BRENDA - Enzyme Database

Biochemical and structural characterization of CYP124: a methyl-branched lipid omega-hydroxylase from Mycobacterium tuberculosis

Johnston, J.B.; Kells, P.M.; Podust, L.M.; Ortiz de Montellano, P.R.; Proc. Natl. Acad. Sci. USA 106, 20687-20692 (2009)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
1.14.15.14
expressed in Escherichia coli DH5alpha cells
Mycobacterium tuberculosis
Crystallization (Commentary)
EC Number
Crystallization
Organism
1.14.15.14
hanging drop vapor diffusion method, free enzyme using 1% (w/v) PEGMME 2000, 0.1 M Na cacodylate (pH 6.8), and 0.9 M succinic acid, and enzyme in complex with phytanic acid using 21% (w/v) PEG 4000, 0.1 M Bis-Tris (pH 5.5), and 0.2 M calcium acetate
Mycobacterium tuberculosis
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.14.15.14
0.009
-
15-methylpalmitic acid
at pH 7.4 and 25C
Mycobacterium tuberculosis
1.14.15.14
0.032
-
geranylgeraniol
at pH 7.4 and 25C
Mycobacterium tuberculosis
1.14.15.14
0.036
-
farnesol
at pH 7.4 and 25C
Mycobacterium tuberculosis
1.14.15.14
0.054
-
phytanic acid
at pH 7.4 and 25C
Mycobacterium tuberculosis
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
1.14.15.14
48700
-
x * 48700, calculated from amino acid sequence
Mycobacterium tuberculosis
1.14.15.14
50000
-
x * 50000, SDS-PAGE
Mycobacterium tuberculosis
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.14.15.14
a methyl-branched lipid + O2 + reduced ferredoxin [iron-sulfur] cluster + H+
Mycobacterium tuberculosis
-
an omega-hydroxy-methyl-branched lipid + H2O + oxidized ferredoxin [iron-sulfur] cluster
-
-
?
1.14.15.14
a methyl-branched lipid + O2 + reduced ferredoxin [iron-sulfur] cluster + H+
Mycobacterium tuberculosis H37Rv
-
an omega-hydroxy-methyl-branched lipid + H2O + oxidized ferredoxin [iron-sulfur] cluster
-
-
?
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
1.14.15.14
Mycobacterium tuberculosis
P9WPP3
-
-
1.14.15.14
Mycobacterium tuberculosis H37Rv
P9WPP3
-
-
Purification (Commentary)
EC Number
Commentary
Organism
1.14.15.14
Ni2+-NTA column chromatography
Mycobacterium tuberculosis
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.14.15.14
15-methylpalmitic acid + O2 + reduced ferredoxin [iron-sulfur] cluster + H+
-
735155
Mycobacterium tuberculosis
?
-
-
-
?
1.14.15.14
15-methylpalmitic acid + O2 + reduced ferredoxin [iron-sulfur] cluster + H+
-
735155
Mycobacterium tuberculosis H37Rv
?
-
-
-
?
1.14.15.14
a methyl-branched lipid + O2 + reduced ferredoxin [iron-sulfur] cluster + H+
-
735155
Mycobacterium tuberculosis
an omega-hydroxy-methyl-branched lipid + H2O + oxidized ferredoxin [iron-sulfur] cluster
-
-
-
?
1.14.15.14
a methyl-branched lipid + O2 + reduced ferredoxin [iron-sulfur] cluster + H+
-
735155
Mycobacterium tuberculosis H37Rv
an omega-hydroxy-methyl-branched lipid + H2O + oxidized ferredoxin [iron-sulfur] cluster
-
-
-
?
1.14.15.14
farnesol + O2 + reduced ferredoxin [iron-sulfur] cluster + H+
best substrate
735155
Mycobacterium tuberculosis
?
-
-
-
?
1.14.15.14
farnesol + O2 + reduced ferredoxin [iron-sulfur] cluster + H+
best substrate
735155
Mycobacterium tuberculosis H37Rv
?
-
-
-
?
1.14.15.14
farnesyl diphosphate + O2 + reduced ferredoxin [iron-sulfur] cluster + H+
-
735155
Mycobacterium tuberculosis
?
-
-
-
?
1.14.15.14
farnesyl diphosphate + O2 + reduced ferredoxin [iron-sulfur] cluster + H+
-
735155
Mycobacterium tuberculosis H37Rv
?
-
-
-
?
1.14.15.14
geranylgeraniol + O2 + reduced ferredoxin [iron-sulfur] cluster + H+
-
735155
Mycobacterium tuberculosis
?
-
-
-
?
1.14.15.14
additional information
the enzyme shows no activity with geraniol, lauric acid, phytane, pristane, phylloquinone (vitamin K1), menaquinone (vitamin K2), or bifonazole
735155
Mycobacterium tuberculosis
?
-
-
-
-
1.14.15.14
additional information
the enzyme shows no activity with geraniol, lauric acid, phytane, pristane, phylloquinone (vitamin K1), menaquinone (vitamin K2), or bifonazole
735155
Mycobacterium tuberculosis H37Rv
?
-
-
-
-
1.14.15.14
palmitic acid + O2 + reduced ferredoxin [iron-sulfur] cluster + H+
the enzyme oxidizes the omega-1,omega-2, and omega-position of palmitic acid
735155
Mycobacterium tuberculosis
?
-
-
-
?
