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Literature summary for 1.1.1.37 extracted from

  • Rozova, O.N.; Khmelenina, V.N.; Bocharova, K.A.; Mustakhimov, I.I.; Trotsenko, Y.A.
    Role of NAD+-dependent malate dehydrogenase in the metabolism of Methylomicrobium alcaliphilum 20Z and Methylosinus trichosporium OB3b (2015), Microorganisms, 3, 47-59.
    View publication on PubMedView publication on EuropePMC

Activating Compound

Activating Compound Comment Organism Structure
additional information the thermostable enzyme is not affected by metal ions (CuCl2, MgCl2, MnCl2, CoCl2, BaCl2, ZnCl2 or CaCl2 and NaCl or KCl) or various organic metabolites (pyruvate, phosphoenolpyruvate, ATP, ADP, AMP, glucose-1-phospate, fructose-1-phosphate, fructose-1,6-bisphosphate, fructose-6-phosphate, ribose-1-phosphate, ribose-5-phosphate (all 5 mM), glycerate, lactate, 2-oxoglutarate, citrate, serine, diphosphate (all 1 mM) or phosphate (10 mM)) Methylotuvimicrobium alcaliphilum
additional information the thermostable enzyme is not affected by metal ions (CuCl2, MgCl2, MnCl2, CoCl2, BaCl2, ZnCl2 or CaCl2 and NaCl or KCl) or various organic metabolites (Pyruvate, phosphoenolpyruvate, ATP, ADP, AMP, glucose-1-phospate, fructose-1-phosphate, fructose-1,6-bisphosphate, fructose-6-phosphate, ribose-1-phosphate, ribose-5-phosphate (all 5 mM), glycerate, lactate, 2-oxoglutarate, citrate, serine, pyrophosphate (all 1 mM) or ?H2PO4 (10 mM)) Methylosinus trichosporium

Cloned(Commentary)

Cloned (Comment) Organism
gene mdh, recombinant expression of His6-tagged enzyme in Escherichia coli strain BL21(DE3) Methylosinus trichosporium
gene mdh, recombinant expression of His6-tagged enzyme in Escherichia coli strain BL21(DE3) Methylotuvimicrobium alcaliphilum

Inhibitors

Inhibitors Comment Organism Structure
additional information the thermostable enzyme is not affected by metal ions (CuCl2, MgCl2, MnCl2, CoCl2, BaCl2, ZnCl2 or CaCl2 and NaCl or KCl) or various organic metabolites (pyruvate, phosphoenolpyruvate, ATP, ADP, AMP, glucose-1-phospate, fructose-1-phosphate, fructose-1,6-bisphosphate, fructose-6-phosphate, ribose-1-phosphate, ribose-5-phosphate (all 5 mM), glycerate, lactate, 2-oxoglutarate, citrate, serine, diphosphate (all 1 mM) or phosphate (10 mM)) Methylosinus trichosporium
additional information the thermostable enzyme is not affected by metal ions (CuCl2, MgCl2, MnCl2, CoCl2, BaCl2, ZnCl2 or CaCl2 and NaCl or KCl) or various organic metabolites (Pyruvate, phosphoenolpyruvate, ATP, ADP, AMP, glucose-1-phospate, fructose-1-phosphate, fructose-1,6-bisphosphate, fructose-6-phosphate, ribose-1-phosphate, ribose-5-phosphate (all 5 mM), glycerate, lactate, 2-oxoglutarate, citrate, serine, pyrophosphate (all 1 mM) or ?H2PO4 (10 mM)) Methylotuvimicrobium alcaliphilum

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information Michaelis-Menten kinetics Methylosinus trichosporium
additional information
-
additional information Michaelis-Menten kinetics Methylotuvimicrobium alcaliphilum
0.0252
-
NADH recombinant enzyme, pH 10.0, 30°C Methylotuvimicrobium alcaliphilum
0.0376
-
NADH recombinant enzyme, pH 9.5, 30°C Methylosinus trichosporium
0.059
-
oxaloacetate recombinant enzyme, pH 9.5, 30°C Methylosinus trichosporium
0.11
-
(S)-malate recombinant enzyme, pH 10.0, 30°C Methylotuvimicrobium alcaliphilum
0.33
-
NAD+ recombinant enzyme, pH 9.5, 30°C Methylosinus trichosporium
0.34
-
oxaloacetate recombinant enzyme, pH 10.0, 30°C Methylotuvimicrobium alcaliphilum
0.45
-
NAD+ recombinant enzyme, pH 10.0, 30°C Methylotuvimicrobium alcaliphilum
1.28
-
(S)-malate recombinant enzyme, pH 9.5, 30°C Methylosinus trichosporium

