BRENDA - Enzyme Database

Purification and properties of cobalamin-independent methionine synthase from Candida albicans and Saccharomyces cerevisiae

Suliman, H.S.; Sawyer, G.M.; Appling, D.R.; Robertus, J.D.; Arch. Biochem. Biophys. 441, 56-63 (2005)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
2.1.1.14
expressed in Saccharomyces cerevisiae strain DAY4.1
Candida albicans
2.1.1.14
expressed in Saccharomyces cerevisiae strain DAY4.1
Saccharomyces cerevisiae
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
2.1.1.14
0.013
-
L-homocysteine
in 50 mM potassium phosphate (pH 7.2), 50 mM Tris-Cl (pH 7.2) containing 0.1 mM MgSO4, 10 mM dithiothreitol and 2 mM L-homocysteine at 37°C
Saccharomyces cerevisiae
2.1.1.14
0.014
-
L-homocysteine
in 50 mM potassium phosphate (pH 7.2), 50 mM Tris-Cl (pH 7.2) containing 0.1 mM MgSO4, 10 mM dithiothreitol and 2 mM L-homocysteine at 37°C
Candida albicans
2.1.1.14
0.084
-
5-methyltetrahydropteroyl-gamma-glutamyl-gamma-glutamylglutamate
in 50 mM potassium phosphate (pH 7.2), 50 mM Tris-Cl (pH 7.2) containing 0.1 mM MgSO4, 10 mM dithiothreitol and 2 mM L-homocysteine at 37°C
Saccharomyces cerevisiae
2.1.1.14
0.094
-
5-methyltetrahydropteroyl-gamma-glutamyl-gamma-glutamyl-gamma-glutamylglutamate
in 50 mM potassium phosphate (pH 7.2), 50 mM Tris-Cl (pH 7.2) containing 0.1 mM MgSO4, 10 mM dithiothreitol and 2 mM L-homocysteine at 37°C
Saccharomyces cerevisiae
2.1.1.14
0.109
-
5-methyltetrahydropteroyl-gamma-glutamylglutamate
in 50 mM potassium phosphate (pH 7.2), 50 mM Tris-Cl (pH 7.2) containing 0.1 mM MgSO4, 10 mM dithiothreitol and 2 mM L-homocysteine at 37°C
Saccharomyces cerevisiae
2.1.1.14
0.113
-
5-methyltetrahydropteroyl-gamma-glutamylglutamate
in 50 mM potassium phosphate (pH 7.2), 50 mM Tris-Cl (pH 7.2) containing 0.1 mM MgSO4, 10 mM dithiothreitol and 2 mM L-homocysteine at 37°C
Candida albicans
2.1.1.14
0.12
-
5-methyltetrahydropteroyl-gamma-glutamyl-gamma-glutamyl-gamma-glutamylglutamate
in 50 mM potassium phosphate (pH 7.2), 50 mM Tris-Cl (pH 7.2) containing 0.1 mM MgSO4, 10 mM dithiothreitol and 2 mM L-homocysteine at 37°C
Candida albicans
2.1.1.14
0.129
-
5-methyltetrahydropteroyl-gamma-glutamyl-gamma-glutamylglutamate
in 50 mM potassium phosphate (pH 7.2), 50 mM Tris-Cl (pH 7.2) containing 0.1 mM MgSO4, 10 mM dithiothreitol and 2 mM L-homocysteine at 37°C
Candida albicans
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
2.1.1.14
Candida albicans
-
-
-
2.1.1.14
Saccharomyces cerevisiae
-
-
-
Purification (Commentary)
EC Number
Commentary
Organism
2.1.1.14
DEAE Sepharose column chromatography
Candida albicans
2.1.1.14
HiTrap heparin column chromatography
Saccharomyces cerevisiae
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
2.1.1.14
additional information
-
42.11 units/mg specific activity for 5-methyltetrahydropteroyl-gamma-glutamyl-gamma-glutamylglutamate after 11.7fold purification, in 50 mM potassium phosphate (pH 7.2), 50 mM Tris-Cl (pH 7.2) containing 0.1 mM MgSO4, 10 mM dithiothreitol and 2 mM L-homocysteine at 37°C
Candida albicans
2.1.1.14
additional information
-
29.3 units/mg specific activity for 5-methyltetrahydropteroyl-gamma-glutamyl-gamma-glutamylglutamate after 5.6fold purification, in 50 mM potassium phosphate (pH 7.2), 50 mM Tris-Cl (pH 7.2) containing 0.1 mM MgSO4, 10 mM dithiothreitol and 2 mM L-homocysteine at 37°C
Saccharomyces cerevisiae
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.1.1.14
5-methyltetrahydropteroyl-gamma-glutamyl-gamma-glutamylglutamate + L-homocysteine
-
671587
Saccharomyces cerevisiae
tetrahydropteroyl-gamma-glutamyl-gamma-glutamylglutamate + L-methionine
-
-
-
?
