BRENDA - Enzyme Database show

Isolation and characterization of a lipoxygenase from Pseudomonas 42A2 responsible for the biotransformation of oleic acid into (S)-(E)-10-hydroxy-8-octadecenoic acid

Busquets, M.; Deroncelé, V.; Vidal-Mas, J.; Rodríguez, E.; Guerrero, A.; Manresa, A.; Antonie van Leeuwenhoek 85, 129-139 (2004)

Data extracted from this reference:

Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
1.13.11.77
Ba2+
complete inhibition at 1 mM
Pseudomonas sp.
1.13.11.77
EDTA
99% inhibition at 5 mM
Pseudomonas sp.
1.13.11.77
Fe3+
complete inhibition at 1 mM
Pseudomonas sp.
1.13.11.77
Mg2+
concentrations of Mg2+ higher than 1.25 mM inhibit activity strongly (90% of activity lost at 1.75 mM)
Pseudomonas sp.
1.13.11.77
additional information
not inhibited by 5 mM KCN
Pseudomonas sp.
1.13.11.77
oleate
at concentrations of oleate higher than 2.1 mM, the enzyme activity is inhibited by excess of substrate
Pseudomonas sp.
1.13.11.77
Zn2+
complete inhibition at 1 mM
Pseudomonas sp.
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.13.11.77
0.66
-
linoleate
in 0.2 M sodium borate buffer, pH 9.0, at 30°C
Pseudomonas sp.
1.13.11.77
0.73
-
alpha-linolenate
in 0.2 M sodium borate buffer, pH 9.0, at 30°C
Pseudomonas sp.
1.13.11.77
0.74
-
oleate
in 0.2 M sodium borate buffer, pH 9.0, at 30°C
Pseudomonas sp.
Localization
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
1.13.11.77
periplasm
-
Pseudomonas sp.
-
-
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
1.13.11.77
Fe2+
the enzyme contains 0.55 mol of Fe2+ per mol of protein
Pseudomonas sp.
1.13.11.77
Mg2+
the presence of 1 mM Mg2+ increases enzyme activity 1.5fold
Pseudomonas sp.
1.13.11.77
Mn2+
the enzyme contains 25 mmol Mn2+ per mol of protein
Pseudomonas sp.
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
1.13.11.77
45000
-
1 * 45000, SDS-PAGE
Pseudomonas sp.
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.13.11.77
linoleate + O2
Pseudomonas sp.
100% activity
(8E,10S,12Z)-10-hydroperoxyoctadeca-8,12-dienoate
-
-
?
1.13.11.77
linoleate + O2
Pseudomonas sp. 42A2
100% activity
(8E,10S,12Z)-10-hydroperoxyoctadeca-8,12-dienoate
-
-
?
1.13.11.77
oleate + O2
Pseudomonas sp.
46% activity compared to linoleate
(8E,10S)-10-hydroperoxyoctadeca-8-enoate
-
-
?
1.13.11.77
oleate + O2
Pseudomonas sp. 42A2
46% activity compared to linoleate
(8E,10S)-10-hydroperoxyoctadeca-8-enoate
-
-
?
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
1.13.11.77
Pseudomonas sp.
-
-
-
1.13.11.77
Pseudomonas sp. 42A2
-
-
-
Purification (Commentary)
EC Number
Commentary
Organism
1.13.11.77
Q Sepharose column chromatography and Macro-Prep SE 100/40 gel filtration
Pseudomonas sp.
Storage Stability
EC Number
Storage Stability
Organism
1.13.11.77
4°C, 1 week, no loss of activity (thawing and freezing decreases enzyme activity by 30%)
Pseudomonas sp.
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.13.11.77
alpha-linolenate + O2
60.4% activity compared to linoleate
723962
Pseudomonas sp.
(8E,10S,12Z,15Z)-10-hydroperoxyoctadeca-8,12,15-trienoate
-
-
-
?
1.13.11.77
alpha-linolenate + O2
60.4% activity compared to linoleate
723962
Pseudomonas sp. 42A2
(8E,10S,12Z,15Z)-10-hydroperoxyoctadeca-8,12,15-trienoate
-
-
-
?
1.13.11.77
arachidonate + O2
0.9% activity compared to linoleate
723962
Pseudomonas sp.
?
-
-
-
?