1.14.15.14
phytanic acid + O2 + reduced ferredoxin [iron-sulfur] cluster + H+
-
735155
Mycobacterium tuberculosis
?
-
-
-
?
Subunits
EC Number
Subunits
Commentary
Organism
1.14.15.14
?
x * 48700, calculated from amino acid sequence; x * 50000, SDS-PAGE
Mycobacterium tuberculosis
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
1.14.15.14
expressed in Escherichia coli DH5alpha cells
Mycobacterium tuberculosis
Crystallization (Commentary) (protein specific)
EC Number
Crystallization
Organism
1.14.15.14
hanging drop vapor diffusion method, free enzyme using 1% (w/v) PEGMME 2000, 0.1 M Na cacodylate (pH 6.8), and 0.9 M succinic acid, and enzyme in complex with phytanic acid using 21% (w/v) PEG 4000, 0.1 M Bis-Tris (pH 5.5), and 0.2 M calcium acetate
Mycobacterium tuberculosis
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.14.15.14
0.009
-
15-methylpalmitic acid
at pH 7.4 and 25C
Mycobacterium tuberculosis
1.14.15.14
0.032
-
geranylgeraniol
at pH 7.4 and 25C
Mycobacterium tuberculosis
1.14.15.14
0.036
-
farnesol
at pH 7.4 and 25C
Mycobacterium tuberculosis
1.14.15.14
0.054
-
phytanic acid
at pH 7.4 and 25C
Mycobacterium tuberculosis
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
1.14.15.14
48700
-
x * 48700, calculated from amino acid sequence
Mycobacterium tuberculosis
1.14.15.14
50000
-
x * 50000, SDS-PAGE
Mycobacterium tuberculosis
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.14.15.14
a methyl-branched lipid + O2 + reduced ferredoxin [iron-sulfur] cluster + H+
Mycobacterium tuberculosis
-
an omega-hydroxy-methyl-branched lipid + H2O + oxidized ferredoxin [iron-sulfur] cluster
-
-
?
1.14.15.14
a methyl-branched lipid + O2 + reduced ferredoxin [iron-sulfur] cluster + H+
Mycobacterium tuberculosis H37Rv
-
an omega-hydroxy-methyl-branched lipid + H2O + oxidized ferredoxin [iron-sulfur] cluster
-
-
?
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
1.14.15.14
Ni2+-NTA column chromatography
Mycobacterium tuberculosis
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.14.15.14
15-methylpalmitic acid + O2 + reduced ferredoxin [iron-sulfur] cluster + H+
-
735155
Mycobacterium tuberculosis
?
-
-
-
?
1.14.15.14
15-methylpalmitic acid + O2 + reduced ferredoxin [iron-sulfur] cluster + H+
-
735155
Mycobacterium tuberculosis H37Rv
?
-
-
-
?
1.14.15.14
a methyl-branched lipid + O2 + reduced ferredoxin [iron-sulfur] cluster + H+
-
735155
Mycobacterium tuberculosis
an omega-hydroxy-methyl-branched lipid + H2O + oxidized ferredoxin [iron-sulfur] cluster
-
-
-
?
1.14.15.14
a methyl-branched lipid + O2 + reduced ferredoxin [iron-sulfur] cluster + H+
-
735155
Mycobacterium tuberculosis H37Rv
an omega-hydroxy-methyl-branched lipid + H2O + oxidized ferredoxin [iron-sulfur] cluster
-
-
-
?
1.14.15.14
farnesol + O2 + reduced ferredoxin [iron-sulfur] cluster + H+
best substrate
735155
Mycobacterium tuberculosis
?
-
-
-
?
1.14.15.14
farnesol + O2 + reduced ferredoxin [iron-sulfur] cluster + H+
best substrate
735155
Mycobacterium tuberculosis H37Rv
?
-
-
-
?
1.14.15.14
farnesyl diphosphate + O2 + reduced ferredoxin [iron-sulfur] cluster + H+
-
735155
Mycobacterium tuberculosis
?
-
-
-
?
1.14.15.14
farnesyl diphosphate + O2 + reduced ferredoxin [iron-sulfur] cluster + H+
-
735155
Mycobacterium tuberculosis H37Rv
?
-
-
-
?
1.14.15.14
geranylgeraniol + O2 + reduced ferredoxin [iron-sulfur] cluster + H+
-
735155
Mycobacterium tuberculosis
?
-
-
-
?
1.14.15.14
additional information
the enzyme shows no activity with geraniol, lauric acid, phytane, pristane, phylloquinone (vitamin K1), menaquinone (vitamin K2), or bifonazole
735155
Mycobacterium tuberculosis
?
-
-
-
-
1.14.15.14
additional information
the enzyme shows no activity with geraniol, lauric acid, phytane, pristane, phylloquinone (vitamin K1), menaquinone (vitamin K2), or bifonazole
735155
Mycobacterium tuberculosis H37Rv
?
-
-
-
-
1.14.15.14
palmitic acid + O2 + reduced ferredoxin [iron-sulfur] cluster + H+
the enzyme oxidizes the omega-1,omega-2, and omega-position of palmitic acid
735155
Mycobacterium tuberculosis
?
-
-
-
?
1.14.15.14
phytanic acid + O2 + reduced ferredoxin [iron-sulfur] cluster + H+
-
735155
Mycobacterium tuberculosis
?
-
-
-
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
1.14.15.14
?
x * 48700, calculated from amino acid sequence; x * 50000, SDS-PAGE
Mycobacterium tuberculosis