BRENDA - Enzyme Database

Studies on regulatory functions of malic enzymes II. Purification and molecular properties of nicotinamide adenine dinucleotide-linked malic enzyme from Escherichia coli

Yamaguchi M.; Tokushige, M.; Katsuki, H.; J. Biochem. 73, 169-180 (1973)

Data extracted from this reference:

Activating Compound
EC Number
Activating Compound
Commentary
Organism
Structure
1.1.1.38
L-aspartate
-
Escherichia coli W
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
1.1.1.38
1,10-phenanthroline
0.025 mM, 70% activity lost after 5 min
Escherichia coli W
1.1.1.38
CoA
-
Escherichia coli W
1.1.1.38
Cu2+
0.025 mM, 70% activity lost after 5 min
Escherichia coli W
1.1.1.38
D-malate
competitive inhibition
Escherichia coli W
1.1.1.38
malonate
competitive inhibition
Escherichia coli W
1.1.1.38
oxaloacetate
50% inhibition with 0.09 mM
Escherichia coli W
1.1.1.38
p-(chloromercuri)benzoate
0.0025 mM, 80% inhibition within 2 min
Escherichia coli W
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.1.1.38
0.055
-
NAD+
-
Escherichia coli W
1.1.1.38
0.32
-
Mn2+
-
Escherichia coli W
1.1.1.38
0.4
-
L-malate
-
Escherichia coli W
1.1.1.38
2.3
-
NADP+
-
Escherichia coli W
1.1.1.38
5
-
oxaloacetate
-
Escherichia coli W
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
1.1.1.38
Co2+
-
Escherichia coli W
1.1.1.38
Mg2+
-
Escherichia coli W
1.1.1.38
Mn2+
-
Escherichia coli W
1.1.1.38
Ni2+
requirement for divalent cation
Escherichia coli W
1.1.1.38
Zn2+
requirement for divalent cation
Escherichia coli W
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
1.1.1.38
198000
-
low speed sedimentation equilibrium
Escherichia coli W
1.1.1.38
203000
-
sucrose density gradient centrifugation
Escherichia coli W
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.1.1.38
(S)-malate + NAD+
Escherichia coli W
-
CO2 + pyruvate + NADH
-
Escherichia coli W
-
1.1.1.38
Oxaloacetate
Escherichia coli W
-
CO2 + pyruvate
-
Escherichia coli W
ir
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
1.1.1.38
Escherichia coli W
-
-
-
Oxidation Stability
EC Number
Oxidation Stability
Organism
1.1.1.38
enzyme is reversibly inhibited by air oxidation, activity is completely restored in 30 min by the addition of 2 mM dithiotreitol
Escherichia coli W
Purification (Commentary)
EC Number
Commentary
Organism
1.1.1.38
streptomycin, ammonium sulfate, heat treatment, acid treatment, calcium phosphate gel, DEAE-sephadex, hydroxylapatite, Sepharose 6B
Escherichia coli W
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
1.1.1.38
177
-
-
Escherichia coli W
Storage Stability
EC Number
Storage Stability
Organism
1.1.1.38
4°C, 50 mM Tris-HCl, pH 7.4, 1 mM EDTA, 10 mM 2-mercaptoethanol, 10 mM MgCl2, 5 mM L-aspartate, 2 months, 10% activity lost
Escherichia coli W
1.1.1.38
4°C, 50 mM Tris-HCl, pH 7.4, 1 mM EDTA, 10 mM 2-mercaptoethanol, 2 months, 15% activity lost
Escherichia coli W
1.1.1.38
4°C, 50 mM Tris-HCl, pH 7.4, 1 month, 90% activity lost
Escherichia coli W
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.1.1.38
(S)-malate + NAD+
-
286697
Escherichia coli W
CO2 + pyruvate + NADH
-
286697
Escherichia coli W
-
1.1.1.38
(S)-malate + NAD+
-
286697
Escherichia coli W
CO2 + pyruvate + NADH
-
286697
Escherichia coli W
r
1.1.1.38
(S)-malate + NADP+
-
286697
Escherichia coli W
CO2 + pyruvate + NADPH
-
-
-
-
1.1.1.38
Oxaloacetate
-
286697
Escherichia coli W
CO2 + pyruvate
-
286697
Escherichia coli W
ir
1.1.1.38
Oxaloacetate
NAD+ inhibits oxaloacetate decarboxylation activity
286697
Escherichia coli W
CO2 + pyruvate
-
286697
Escherichia coli W
ir
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.1.1.38
7.2
-
with increasing substrate concentrations pH optimum is shifted towards higher pH values