1.13.11.77
arachidonate + O2
0.9% activity compared to linoleate
723962
Pseudomonas sp. 42A2
?
-
-
-
?
1.13.11.77
cis-12-octadecenoate + O2
8.9% activity compared to linoleate
723962
Pseudomonas sp.
?
-
-
-
?
1.13.11.77
linoleate + O2
100% activity
723962
Pseudomonas sp.
(8E,10S,12Z)-10-hydroperoxyoctadeca-8,12-dienoate
-
-
-
?
1.13.11.77
linoleate + O2
100% activity
723962
Pseudomonas sp. 42A2
(8E,10S,12Z)-10-hydroperoxyoctadeca-8,12-dienoate
-
-
-
?
1.13.11.77
additional information
the activity of the enzyme is highest in unsaturated fatty acids containing double bonds in position 9 (oleic, linoleic and linolenic acids), linoleic acid being preferred over linolenic and oleic acids
723962
Pseudomonas sp.
?
-
-
-
-
1.13.11.77
additional information
the activity of the enzyme is highest in unsaturated fatty acids containing double bonds in position 9 (oleic, linoleic and linolenic acids), linoleic acid being preferred over linolenic and oleic acids
723962
Pseudomonas sp. 42A2
?
-
-
-
-
1.13.11.77
oleate + O2
46% activity compared to linoleate
723962
Pseudomonas sp.
(8E,10S)-10-hydroperoxyoctadeca-8-enoate
-
-
-
?
1.13.11.77
oleate + O2
46% activity compared to linoleate
723962
Pseudomonas sp. 42A2
(8E,10S)-10-hydroperoxyoctadeca-8-enoate
-
-
-
?
1.13.11.77
petroselinoate + O2
26.4% activity compared to linoleate
723962
Pseudomonas sp.
?
-
-
-
?
1.13.11.77
triolein + O2
0.4% activity compared to linoleate
723962
Pseudomonas sp.
?
-
-
-
?
Subunits
EC Number
Subunits
Commentary
Organism
1.13.11.77
monomer
1 * 45000, SDS-PAGE
Pseudomonas sp.
Temperature Optimum [°C]
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
1.13.11.77
25
30
-
Pseudomonas sp.
Temperature Stability [°C]
EC Number
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
1.13.11.77
45
70
44% of activity still remains at 55°C. The enzyme is stable up to 45 °C, but sharply decreases after 50°C. Activity is completely lost at 70°C
Pseudomonas sp.
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.13.11.77
8.5
9
-
Pseudomonas sp.
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
1.13.11.77
Ba2+
complete inhibition at 1 mM
Pseudomonas sp.
1.13.11.77
EDTA
99% inhibition at 5 mM
Pseudomonas sp.
1.13.11.77
Fe3+
complete inhibition at 1 mM
Pseudomonas sp.
1.13.11.77
Mg2+
concentrations of Mg2+ higher than 1.25 mM inhibit activity strongly (90% of activity lost at 1.75 mM)
Pseudomonas sp.
1.13.11.77
additional information
not inhibited by 5 mM KCN
Pseudomonas sp.
1.13.11.77
oleate
at concentrations of oleate higher than 2.1 mM, the enzyme activity is inhibited by excess of substrate
Pseudomonas sp.
1.13.11.77
Zn2+
complete inhibition at 1 mM
Pseudomonas sp.
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.13.11.77
0.66
-
linoleate
in 0.2 M sodium borate buffer, pH 9.0, at 30°C
Pseudomonas sp.
1.13.11.77
0.73
-
alpha-linolenate
in 0.2 M sodium borate buffer, pH 9.0, at 30°C
Pseudomonas sp.
1.13.11.77
0.74
-
oleate
in 0.2 M sodium borate buffer, pH 9.0, at 30°C
Pseudomonas sp.
Localization (protein specific)
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
1.13.11.77
periplasm
-
Pseudomonas sp.
-
-
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
1.13.11.77
Fe2+
the enzyme contains 0.55 mol of Fe2+ per mol of protein
Pseudomonas sp.
1.13.11.77
Mg2+
the presence of 1 mM Mg2+ increases enzyme activity 1.5fold
Pseudomonas sp.
1.13.11.77
Mn2+
the enzyme contains 25 mmol Mn2+ per mol of protein
Pseudomonas sp.
