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Literature summary for 4.2.1.130 extracted from
- Glyoxalase III from Escherichia coli: a single novel enzyme for the conversion of methylglyoxal into D-lactate without reduced glutathione (1995), Biochem. J., 305, 999-1003.
General Stability
| General Stability | Organism |
|---|---|
| DTT and either sucrose or mannitol are required for stability | Escherichia coli |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| 4-hydroxymercuribenzoate | inactivates the enzyme, reversible by dithiothreitol and other thiol-group-containing compounds | Escherichia coli |  |
| 5,5'-dithiobis-(2-nitrobenzoate) | 0.4 mM, 15 min, complete inactivation. 5 mM DTT restores activity almost completely | Escherichia coli | |
| Borate | boric acid/borate buffer is moderately inhibitory | Escherichia coli | |
| DTNB | - |
Escherichia coli | |
| additional information | glutathione analogues, which are inhibitors of glyoxalase I, do not inhibit glyoxalase III, but the enzyme is sensitive to thiol-blocking reagents; S-methylglutathione and S-octylglutathione, even at concentrations of 20 mM and 0.25 mM respectively, have no inhibitory effect on Escherichia coli glyoxalase III | Escherichia coli | |
| N-ethylmaleimide | 0.4 mM, 15 min, complete inactivation. 5 mM DTT restores activity almost completely | Escherichia coli | |
| NEM | - |
Escherichia coli | |
| p-hydroxymercuribenzoate | 0.4 mM, 15 min, complete inactivation. 5 mM DTT restores activity almost completely | Escherichia coli | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 40000 | - |
2 * 40000, SDS-PAGE | Escherichia coli |
| 41000 | - |
- |
Escherichia coli |
| 82000 | - |
gel filtration | Escherichia coli |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 2-oxopropanal + H2O | Escherichia coli | - |
(R)-lactate | - |
ir | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Escherichia coli | - |
- |
- |
| Escherichia coli | P31658 | - |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
- |
Escherichia coli |
| native enzyme 725fold by ammonium sulfate fractionation, heat treatment, gel filtration, anion exchange chromatography, and affinity chromatography on a 4-hydroxymercuribenzoate-bound resin | Escherichia coli |
Source Tissue
| Source Tissue | Comment | Organism | Textmining |
|---|---|---|---|
| additional information | glyoxalase III activity is present in all three media in almost equal amounts ranging from 0.033 to 0.054 unit/mg of protein | Escherichia coli | - |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 0.033 | 0.054 | crude enzyme extracts, pH 8.0, 37°C | Escherichia coli |
| 37 | - |
pH 8.0, 37°C | Escherichia coli |
Storage Stability
| Storage Stability | Organism |
|---|---|
| -15°C, 1 mM DTT and 10% sucrose, almost completely stable | Escherichia coli |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 2-oxopropanal + H2O | - |
Escherichia coli | (R)-lactate | - |
ir | |
| 2-oxopropanal + H2O | stereospcific recation | Escherichia coli | (R)-lactate | - |
ir | |
| methylglyoxal + H2O | glyoxalase III does not catalyse the reverse reaction. Glyoxalase III cannot catalyse either the formation or the breakdown of S-D-lactoylglutathione | Escherichia coli | (R)-lactate | - |
ir | |
| additional information | purified glyoxalase III can catalyse the conversion of methylglyoxal into S-D-lactoylglutathione in the presence of GSH or S-D-lactoylglutathione into D-lactate, cf. EC 4.4.1.5 | Escherichia coli | ? | - |
? | |
| phenylglyoxal + H2O | - |
Escherichia coli | ? | - |
ir | |
| phenylglyoxal + H2O | the rate with phenylglyoxal is about 15% of that with methylglyoxal | Escherichia coli | ? | - |
ir | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| dimer | 2 * 40000, SDS-PAGE | Escherichia coli |
| dimer | 2 * 41000, about, SDS-PAGE | Escherichia coli |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| glyoxalase III | - |
Escherichia coli |
| glyoxylase III | - |
Escherichia coli |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 37 | - |
assay at | Escherichia coli |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 6 | 8 | no sharp optimum | Escherichia coli |
| 8 | - |
assay at | Escherichia coli |
| 8 | - |
glyoxalase III is active over a wide range of pH with no sharp pH optimum | Escherichia coli |
pH Range
| pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|
| 5 | 10 | pH 5.0: sharp decrease in reaction rate below, pH 10.0: activity 20-25% lower than the rate at physiological pH | Escherichia coli |
| 5 | 10 | glyoxalase III is active over a wide range of pH with no sharp pH optimum, sharp decrease in reaction rate occurs below pH 5.0, 20.25% of maximal activity at pH 10.0 | Escherichia coli |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| additional information | no requirement for glutathione, GSH | Escherichia coli |
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