2.1.1.14
5-methyltetrahydropteroyl-gamma-glutamyl-gamma-glutamylglutamate + L-homocysteine
-
671587
Saccharomyces cerevisiae
tetrahydropteroyl-gamma-glutamyl-gamma-glutamylglutamate + L-methionine
-
-
-
r
2.1.1.14
5-methyltetrahydropteroyl-gamma-glutamyl-gamma-glutamylglutamate + L-homocysteine
-
671587
Candida albicans
tetrahydropteroyl-gamma-glutamyl-gamma-glutamylglutamate + L-methionine
-
-
-
?
2.1.1.14
5-methyltetrahydropteroyl-gamma-glutamyl-gamma-glutamylglutamate + L-homocysteine
-
671587
Candida albicans
tetrahydropteroyl-gamma-glutamyl-gamma-glutamylglutamate + L-methionine
-
-
-
r
2.1.1.14
5-methyltetrahydropteroyl-gamma-glutamylglutamate + L-homocysteine
-
671587
Saccharomyces cerevisiae
tetrahydropteroyl-gamma-glutamylglutamate + L-methionine
-
-
-
?
2.1.1.14
5-methyltetrahydropteroyl-gamma-glutamylglutamate + L-homocysteine
-
671587
Candida albicans
tetrahydropteroyl-gamma-glutamylglutamate + L-methionine
-
-
-
?
2.1.1.14
5-methyltetrahydropteroyltri-L-glutamate + L-homocysteine
-
671587
Saccharomyces cerevisiae
tetrahydropteroyltri-L-glutamate + L-methionine
-
-
-
?
2.1.1.14
5-methyltetrahydropteroyltri-L-glutamate + L-homocysteine
-
671587
Candida albicans
tetrahydropteroyltri-L-glutamate + L-methionine
-
-
-
?
2.1.1.14
additional information
does not use 5-methyltetrahydropteroylglutamate as a substrate
671587
Saccharomyces cerevisiae
?
-
-
-
-
2.1.1.14
additional information
does not use 5-methyltetrahydropteroylglutamate as a substrate
671587
Candida albicans
?
