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Literature summary for 4.2.1.53 extracted from
- Biochemical characterization and FAD-binding analysis of oleate hydratase from Macrococcus caseolyticus (2012), Biochimie, 94, 907-915.
Activating Compound
| Activating Compound | Comment | Organism | Structure |
|---|---|---|---|
| ethanol | maximum activity in the presence of 2% (v/v) ethanol | Macrococcus caseolyticus |  |
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| expressed in Escherichia coli ER2566 cells | Macrococcus caseolyticus |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| E50A | the mutant shows reduced activity and FAD binding compared to the wild type enzyme | Macrococcus caseolyticus |
| E56A | inactive | Macrococcus caseolyticus |
| G29A | inactive | Macrococcus caseolyticus |
| G31A | inactive | Macrococcus caseolyticus |
| S34A | the mutant shows reduced activity and FAD binding compared to the wild type enzyme | Macrococcus caseolyticus |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| Ba2+ | - |
Macrococcus caseolyticus | |
| Ca2+ | - |
Macrococcus caseolyticus | |
| Co2+ | - |
Macrococcus caseolyticus | |
| Cu2+ | - |
Macrococcus caseolyticus | |
| Fe2+ | - |
Macrococcus caseolyticus | |
| Mg2+ | - |
Macrococcus caseolyticus | |
| Mn2+ | - |
Macrococcus caseolyticus | |
| additional information | not inhibited by EDTA | Macrococcus caseolyticus | |
| Ni2+ | - |
Macrococcus caseolyticus | |
| Zn2+ | - |
Macrococcus caseolyticus | |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.058 | - |
linoleic acid | at pH 6.5 and 25°C | Macrococcus caseolyticus | |
| 0.059 | - |
alpha-linolenic acid | at pH 6.5 and 25°C | Macrococcus caseolyticus | |
| 0.3 | - |
myristoleic acid | at pH 6.5 and 25°C | Macrococcus caseolyticus | |
| 0.34 | - |
oleic acid | at pH 6.5 and 25°C | Macrococcus caseolyticus | |
| 0.57 | - |
palmitoleic acid | at pH 6.5 and 25°C | Macrococcus caseolyticus | |
| 1.12 | - |
gamma-linolenic acid | at pH 6.5 and 25°C | Macrococcus caseolyticus | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 68000 | - |
2 * 68000, SDS-PAGE | Macrococcus caseolyticus |
| 68100 | - |
2 * 68100, calculated from amino acid sequence | Macrococcus caseolyticus |
| 136000 | - |
gel filtration | Macrococcus caseolyticus |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| oleic acid + H2O | Macrococcus caseolyticus | - |
10-hydroxyoctadecanoic acid | - |
ir | |
| oleic acid + H2O | Macrococcus caseolyticus JCSC5402 | - |
10-hydroxyoctadecanoic acid | - |
ir | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Macrococcus caseolyticus | B9E972 | - |
- |
| Macrococcus caseolyticus JCSC5402 | B9E972 | - |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| His-Trap column chromatography | Macrococcus caseolyticus |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 3.3 | - |
at pH 6.5 and 25°C | Macrococcus caseolyticus |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| alpha-linolenic acid + H2O | - |
Macrococcus caseolyticus | 10-hydroxy-12(Z),15(Z)-octadecadienoic acid + 10,13-dihydroxy-15(Z)-octadecenoic acid | - |
? | |
| alpha-linolenic acid + H2O | - |
Macrococcus caseolyticus JCSC5402 | 10-hydroxy-12(Z),15(Z)-octadecadienoic acid + 10,13-dihydroxy-15(Z)-octadecenoic acid | - |
? | |
| gamma-linolenic acid | - |
Macrococcus caseolyticus | 10-hydroxy-6(Z),12(Z)-octadecadienoic acid + 10,13-dihydroxy-6(Z)-octadecenoic acid | - |
? | |
| gamma-linolenic acid | - |
