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Literature summary extracted from

  • Joo, Y.C.; Jeong, K.W.; Yeom, S.J.; Kim, Y.S.; Kim, Y.; Oh, D.K.
    Biochemical characterization and FAD-binding analysis of oleate hydratase from Macrococcus caseolyticus (2012), Biochimie, 94, 907-915.
    View publication on PubMed

Activating Compound

EC Number Activating Compound Comment Organism Structure
4.2.1.53 ethanol maximum activity in the presence of 2% (v/v) ethanol Macrococcus caseolyticus

Cloned(Commentary)

EC Number Cloned (Comment) Organism
4.2.1.53 expressed in Escherichia coli ER2566 cells Macrococcus caseolyticus

Protein Variants

EC Number Protein Variants Comment Organism
4.2.1.53 E50A the mutant shows reduced activity and FAD binding compared to the wild type enzyme Macrococcus caseolyticus
4.2.1.53 E56A inactive Macrococcus caseolyticus
4.2.1.53 G29A inactive Macrococcus caseolyticus
4.2.1.53 G31A inactive Macrococcus caseolyticus
4.2.1.53 S34A the mutant shows reduced activity and FAD binding compared to the wild type enzyme Macrococcus caseolyticus

Inhibitors

EC Number Inhibitors Comment Organism Structure
4.2.1.53 Ba2+
-
 Macrococcus caseolyticus
4.2.1.53 Ca2+
-
 Macrococcus caseolyticus
4.2.1.53 Co2+
-
 Macrococcus caseolyticus
4.2.1.53 Cu2+
-
 Macrococcus caseolyticus
4.2.1.53 Fe2+
-
 Macrococcus caseolyticus
4.2.1.53 Mg2+
-
 Macrococcus caseolyticus
4.2.1.53 Mn2+
-
 Macrococcus caseolyticus
4.2.1.53 additional information not inhibited by EDTA Macrococcus caseolyticus
4.2.1.53 Ni2+
-
 Macrococcus caseolyticus
4.2.1.53 Zn2+
-
 Macrococcus caseolyticus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
4.2.1.53 0.058
-
 linoleic acid at pH 6.5 and 25°C Macrococcus caseolyticus
4.2.1.53 0.059
-
 alpha-linolenic acid at pH 6.5 and 25°C Macrococcus caseolyticus
4.2.1.53 0.3
-
 myristoleic acid at pH 6.5 and 25°C Macrococcus caseolyticus
4.2.1.53 0.34
-
 oleic acid at pH 6.5 and 25°C Macrococcus caseolyticus
4.2.1.53 0.57
-
 palmitoleic acid at pH 6.5 and 25°C Macrococcus caseolyticus
4.2.1.53 1.12
-
 gamma-linolenic acid at pH 6.5 and 25°C Macrococcus caseolyticus

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
4.2.1.53 68000
-
 2 * 68000, SDS-PAGE Macrococcus caseolyticus
4.2.1.53 68100
-
 2 * 68100, calculated from amino acid sequence Macrococcus caseolyticus
4.2.1.53 136000
-
 gel filtration Macrococcus caseolyticus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
4.2.1.53 oleic acid + H2O Macrococcus caseolyticus
-
 10-hydroxyoctadecanoic acid
-
 ir
4.2.1.53 oleic acid + H2O Macrococcus caseolyticus JCSC5402
-
 10-hydroxyoctadecanoic acid
-
 ir

Organism

EC Number Organism UniProt Comment Textmining
4.2.1.53 Macrococcus caseolyticus B9E972
-
-
4.2.1.53 Macrococcus caseolyticus JCSC5402 B9E972
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
4.2.1.53 His-Trap column chromatography Macrococcus caseolyticus