-
-
-
-
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
2.1.1.14
0.283
-
5-methyltetrahydropteroyl-gamma-glutamylglutamate
in 50 mM potassium phosphate (pH 7.2), 50 mM Tris-Cl (pH 7.2) containing 0.1 mM MgSO4, 10 mM dithiothreitol and 2 mM L-homocysteine at 37°C
Saccharomyces cerevisiae
2.1.1.14
0.33
-
L-homocysteine
in 50 mM potassium phosphate (pH 7.2), 50 mM Tris-Cl (pH 7.2) containing 0.1 mM MgSO4, 10 mM dithiothreitol and 2 mM L-homocysteine at 37°C
Saccharomyces cerevisiae
2.1.1.14
0.35
-
5-methyltetrahydropteroyl-gamma-glutamylglutamate
in 50 mM potassium phosphate (pH 7.2), 50 mM Tris-Cl (pH 7.2) containing 0.1 mM MgSO4, 10 mM dithiothreitol and 2 mM L-homocysteine at 37°C
Candida albicans
2.1.1.14
0.4
-
5-methyltetrahydropteroyl-gamma-glutamyl-gamma-glutamylglutamate
in 50 mM potassium phosphate (pH 7.2), 50 mM Tris-Cl (pH 7.2) containing 0.1 mM MgSO4, 10 mM dithiothreitol and 2 mM L-homocysteine at 37°C
Candida albicans
2.1.1.14
0.417
-
5-methyltetrahydropteroyl-gamma-glutamyl-gamma-glutamyl-gamma-glutamylglutamate
in 50 mM potassium phosphate (pH 7.2), 50 mM Tris-Cl (pH 7.2) containing 0.1 mM MgSO4, 10 mM dithiothreitol and 2 mM L-homocysteine at 37°C
Candida albicans
2.1.1.14
0.417
-
L-homocysteine
in 50 mM potassium phosphate (pH 7.2), 50 mM Tris-Cl (pH 7.2) containing 0.1 mM MgSO4, 10 mM dithiothreitol and 2 mM L-homocysteine at 37°C
Candida albicans
2.1.1.14
0.533
-
5-methyltetrahydropteroyl-gamma-glutamyl-gamma-glutamylglutamate
in 50 mM potassium phosphate (pH 7.2), 50 mM Tris-Cl (pH 7.2) containing 0.1 mM MgSO4, 10 mM dithiothreitol and 2 mM L-homocysteine at 37°C
Saccharomyces cerevisiae
2.1.1.14
0.6
-
5-methyltetrahydropteroyl-gamma-glutamyl-gamma-glutamyl-gamma-glutamylglutamate
in 50 mM potassium phosphate (pH 7.2), 50 mM Tris-Cl (pH 7.2) containing 0.1 mM MgSO4, 10 mM dithiothreitol and 2 mM L-homocysteine at 37°C
Saccharomyces cerevisiae
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
2.1.1.14
additional information
vitamin-B12 independent enzyme
Candida albicans
2.1.1.14
additional information
vitamin-B12 independent enzyme
Saccharomyces cerevisiae
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
2.1.1.14
expressed in Saccharomyces cerevisiae strain DAY4.1
Candida albicans
2.1.1.14
expressed in Saccharomyces cerevisiae strain DAY4.1
Saccharomyces cerevisiae
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
2.1.1.14
additional information
vitamin-B12 independent enzyme
Candida albicans
2.1.1.14
additional information
vitamin-B12 independent enzyme
Saccharomyces cerevisiae
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
2.1.1.14
0.013
-
L-homocysteine
in 50 mM potassium phosphate (pH 7.2), 50 mM Tris-Cl (pH 7.2) containing 0.1 mM MgSO4, 10 mM dithiothreitol and 2 mM L-homocysteine at 37°C
Saccharomyces cerevisiae
2.1.1.14
0.014
-
L-homocysteine
in 50 mM potassium phosphate (pH 7.2), 50 mM Tris-Cl (pH 7.2) containing 0.1 mM MgSO4, 10 mM dithiothreitol and 2 mM L-homocysteine at 37°C
Candida albicans
2.1.1.14
0.084
-
5-methyltetrahydropteroyl-gamma-glutamyl-gamma-glutamylglutamate
in 50 mM potassium phosphate (pH 7.2), 50 mM Tris-Cl (pH 7.2) containing 0.1 mM MgSO4, 10 mM dithiothreitol and 2 mM L-homocysteine at 37°C
