas testosteroni | enzyme degrades naphthalenesulfonates | ? | - |
? | |
| additional information | Brevundimonas vesicularis | enzyme degrades naphthalenesulfonates | ? | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Brevundimonas vesicularis | - |
BN6, DSM 6383 | - |
| Comamonas testosteroni | - |
A3, DSM 676 | - |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| at room temperature by use of a fast-protein liquid chromatography system | Comamonas testosteroni |
| partially purified by anion-exchange chromatography | Brevundimonas vesicularis |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 0.005 | - |
spectrophotometrically | Comamonas testosteroni |
| 0.005 | - |
spectrophotometrically | Brevundimonas vesicularis |
| 0.14 | - |
for 2-hydroxychromene-2-carboxylate as substrate | Brevundimonas vesicularis |
| 0.2 | - |
enzyme activity increases when the cell extract is incubated for 15 min with glutathione (5 mM) prior to the assay. The addition of glutathione (0.2 mM) directly to the assay does not increase enzyme activity | Comamonas testosteroni |
| 0.2 | - |
enzyme activity increases when the cell extract is incubated for 15 min with glutathione (5 mM) prior to the assay. The addition of glutathione (0.2 mM) directly to the assay does not increase enzyme activity | Brevundimonas vesicularis |
| 0.27 | - |
for 2-hydroxychromene-2-carboxylate as substrate | Comamonas testosteroni |
| 55.4 | - |
last purification step | Comamonas testosteroni |
Storage Stability
| Storage Stability | Organism |
|---|---|
| 4°C, purifed enzyme, Tris HCl, pH 7.5, 100 mM NaCl, 1 month 52% loss of activity | Comamonas testosteroni |
| 4°C, purifed enzyme, Tris HCl, pH 7.5, 100 mM NaCl, 1 month 52% loss of activity | Brevundimonas vesicularis |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 2-hydroxy-2H-chromene-2-carboxylate | - |
Comamonas testosteroni | (3E)-4-(2-hydroxyphenyl)-2-oxobut-3-enoate | - |
? | |
| 2-hydroxy-2H-chromene-2-carboxylate | - |
Brevundimonas vesicularis | (3E)-4-(2-hydroxyphenyl)-2-oxobut-3-enoate | - |
? | |
| 2-hydroxybenzo[g]chromene-2-carboxylate | more unstable than 2-hydroxychromene-2-carboxylate | Comamonas testosteroni | ? | - |
? | |
| 2-hydroxybenzo[g]chromene-2-carboxylate | more unstable than 2-hydroxychromene-2-carboxylate | Brevundimonas vesicularis | ? | - |
? | |
| additional information | enzyme degrades naphthalenesulfonates | Comamonas testosteroni | ? | - |
? | |
| additional information | enzyme degrades naphthalenesulfonates | Brevundimonas vesicularis | ? | - |
? | |
| additional information | 2,5-dihydroxychromene-2-carboxylate is no substrate | Comamonas testosteroni | ? | - |
? | |
| additional information | 2,5-dihydroxychromene-2-carboxylate is no substrate | Brevundimonas vesicularis | ? | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| monomer | 1 * 24700, SDS-PAGE | Comamonas testosteroni |
| monomer | 1 * 24700, SDS-PAGE | Brevundimonas vesicularis |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| 2-hydroxychromene-2-carboxylate isomerase | - |
Comamonas testosteroni |
| 2-hydroxychromene-2-carboxylate isomerase | - |
Brevundimonas vesicularis |
| 2HCCAI | - |
Comamonas testosteroni |
| 2HCCAI | - |
Brevundimonas vesicularis |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 9 | - |
in glycine-NaOH-buffer, highest enzyme activity is found after preincubation of the enzyme with glutathione at alkaline pH-values | Comamonas testosteroni |
| 9.5 | - |
in glycine-NaOH-buffer | Brevundimonas vesicularis |
| 10 | - |
assay at | Comamonas testosteroni |
pH Range
| pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|
| 8 | 9.5 | at pH 9.0 50% and at pH 8.0 only 5% of the maximal activity is found, respectively | Brevundimonas vesicularis |
| 8 | 9.3 | effect of glutathione is optimal between pH 8 and pH 9.3. In contrast, no activation is observed when the enzyme is incubated with glutathione at pH 5.0. Activities higher than 70% of this optimal activity are found at pH-values in the range 8.0-9.5 in glycine-NaOH- or Tris/HC1-buffer. At pH 7 about 50% and at pH 5.5 10% of the optimal activity is observed | Comamonas testosteroni |
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