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
1.13.11.77
45000
-
1 * 45000, SDS-PAGE
Pseudomonas sp.
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.13.11.77
linoleate + O2
Pseudomonas sp.
100% activity
(8E,10S,12Z)-10-hydroperoxyoctadeca-8,12-dienoate
-
-
?
1.13.11.77
linoleate + O2
Pseudomonas sp. 42A2
100% activity
(8E,10S,12Z)-10-hydroperoxyoctadeca-8,12-dienoate
-
-
?
1.13.11.77
oleate + O2
Pseudomonas sp.
46% activity compared to linoleate
(8E,10S)-10-hydroperoxyoctadeca-8-enoate
-
-
?
1.13.11.77
oleate + O2
Pseudomonas sp. 42A2
46% activity compared to linoleate
(8E,10S)-10-hydroperoxyoctadeca-8-enoate
-
-
?
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
1.13.11.77
Q Sepharose column chromatography and Macro-Prep SE 100/40 gel filtration
Pseudomonas sp.
Storage Stability (protein specific)
EC Number
Storage Stability
Organism
1.13.11.77
4°C, 1 week, no loss of activity (thawing and freezing decreases enzyme activity by 30%)
Pseudomonas sp.
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.13.11.77
alpha-linolenate + O2
60.4% activity compared to linoleate
723962
Pseudomonas sp.
(8E,10S,12Z,15Z)-10-hydroperoxyoctadeca-8,12,15-trienoate
-
-
-
?
1.13.11.77
alpha-linolenate + O2
60.4% activity compared to linoleate
723962
Pseudomonas sp. 42A2
(8E,10S,12Z,15Z)-10-hydroperoxyoctadeca-8,12,15-trienoate
-
-
-
?
1.13.11.77
arachidonate + O2
0.9% activity compared to linoleate
723962
Pseudomonas sp.
?
-
-
-
?
1.13.11.77
arachidonate + O2
0.9% activity compared to linoleate
723962
Pseudomonas sp. 42A2
?
-
-
-
?
1.13.11.77
cis-12-octadecenoate + O2
8.9% activity compared to linoleate
723962
Pseudomonas sp.
?
-
-
-
?
1.13.11.77
linoleate + O2
100% activity
723962
Pseudomonas sp.
(8E,10S,12Z)-10-hydroperoxyoctadeca-8,12-dienoate
-
-
-
?
1.13.11.77
linoleate + O2
100% activity
723962
Pseudomonas sp. 42A2
(8E,10S,12Z)-10-hydroperoxyoctadeca-8,12-dienoate
-
-
-
?
1.13.11.77
additional information
the activity of the enzyme is highest in unsaturated fatty acids containing double bonds in position 9 (oleic, linoleic and linolenic acids), linoleic acid being preferred over linolenic and oleic acids
723962
Pseudomonas sp.
?
-
-
-
-
1.13.11.77
additional information
the activity of the enzyme is highest in unsaturated fatty acids containing double bonds in position 9 (oleic, linoleic and linolenic acids), linoleic acid being preferred over linolenic and oleic acids
723962
Pseudomonas sp. 42A2
?
-
-
-
-
1.13.11.77
oleate + O2
46% activity compared to linoleate
723962
Pseudomonas sp.
(8E,10S)-10-hydroperoxyoctadeca-8-enoate
-
-
-
?
1.13.11.77
oleate + O2
46% activity compared to linoleate
723962
Pseudomonas sp. 42A2
(8E,10S)-10-hydroperoxyoctadeca-8-enoate
-
-
-
?
1.13.11.77
petroselinoate + O2
26.4% activity compared to linoleate
723962
Pseudomonas sp.
?
-
-
-
?
1.13.11.77
triolein + O2
0.4% activity compared to linoleate
723962
Pseudomonas sp.
?
-
-
-
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
1.13.11.77
monomer
1 * 45000, SDS-PAGE
Pseudomonas sp.
Temperature Optimum [°C] (protein specific)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
1.13.11.77
25
30
-
Pseudomonas sp.
Temperature Stability [°C] (protein specific)
EC Number
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
1.13.11.77
45
70
44% of activity still remains at 55°C. The enzyme is stable up to 45 °C, but sharply decreases after 50°C. Activity is completely lost at 70°C
Pseudomonas sp.
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.13.11.77
8.5
9
-
Pseudomonas sp.