Escherichia coli W
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
1.1.1.38
NAD+
-
Escherichia coli W
1.1.1.38
NADP+
2% activity
Escherichia coli W
Activating Compound (protein specific)
EC Number
Activating Compound
Commentary
Organism
Structure
1.1.1.38
L-aspartate
-
Escherichia coli W
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
1.1.1.38
NAD+
-
Escherichia coli W
1.1.1.38
NADP+
2% activity
Escherichia coli W
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
1.1.1.38
1,10-phenanthroline
0.025 mM, 70% activity lost after 5 min
Escherichia coli W
1.1.1.38
CoA
-
Escherichia coli W
1.1.1.38
Cu2+
0.025 mM, 70% activity lost after 5 min
Escherichia coli W
1.1.1.38
D-malate
competitive inhibition
Escherichia coli W
1.1.1.38
malonate
competitive inhibition
Escherichia coli W
1.1.1.38
oxaloacetate
50% inhibition with 0.09 mM
Escherichia coli W
1.1.1.38
p-(chloromercuri)benzoate
0.0025 mM, 80% inhibition within 2 min
Escherichia coli W
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.1.1.38
0.055
-
NAD+
-
Escherichia coli W
1.1.1.38
0.32
-
Mn2+
-
Escherichia coli W
1.1.1.38
0.4
-
L-malate
-
Escherichia coli W
1.1.1.38
2.3
-
NADP+
-
Escherichia coli W
1.1.1.38
5
-
oxaloacetate
-
Escherichia coli W
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
1.1.1.38
Co2+
-
Escherichia coli W
1.1.1.38
Mg2+
-
Escherichia coli W
1.1.1.38
Mn2+
-
Escherichia coli W
1.1.1.38
Ni2+
requirement for divalent cation
Escherichia coli W
1.1.1.38
Zn2+
requirement for divalent cation
Escherichia coli W
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
1.1.1.38
198000
-
low speed sedimentation equilibrium
Escherichia coli W
1.1.1.38
203000
-
sucrose density gradient centrifugation
Escherichia coli W
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.1.1.38
(S)-malate + NAD+
Escherichia coli W
-
CO2 + pyruvate + NADH
-
Escherichia coli W
-
1.1.1.38
Oxaloacetate
Escherichia coli W
-
CO2 + pyruvate
-
Escherichia coli W
ir
Oxidation Stability (protein specific)
EC Number
Oxidation Stability
Organism
1.1.1.38
enzyme is reversibly inhibited by air oxidation, activity is completely restored in 30 min by the addition of 2 mM dithiotreitol
Escherichia coli W
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
1.1.1.38
streptomycin, ammonium sulfate, heat treatment, acid treatment, calcium phosphate gel, DEAE-sephadex, hydroxylapatite, Sepharose 6B
Escherichia coli W
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
1.1.1.38
177
-
-
Escherichia coli W
Storage Stability (protein specific)
EC Number
Storage Stability
Organism
1.1.1.38
4°C, 50 mM Tris-HCl, pH 7.4, 1 mM EDTA, 10 mM 2-mercaptoethanol, 10 mM MgCl2, 5 mM L-aspartate, 2 months, 10% activity lost
Escherichia coli W
1.1.1.38
4°C, 50 mM Tris-HCl, pH 7.4, 1 mM EDTA, 10 mM 2-mercaptoethanol, 2 months, 15% activity lost
Escherichia coli W
1.1.1.38
4°C, 50 mM Tris-HCl, pH 7.4, 1 month, 90% activity lost
Escherichia coli W
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.1.1.38
(S)-malate + NAD+
-
286697
Escherichia coli W
CO2 + pyruvate + NADH
-
286697
Escherichia coli W
-
1.1.1.38
(S)-malate + NAD+
-
286697
Escherichia coli W
CO2 + pyruvate + NADH
-
286697
Escherichia coli W
r
1.1.1.38
(S)-malate + NADP+
-
286697
Escherichia coli W
CO2 + pyruvate + NADPH
-
-
-
-
1.1.1.38
Oxaloacetate
-
286697
Escherichia coli W
CO2 + pyruvate
-
286697
Escherichia coli W
ir
1.1.1.38
Oxaloacetate
NAD+ inhibits oxaloacetate decarboxylation activity
286697
Escherichia coli W
CO2 + pyruvate
-
286697
Escherichia coli W
ir
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.1.1.38
7.2
-
with increasing substrate concentrations pH optimum is shifted towards higher pH values
Escherichia coli W