Cloned (Comment) | Organism |
---|---|
recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3) | Ignicoccus islandicus |
Crystallization (Comment) | Organism |
---|---|
purified recombinant His-tagged enzyme, sitting drop vapor diffusion method, mixing of 0.001 ml of 4 mg/ml protein in 50 mM Tris-HCl, pH 7.2, 50 mM NaCl, and 10 mM Tb-Xo4 crystallophore, with 0.001 ml of reservoir solution containing 10% PEG 6000 and 100 mM HEPES, pH 7.0, at 20°C, X-ray diffraction structure determination and analysis, molecular replacement using the structure of Haloferax volcanii MalDH (PDB ID 4BGU) as search template, modeling | Ignicoccus islandicus |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
oxaloacetate | substrate inhibition | Ignicoccus islandicus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | Michaelis-Menten and allosteric sigmoidal kinetics | Ignicoccus islandicus | |
0.38 | - |
oxaloacetate | pH 7.0, 70°C | Ignicoccus islandicus |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
124000 | - |
gel filtration | Ignicoccus islandicus |
133000 | - |
about, sequence calculation | Ignicoccus islandicus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
oxaloacetate + NADH + H+ | Ignicoccus islandicus | - |
L-malate + NAD+ | - |
r | |
oxaloacetate + NADH + H+ | Ignicoccus islandicus DSM 13165 | - |
L-malate + NAD+ | - |
r | |
pyruvate + NADH + H+ | Ignicoccus islandicus | - |
L-lactate + NAD+ | - |
r | |
pyruvate + NADH + H+ | Ignicoccus islandicus DSM 13165 | - |
L-lactate + NAD+ | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Ignicoccus islandicus | A0A0U3FQH7 | - |
- |
Ignicoccus islandicus DSM 13165 | A0A0U3FQH7 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration | Ignicoccus islandicus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | Ignicoccus islandicus MalDH recognizes oxaloacetate as main substrate, but it is also able to use pyruvate | Ignicoccus islandicus | ? | - |
- |
|
additional information | Ignicoccus islandicus MalDH recognizes oxaloacetate as main substrate, but it is also able to use pyruvate | Ignicoccus islandicus DSM 13165 | ? | - |
- |
|
oxaloacetate + NADH + H+ | - |
Ignicoccus islandicus | L-malate + NAD+ | - |
r | |
oxaloacetate + NADH + H+ | best substrate | Ignicoccus islandicus | L-malate + NAD+ | - |
r | |
oxaloacetate + NADH + H+ | - |
Ignicoccus islandicus DSM 13165 | L-malate + NAD+ | - |
r | |
oxaloacetate + NADH + H+ | best substrate | Ignicoccus islandicus DSM 13165 | L-malate + NAD+ | - |
r | |
pyruvate + NADH + H+ | - |
Ignicoccus islandicus | L-lactate + NAD+ | - |
r | |
pyruvate + NADH + H+ | - |
Ignicoccus islandicus DSM 13165 | L-lactate + NAD+ | - |
r |
Subunits | Comment | Organism |
---|---|---|
More | the Ignicoccus islandicus MaDH amino-acid sequence predicts two domains: a NAD(P)-binding Rossmann-fold domain (residues 7 to 138) and a LDH C-terminal-like domain (residues 146 to 308), MalDH quaternary structure analysis and comparison, overview | Ignicoccus islandicus |
tetramer | 4 * 33500, about, sequence calculation | Ignicoccus islandicus |
Synonyms | Comment | Organism |
---|---|---|
LDH-like malate dehydrogenase | - |
Ignicoccus islandicus |
NAD-dependent malate dehydrogenase | - |
Ignicoccus islandicus |
NAD-dependent MalDH | - |
Ignicoccus islandicus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
70 | - |
reduction of oxaloacetate | Ignicoccus islandicus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
8 | - |
pyruvate | pH 7.0, 70°C | Ignicoccus islandicus | |
444 | - |
oxaloacetate | pH 7.0, 70°C | Ignicoccus islandicus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
reduction of oxaloacetate | Ignicoccus islandicus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
additional information | no significant oxaloacetate (OAA) reduction is detected when NADPH was used as coenzyme at 70°C. the presence of an aspartic residue at position 54 of the alignment suggests that Ignicoccus islandicus MalDH would use NADH instead of NADPH, and the lack of NADPH recognition confirmes it. In contrast, those characterized as NADPH-dependent MalDHs have a glycine residue at this position. This is because the smaller size of glycine allows accommodating the additional phosphate of NADPH | Ignicoccus islandicus | |
NAD+ | - |
Ignicoccus islandicus | |
NADH | - |
Ignicoccus islandicus |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.24 | - |
oxaloacetate | pH 7.0, 70°C | Ignicoccus islandicus |
General Information | Comment | Organism |
---|---|---|
evolution | the enzyme belongs to the MalDH/LDH superfamily, which is divided into several phylogenetically related groups, lactate dehydrogenases (LDHs) and malate dehydrogenases (MalDHs) belong to a wide group of 2-oxoacid:NAD(P)-dependent dehydrogenases that catalyze the reversible conversion of 2-hydroxyacids to the corresponding 2-oxoacids, evolutionary history of the LDHs and MalDHs, overview. The enzyme structure belongs to the NAD(P)-binding Rossmann-like domain CATH superfamily | Ignicoccus islandicus |
metabolism | when the competent catalytic state is reached, LDH catalyzes the direct transfer of a hydride ion from the pro-R face of NADH to the C2 carbon of pyruvate to produce lactate, whereas MalDH transforms oxaloacetate into malate | Ignicoccus islandicus |
additional information | MalDH is an enzyme with intermediate properties between allosteric LDHs and non-allosteric tetrameric MalDHs. The catalytic residue is histidine H195. The structure of Ignicoccus islandicus MalDH resembles that of canonical LDHs. The amino acid at position 102 is considered as the most important substrate-discriminating residue between LDHs and MalDHs. Structure-function analysis and comparisons, overview | Ignicoccus islandicus |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1168.4 | - |
oxaloacetate | pH 7.0, 70°C | Ignicoccus islandicus |