BRENDA - Enzyme Database

Enzymatic function of loop movement in enolase: preparation and some properties of H159N, H159A, H159F, and N207A enolases

Brewer, J.M.; Glover, C.V.; Holland, M.J.; Lebioda, L.; J. Protein Chem. 22, 353-361 (2003)

Data extracted from this reference:

Engineering
EC Number
Amino acid exchange
Commentary
Organism
4.2.1.11
H159A
less than 1% of the activity compared to the wild type
Saccharomyces cerevisiae
4.2.1.11
H159F
less than 1% of the activity compared to the wild type
Saccharomyces cerevisiae
4.2.1.11
H159N
less than 1% of the activity compared to the wild type
Saccharomyces cerevisiae
4.2.1.11
N207A
50% of the activity compared to the wild type
Saccharomyces cerevisiae
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
4.2.1.11
Mg2+
inhibitor above 1 mM, N207A, H159A, H159N and H159F mutants are stimulated at this concentration
Saccharomyces cerevisiae
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
4.2.1.11
additional information
-
2-phospho-D-glycerate
Km ranges between 0.047 mM and 0.130 mM between pH 6.0 and pH 8.6
Saccharomyces cerevisiae
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
4.2.1.11
96000
-
gel filtration, mutant N207A enzyme; gel filtration, wild type enzyme
Saccharomyces cerevisiae
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
4.2.1.11
Saccharomyces cerevisiae
-
isozyme 1
-
Purification (Commentary)
EC Number
Commentary
Organism
4.2.1.11
wild type and several mutant recombinant enzymes using chromatography on Sepharose
Saccharomyces cerevisiae
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
4.2.1.11
1.1
-
H159N mutant, pH 7.8, 21ºC
Saccharomyces cerevisiae
4.2.1.11
1.9
-
H159F mutant, pH 7.8, 21ºC
Saccharomyces cerevisiae
4.2.1.11
3.3
-
H159A mutant, pH 7.8, 21ºC
Saccharomyces cerevisiae
4.2.1.11
248
-
N207A mutant, pH 7.8, 21ºC
Saccharomyces cerevisiae
4.2.1.11
442
-
wild type, pH 7.8, 21ºC
Saccharomyces cerevisiae
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
4.2.1.11
2-phospho-D-glycerate
-
653043
Saccharomyces cerevisiae
phosphoenolpyruvate
-
653043
Saccharomyces cerevisiae
r
4.2.1.11
D-tartronate semialdehyde-2-phosphate
substrate analogue that changes its spectrum while bound to the enzyme
653043
Saccharomyces cerevisiae
?
-
653043
Saccharomyces cerevisiae
?
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
4.2.1.11
H159A
less than 1% of the activity compared to the wild type
Saccharomyces cerevisiae
4.2.1.11
H159F
less than 1% of the activity compared to the wild type
Saccharomyces cerevisiae
4.2.1.11
H159N
less than 1% of the activity compared to the wild type
Saccharomyces cerevisiae
4.2.1.11
N207A
50% of the activity compared to the wild type
Saccharomyces cerevisiae
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
4.2.1.11
Mg2+
inhibitor above 1 mM, N207A, H159A, H159N and H159F mutants are stimulated at this concentration
Saccharomyces cerevisiae
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
4.2.1.11
additional information
-
2-phospho-D-glycerate
Km ranges between 0.047 mM and 0.130 mM between pH 6.0 and pH 8.6
Saccharomyces cerevisiae
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
4.2.1.11
96000
-
gel filtration, mutant N207A enzyme; gel filtration, wild type enzyme
Saccharomyces cerevisiae
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
4.2.1.11
wild type and several mutant recombinant enzymes using chromatography on Sepharose
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
4.2.1.11
1.1
-
H159N mutant, pH 7.8, 21ºC
Saccharomyces cerevisiae
4.2.1.11
1.9
-
H159F mutant, pH 7.8, 21ºC
Saccharomyces cerevisiae
4.2.1.11
3.3
-
H159A mutant, pH 7.8, 21ºC
Saccharomyces cerevisiae
4.2.1.11
248
-
N207A mutant, pH 7.8, 21ºC
Saccharomyces cerevisiae
4.2.1.11
442
-
wild type, pH 7.8, 21ºC
Saccharomyces cerevisiae
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
4.2.1.11
2-phospho-D-glycerate
-
653043
Saccharomyces cerevisiae
phosphoenolpyruvate
-
653043
Saccharomyces cerevisiae
r
4.2.1.11
D-tartronate semialdehyde-2-phosphate
substrate analogue that changes its spectrum while bound to the enzyme
653043
Saccharomyces cerevisiae
?
-
653043
Saccharomyces cerevisiae
?