1.1.1.37: malate dehydrogenase

This is an abbreviated version, for detailed information about malate dehydrogenase, go to the full flat file.

Reaction

(S)-malate
+
NAD+
=
oxaloacetate
+
NADH
+
H+

Synonyms

(R)-2-hydroxyacid dehydrogenase, (S)-malate dehydrogenase, CaMDH, cMDH, cMDH -S, cMDH-L, cyMDH, cytosolic malate dehydrogenase, cytosolic MDH, cytosolic NAD-dependent malate dehydrogenase, halophilic malate dehydrogenase, HmMalDH, L-malate dehydrogenase, L-malate-NAD oxidoreductase, L-malate: NAD oxidoreductase, L-malate: NAD+ oxidoreductase, L-malate:NAD-oxidoreductase, L-MDH, m-MDH, malate (NAD) dehydrogenase, malate dehydrogenase, malate dehydrogenase (NAD), malate dehydrogenase 1, malate dehydrogenase 2, malate: NAD oxidoreductase, MalDH, malic acid dehydrogenase, malic dehydrogenase, mbNAD-MDH, MDH, MDH A, MDH B1, MDH B2, Mdh1, MDH2, Mdh2a, Mdh2b, mitochondrial malate dehydrogenase, mitochondrial MDH, mMDH, mNAD-MDH, More, NAD+-dependent malate dehydrogenase, NAD+-dependent MDH, NAD+-MDH enzymes, NAD-dependent malate dehydrogenase, NAD-dependent malic dehydrogenase, NAD-dependent MDH, NAD-L-malate dehydrogenase, NAD-linked malate dehydrogenase, NAD-malate dehydrogenase, NAD-malic dehydrogenase, NAD-MDH, NAD-specific malate dehydrogenase, Pcal_1699, peroxisomal NAD+-malate dehydrogenase 1, peroxisomal NAD+-malate dehydrogenase 2, PMDH, PMDH1, PMDH2, regulatory subunit of nucleic acid-conducting channel, s-MDH, SrMalDH, TaMDH, VEG69, Vegetative protein 69, YlMdh2p, [LDH-like] MDH

ECTree

     1 Oxidoreductases
         1.1 Acting on the CH-OH group of donors
             1.1.1 With NAD+ or NADP+ as acceptor
                1.1.1.37 malate dehydrogenase

Engineering

Engineering on EC 1.1.1.37 - malate dehydrogenase

Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
E165K
increase in temperature-optimum to 65°C, compared to 60°C for the wild-type enzyme. Maximal specific activity at temperature optimum is increased by about 30% compared to wild-type enzyme. Melting temperature at pH 4.4 is by 4.6°C, melting temperature at pH 6.0 is increased by 12.0°C and melting temperature at pH 7.5 is increased by 23.9°C compared to wild-type enzyme, mutation stabilizes to such an extent that disruption of the inter-dimer electrostatic network around residue 165 no longer limits kinetic thermal stability
E165Q
increase in temperature-optimum to 65°C, compared to 60°C for the wild-type enzyme. Maximal specific activity at temperature optimum is increased by about 30% compared to wild-type enzyme. Melting temperature at pH 4.4 is by 5.4°C, melting temperature at pH 6.0 is increased by 11.2°C and melting temperature at pH 7.5 is increased by 23.6°C compared to wild-type enzyme, mutation stabilizes to such an extent that disruption of the inter-dimer electrostatic network around residue 165 no longer limits kinetic thermal stability
F144I
site-directed mutagenesis, inactive mutant
G210A
site-directed mutagenesis, the mutant shows 30% reduced activity compared to the wild-type enzyme
G210A/V214I
site-directed mutagenesis, the double mutant shows a 2.2fold increase in lacatate dehydrogenase activity compared to the wild-type enzyme
I12V/R81Q/M85E/G210A/V214I
construction of a pentamutant by site-directed mutagenesis, whose substrate specificity is switched from malate dehydrogenase to that of lactate dehydrogenase, EC 1.1.1.27, the mutant shows highly reduced activity compared to the wild-type enzyme, overview
N122D
site-directed mutagenesis, inactive mutant
R81Q
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
V214I
site-directed mutagenesis, the mutant's activity is not affected
E243R
-
mutation does not affect enzyme activity. The mutant enzyme is more halophilic than the wild-type protein. It is more sensitive to temperature and requires significantly higher concentrations of NaCl or KCl for equivalent stability
E267R
-
the numbering is not equivalent to the numbering of UniProt. The E267R mutation points into a central ordered water cavity, disrupting protein-solvent interactions. The mutant enzyme requires higher concentrations of the solvent salt for stability similar to that of the wild type
K205A
-
site-directed mutagenesis, the oligomeric state of the mutant changes with the nature of the anion bound, the mutant is dimeric or tetrameric, overview
R100Q
-
the mutant enzyme has considerably higher specificity for pyruvate than for oxaloacetate. Whereas the Km value for pyruvate is increased about 2fold, that for oxaloacetate increases 30fold. The R100Q mutant is not subjected to substrate inhibition, at least not at substrate concentrations up to 30 mM, and the highest kcat value is obtained at the lowest salt concentration used (0.15 M NaCl)
R207S/R292S
E243R
-
mutation does not affect enzyme activity. The mutant enzyme is more halophilic than the wild-type protein. It is more sensitive to temperature and requires significantly higher concentrations of NaCl or KCl for equivalent stability
-
E267R
-
the numbering is not equivalent to the numbering of UniProt. The E267R mutation points into a central ordered water cavity, disrupting protein-solvent interactions. The mutant enzyme requires higher concentrations of the solvent salt for stability similar to that of the wild type
-
R100Q
-
the mutant enzyme has considerably higher specificity for pyruvate than for oxaloacetate. Whereas the Km value for pyruvate is increased about 2fold, that for oxaloacetate increases 30fold. The R100Q mutant is not subjected to substrate inhibition, at least not at substrate concentrations up to 30 mM, and the highest kcat value is obtained at the lowest salt concentration used (0.15 M NaCl)
-
R207S/R292S
-
the numbering is not equivalent to the numbering of UniProt. The active tetrameric mutant enzyme R207S/R292S dissociates under certain conditions to active dimers and under other conditions to inactive dimers. These dimers further dissociate into folded monomers which eventually unfold. The mutant enzyme requires higher salt concentrations than the wild type for stability. Thermal inactivation starts at 35°C, whereas the wild type is stable up to 60°C. At 4 M NaCl (pH 8) the kinetics of unfolding of the mutant is measured by following the fluorescence emission during incubation at various temperatures. The process is biphasic between 35 and 48 °C, while the thermal deactivation kinetics of the wild type protein is first-order; the numbering is not equivalent to the numbering of UniProt. The mutations, located at the dimer-dimer interface, disrupt two inter-dimeric salt bridge clusters that are essential for wild-type tetramer stabilisation. Association of active dimers into tetramers is weakened
-
H187Y
-
catalytically inactive
H229Q
mutant enzyme is less resistant to heat denaturation than the wild-type enzyme
Q229H
mutant enzyme is more resistant to heat denaturation than the wild-type enzyme
H229Q
-
mutant enzyme is less resistant to heat denaturation than the wild-type enzyme
-
Q229H
-
mutant enzyme is more resistant to heat denaturation than the wild-type enzyme
-
V114H
decrease in thermal stability, decrease in KM-value and kcat
V144N
decrease in thermal stability
R183a
-
dimeric mutant with almost wild type activity
R214G
-
dimeric mutant with almost wild type activity
additional information