Saccharomyces cerevisiae
2.1.1.14
0.094
-
5-methyltetrahydropteroyl-gamma-glutamyl-gamma-glutamyl-gamma-glutamylglutamate
in 50 mM potassium phosphate (pH 7.2), 50 mM Tris-Cl (pH 7.2) containing 0.1 mM MgSO4, 10 mM dithiothreitol and 2 mM L-homocysteine at 37°C
Saccharomyces cerevisiae
2.1.1.14
0.109
-
5-methyltetrahydropteroyl-gamma-glutamylglutamate
in 50 mM potassium phosphate (pH 7.2), 50 mM Tris-Cl (pH 7.2) containing 0.1 mM MgSO4, 10 mM dithiothreitol and 2 mM L-homocysteine at 37°C
Saccharomyces cerevisiae
2.1.1.14
0.113
-
5-methyltetrahydropteroyl-gamma-glutamylglutamate
in 50 mM potassium phosphate (pH 7.2), 50 mM Tris-Cl (pH 7.2) containing 0.1 mM MgSO4, 10 mM dithiothreitol and 2 mM L-homocysteine at 37°C
Candida albicans
2.1.1.14
0.12
-
5-methyltetrahydropteroyl-gamma-glutamyl-gamma-glutamyl-gamma-glutamylglutamate
in 50 mM potassium phosphate (pH 7.2), 50 mM Tris-Cl (pH 7.2) containing 0.1 mM MgSO4, 10 mM dithiothreitol and 2 mM L-homocysteine at 37°C
Candida albicans
2.1.1.14
0.129
-
5-methyltetrahydropteroyl-gamma-glutamyl-gamma-glutamylglutamate
in 50 mM potassium phosphate (pH 7.2), 50 mM Tris-Cl (pH 7.2) containing 0.1 mM MgSO4, 10 mM dithiothreitol and 2 mM L-homocysteine at 37°C
Candida albicans
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
2.1.1.14
DEAE Sepharose column chromatography
Candida albicans
2.1.1.14
HiTrap heparin column chromatography
Saccharomyces cerevisiae
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
2.1.1.14
additional information
-
42.11 units/mg specific activity for 5-methyltetrahydropteroyl-gamma-glutamyl-gamma-glutamylglutamate after 11.7fold purification, in 50 mM potassium phosphate (pH 7.2), 50 mM Tris-Cl (pH 7.2) containing 0.1 mM MgSO4, 10 mM dithiothreitol and 2 mM L-homocysteine at 37°C
Candida albicans
2.1.1.14
additional information
-
29.3 units/mg specific activity for 5-methyltetrahydropteroyl-gamma-glutamyl-gamma-glutamylglutamate after 5.6fold purification, in 50 mM potassium phosphate (pH 7.2), 50 mM Tris-Cl (pH 7.2) containing 0.1 mM MgSO4, 10 mM dithiothreitol and 2 mM L-homocysteine at 37°C
Saccharomyces cerevisiae
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.1.1.14
5-methyltetrahydropteroyl-gamma-glutamyl-gamma-glutamylglutamate + L-homocysteine
-
671587
Saccharomyces cerevisiae
tetrahydropteroyl-gamma-glutamyl-gamma-glutamylglutamate + L-methionine
-
-
-
?
2.1.1.14
5-methyltetrahydropteroyl-gamma-glutamyl-gamma-glutamylglutamate + L-homocysteine
-
671587
Saccharomyces cerevisiae
tetrahydropteroyl-gamma-glutamyl-gamma-glutamylglutamate + L-methionine
-
-
-
r
2.1.1.14
5-methyltetrahydropteroyl-gamma-glutamyl-gamma-glutamylglutamate + L-homocysteine
-
671587
Candida albicans
tetrahydropteroyl-gamma-glutamyl-gamma-glutamylglutamate + L-methionine
-
-
-
?
2.1.1.14
5-methyltetrahydropteroyl-gamma-glutamyl-gamma-glutamylglutamate + L-homocysteine
-
671587
Candida albicans
tetrahydropteroyl-gamma-glutamyl-gamma-glutamylglutamate + L-methionine
-
-
-
r
2.1.1.14
5-methyltetrahydropteroyl-gamma-glutamylglutamate + L-homocysteine
-
671587
Saccharomyces cerevisiae
tetrahydropteroyl-gamma-glutamylglutamate + L-methionine
-
-
-
?