Macrococcus caseolyticus JCSC5402 | 10-hydroxy-6(Z),12(Z)-octadecadienoic acid + 10,13-dihydroxy-6(Z)-octadecenoic acid | - |
? | |
| linoleic acid | - |
Macrococcus caseolyticus | 10-hydroxy-12(Z)-octadecenoic acid + 10,13-dihydroxyoctadecanoic acid | - |
? | |
| linoleic acid | - |
Macrococcus caseolyticus JCSC5402 | 10-hydroxy-12(Z)-octadecenoic acid + 10,13-dihydroxyoctadecanoic acid | - |
? | |
| additional information | no activity with 10-hydroxyoctadecanoic acid | Macrococcus caseolyticus | ? | - |
? | |
| additional information | no activity with 10-hydroxyoctadecanoic acid | Macrococcus caseolyticus JCSC5402 | ? | - |
? | |
| myristoleic acid | - |
Macrococcus caseolyticus | 10-hydroxytetradecanoic acid + H2O | - |
? | |
| oleic acid + H2O | - |
Macrococcus caseolyticus | 10-hydroxyoctadecanoic acid | - |
ir | |
| oleic acid + H2O | - |
Macrococcus caseolyticus JCSC5402 | 10-hydroxyoctadecanoic acid | - |
ir | |
| palmitoleic acid | - |
Macrococcus caseolyticus | 10-hydroxyhexadecanoic acid + H2O | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| homodimer | 2 * 68000, SDS-PAGE | Macrococcus caseolyticus |
| homodimer | 2 * 68100, calculated from amino acid sequence | Macrococcus caseolyticus |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 25 | - |
- |
Macrococcus caseolyticus |
Temperature Range [°C]
| Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 15 | 35 | 40% activity at 15°C, 75% activity at 20°C, 100% activity at 25°C, 78% activity at 30°C, and 65% activity at 35°C | Macrococcus caseolyticus |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 25 | 35 | after 120 min, the enzyme activity is nearly unchanges at 25°C, but is reduced to only 25% at 35 °C | Macrococcus caseolyticus |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.03 | - |
myristoleic acid | at pH 6.5 and 25°C | Macrococcus caseolyticus | |
| 0.13 | - |
linoleic acid | at pH 6.5 and 25°C | Macrococcus caseolyticus | |
| 0.13 | - |
alpha-linolenic acid | at pH 6.5 and 25°C | Macrococcus caseolyticus | |
| 2.67 | - |
gamma-linolenic acid | at pH 6.5 and 25°C | Macrococcus caseolyticus | |
| 3.83 | - |
palmitoleic acid | at pH 6.5 and 25°C | Macrococcus caseolyticus | |
| 7.83 | - |
oleic acid | at pH 6.5 and 25°C | Macrococcus caseolyticus | |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 6.5 | - |
- |
Macrococcus caseolyticus |
pH Range
| pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|
| 5.5 | 7.5 | 48% activity at pH 5.5, 68% activity at pH 6.0, 100% activity at pH 6.5, 40% activity at pH 7.0, and 20% activity at pH 7.5 | Macrococcus caseolyticus |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| FAD | required for activity, maximum activity in the presence of 0.2 M FAD | Macrococcus caseolyticus | |
| additional information | the addition of the cofactor FMN, NAD+, or NADP+ does not affect enzyme activity | Macrococcus caseolyticus |
kcat/KM [mM/s]
| kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.13 | - |
myristoleic acid | at pH 6.5 and 25°C | Macrococcus caseolyticus | |
| 0.33 | - |
oleic acid | at pH 6.5 and 25°C | Macrococcus caseolyticus | |
| 0.33 | - |
linoleic acid | at pH 6.5 and 25°C | Macrococcus caseolyticus | |
| 0.42 | - |
alpha-linolenic acid | at pH 6.5 and 25°C | Macrococcus caseolyticus | |
| 4.5 | - |
gamma-linolenic acid | at pH 6.5 and 25°C | Macrococcus caseolyticus | |
| 22.67 | - |
palmitoleic acid | at pH 6.5 and 25°C | Macrococcus caseolyticus | |
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