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
4.2.1.53 3.3
-
 at pH 6.5 and 25°C Macrococcus caseolyticus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.2.1.53 alpha-linolenic acid + H2O
-
 Macrococcus caseolyticus 10-hydroxy-12(Z),15(Z)-octadecadienoic acid + 10,13-dihydroxy-15(Z)-octadecenoic acid
-
 ?
4.2.1.53 alpha-linolenic acid + H2O
-
 Macrococcus caseolyticus JCSC5402 10-hydroxy-12(Z),15(Z)-octadecadienoic acid + 10,13-dihydroxy-15(Z)-octadecenoic acid
-
 ?
4.2.1.53 gamma-linolenic acid
-
 Macrococcus caseolyticus 10-hydroxy-6(Z),12(Z)-octadecadienoic acid + 10,13-dihydroxy-6(Z)-octadecenoic acid
-
 ?
4.2.1.53 gamma-linolenic acid
-
 Macrococcus caseolyticus JCSC5402 10-hydroxy-6(Z),12(Z)-octadecadienoic acid + 10,13-dihydroxy-6(Z)-octadecenoic acid
-
 ?
4.2.1.53 linoleic acid
-
 Macrococcus caseolyticus 10-hydroxy-12(Z)-octadecenoic acid + 10,13-dihydroxyoctadecanoic acid
-
 ?
4.2.1.53 linoleic acid
-
 Macrococcus caseolyticus JCSC5402 10-hydroxy-12(Z)-octadecenoic acid + 10,13-dihydroxyoctadecanoic acid
-
 ?
4.2.1.53 additional information no activity with 10-hydroxyoctadecanoic acid Macrococcus caseolyticus ?
-
 ?
4.2.1.53 additional information no activity with 10-hydroxyoctadecanoic acid Macrococcus caseolyticus JCSC5402 ?
-
 ?
4.2.1.53 myristoleic acid
-
 Macrococcus caseolyticus 10-hydroxytetradecanoic acid + H2O
-
 ?
4.2.1.53 oleic acid + H2O
-
 Macrococcus caseolyticus 10-hydroxyoctadecanoic acid
-
 ir
4.2.1.53 oleic acid + H2O
-
 Macrococcus caseolyticus JCSC5402 10-hydroxyoctadecanoic acid
-
 ir
4.2.1.53 palmitoleic acid
-
 Macrococcus caseolyticus 10-hydroxyhexadecanoic acid + H2O
-
 ?

Subunits

EC Number Subunits Comment Organism
4.2.1.53 homodimer 2 * 68000, SDS-PAGE Macrococcus caseolyticus
4.2.1.53 homodimer 2 * 68100, calculated from amino acid sequence Macrococcus caseolyticus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
4.2.1.53 25
-
-
 Macrococcus caseolyticus

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
4.2.1.53 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]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
4.2.1.53 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]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
4.2.1.53 0.03
-
 myristoleic acid at pH 6.5 and 25°C Macrococcus caseolyticus
4.2.1.53 0.13
-
 linoleic acid at pH 6.5 and 25°C Macrococcus caseolyticus
4.2.1.53 0.13
-
 alpha-linolenic acid at pH 6.5 and 25°C Macrococcus caseolyticus
4.2.1.53 2.67
-
 gamma-linolenic acid at pH 6.5 and 25°C Macrococcus caseolyticus
4.2.1.53 3.83
-
 palmitoleic acid at pH 6.5 and 25°C Macrococcus caseolyticus
4.2.1.53 7.83
-
 oleic acid at pH 6.5 and 25°C Macrococcus caseolyticus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
4.2.1.53 6.5
-
-
 Macrococcus caseolyticus

pH Range

EC Number pH Minimum pH Maximum Comment Organism
4.2.1.53 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

EC Number Cofactor Comment Organism Structure
4.2.1.53 FAD required for activity, maximum activity in the presence of 0.2 M FAD Macrococcus caseolyticus
4.2.1.53 additional information the addition of the cofactor FMN, NAD+, or NADP+ does not affect enzyme activity Macrococcus caseolyticus

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
4.2.1.53 0.13
-
 myristoleic acid at pH 6.5 and 25°C Macrococcus caseolyticus
4.2.1.53 0.33
-
 oleic acid at pH 6.5 and 25°C Macrococcus caseolyticus
4.2.1.53 0.33
-
 linoleic acid at pH 6.5 and 25°C Macrococcus caseolyticus
4.2.1.53 0.42
-
 alpha-linolenic acid at pH 6.5 and 25°C Macrococcus caseolyticus
4.2.1.53 4.5
-
 gamma-linolenic acid at pH 6.5 and 25°C Macrococcus caseolyticus
4.2.1.53 22.67
-
 palmitoleic acid at pH 6.5 and 25°C Macrococcus caseolyticus