2.1.1.14
5-methyltetrahydropteroyl-gamma-glutamylglutamate + L-homocysteine
-
671587
Candida albicans
tetrahydropteroyl-gamma-glutamylglutamate + L-methionine
-
-
-
?
2.1.1.14
5-methyltetrahydropteroyltri-L-glutamate + L-homocysteine
-
671587
Saccharomyces cerevisiae
tetrahydropteroyltri-L-glutamate + L-methionine
-
-
-
?
2.1.1.14
5-methyltetrahydropteroyltri-L-glutamate + L-homocysteine
-
671587
Candida albicans
tetrahydropteroyltri-L-glutamate + L-methionine
-
-
-
?
2.1.1.14
additional information
does not use 5-methyltetrahydropteroylglutamate as a substrate
671587
Saccharomyces cerevisiae
?
-
-
-
-
2.1.1.14
additional information
does not use 5-methyltetrahydropteroylglutamate as a substrate
671587
Candida albicans
?
-
-
-
-
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
2.1.1.14
0.283
-
5-methyltetrahydropteroyl-gamma-glutamylglutamate
in 50 mM potassium phosphate (pH 7.2), 50 mM Tris-Cl (pH 7.2) containing 0.1 mM MgSO4, 10 mM dithiothreitol and 2 mM L-homocysteine at 37°C
Saccharomyces cerevisiae
2.1.1.14
0.33
-
L-homocysteine
in 50 mM potassium phosphate (pH 7.2), 50 mM Tris-Cl (pH 7.2) containing 0.1 mM MgSO4, 10 mM dithiothreitol and 2 mM L-homocysteine at 37°C
Saccharomyces cerevisiae
2.1.1.14
0.35
-
5-methyltetrahydropteroyl-gamma-glutamylglutamate
in 50 mM potassium phosphate (pH 7.2), 50 mM Tris-Cl (pH 7.2) containing 0.1 mM MgSO4, 10 mM dithiothreitol and 2 mM L-homocysteine at 37°C
Candida albicans
2.1.1.14
0.4
-
5-methyltetrahydropteroyl-gamma-glutamyl-gamma-glutamylglutamate
in 50 mM potassium phosphate (pH 7.2), 50 mM Tris-Cl (pH 7.2) containing 0.1 mM MgSO4, 10 mM dithiothreitol and 2 mM L-homocysteine at 37°C
Candida albicans
2.1.1.14
0.417
-
5-methyltetrahydropteroyl-gamma-glutamyl-gamma-glutamyl-gamma-glutamylglutamate
in 50 mM potassium phosphate (pH 7.2), 50 mM Tris-Cl (pH 7.2) containing 0.1 mM MgSO4, 10 mM dithiothreitol and 2 mM L-homocysteine at 37°C
Candida albicans
2.1.1.14
0.417
-
L-homocysteine
in 50 mM potassium phosphate (pH 7.2), 50 mM Tris-Cl (pH 7.2) containing 0.1 mM MgSO4, 10 mM dithiothreitol and 2 mM L-homocysteine at 37°C
Candida albicans
2.1.1.14
0.533
-
5-methyltetrahydropteroyl-gamma-glutamyl-gamma-glutamylglutamate
in 50 mM potassium phosphate (pH 7.2), 50 mM Tris-Cl (pH 7.2) containing 0.1 mM MgSO4, 10 mM dithiothreitol and 2 mM L-homocysteine at 37°C
Saccharomyces cerevisiae
2.1.1.14
0.6
-
5-methyltetrahydropteroyl-gamma-glutamyl-gamma-glutamyl-gamma-glutamylglutamate
in 50 mM potassium phosphate (pH 7.2), 50 mM Tris-Cl (pH 7.2) containing 0.1 mM MgSO4, 10 mM dithiothreitol and 2 mM L-homocysteine at 37°C
Saccharomyces cerevisiae