Metals/Ions

Metals/Ions Comment Organism Structure
additional information the thermostable enzyme is not affected by metal ions (CuCl2, MgCl2, MnCl2, CoCl2, BaCl2, ZnCl2 or CaCl2 and NaCl or KCl) or various organic metabolites (pyruvate, phosphoenolpyruvate, ATP, ADP, AMP, glucose-1-phospate, fructose-1-phosphate, fructose-1,6-bisphosphate, fructose-6-phosphate, ribose-1-phosphate, ribose-5-phosphate (all 5 mM), glycerate, lactate, 2-oxoglutarate, citrate, serine, diphosphate (all 1 mM) or phosphate (10 mM)) Methylosinus trichosporium
additional information the thermostable enzyme is not affected by metal ions (CuCl2, MgCl2, MnCl2, CoCl2, BaCl2, ZnCl2 or CaCl2 and NaCl or KCl) or various organic metabolites (pyruvate, phosphoenolpyruvate, ATP, ADP, AMP, glucose-1-phospate, fructose-1-phosphate, fructose-1,6-bisphosphate, fructose-6-phosphate, ribose-1-phosphate, ribose-5-phosphate (all 5 mM), glycerate, lactate, 2-oxoglutarate, citrate, serine, diphosphate (all 1 mM) or phosphate (10 mM)) Methylotuvimicrobium alcaliphilum

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
33200
-
-
Methylosinus trichosporium
35000
-
-
Methylosinus trichosporium
35000
-
-
Methylotuvimicrobium alcaliphilum
35800
-
-
Methylotuvimicrobium alcaliphilum
70000
-
recombinant enzyme, gel filtration Methylotuvimicrobium alcaliphilum
140000
-
recombinant enzyme, gel filtration Methylosinus trichosporium

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
(S)-malate + NAD+ Methylosinus trichosporium
-
oxaloacetate + NADH + H+
-
r
(S)-malate + NAD+ Methylotuvimicrobium alcaliphilum
-
oxaloacetate + NADH + H+
-
r
(S)-malate + NAD+ Methylotuvimicrobium alcaliphilum 20Z
-
oxaloacetate + NADH + H+
-
r
oxaloacetate + NADH + H+ Methylosinus trichosporium
-
(S)-malate + NAD+
-
r
oxaloacetate + NADH + H+ Methylotuvimicrobium alcaliphilum
-
(S)-malate + NAD+
-
r
oxaloacetate + NADH + H+ Methylotuvimicrobium alcaliphilum 20Z
-
(S)-malate + NAD+
-
r

Organism

Organism UniProt Comment Textmining
Methylosinus trichosporium
-
-
-
Methylotuvimicrobium alcaliphilum G4T3S9
-
-
Methylotuvimicrobium alcaliphilum 20Z G4T3S9
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography Methylosinus trichosporium
recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography Methylotuvimicrobium alcaliphilum

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
15.05
-
purified recombinant enzyme, oxaloacetate reduction, pH 10.0, 30°C Methylotuvimicrobium alcaliphilum
20.75
-
purified recombinant enzyme, malate oxidation, pH 10.0, 30°C Methylotuvimicrobium alcaliphilum
78.2
-
purified recombinant enzyme, malate oxidation, pH 9.5, 30°C Methylosinus trichosporium
187.8
-
purified recombinant enzyme, oxaloacetate reduction, pH 9.5, 30°C Methylosinus trichosporium

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(S)-malate + NAD+
-
Methylosinus trichosporium oxaloacetate + NADH + H+
-
r
(S)-malate + NAD+
-
Methylotuvimicrobium alcaliphilum oxaloacetate + NADH + H+
-
r
(S)-malate + NAD+
-
Methylotuvimicrobium alcaliphilum 20Z oxaloacetate + NADH + H+
-
r
additional information the enzyme displays nearly equal reductive (malate formation) and oxidative (oxaloacetate formation) activities and higher affinity to malate than to oxaloacetate Methylotuvimicrobium alcaliphilum ?
-
?
additional information the enzyme is 2fold more active in the reaction of oxaloacetate reduction compared to malate oxidation and exhibiting higher affinity to oxaloacetate than to malate Methylosinus trichosporium ?
-
?
additional information the enzyme displays nearly equal reductive (malate formation) and oxidative (oxaloacetate formation) activities and higher affinity to malate than to oxaloacetate Methylotuvimicrobium alcaliphilum 20Z ?
-
?
oxaloacetate + NADH + H+
-
Methylosinus trichosporium (S)-malate + NAD+
-
r
oxaloacetate + NADH + H+
-
Methylotuvimicrobium alcaliphilum (S)-malate + NAD+
-
r
oxaloacetate + NADH + H+
-
Methylotuvimicrobium alcaliphilum 20Z (S)-malate + NAD+
-
r

Subunits

Subunits Comment Organism
homodimer 2 * 35000, recombinant enzyme, SDS-PAGE, 2 * 35800, about, sequence calculation Methylotuvimicrobium alcaliphilum
homotetramer 4 * 35000, recombinant enzyme, SDS-PAGE, 4 * 33200, about, sequence calculation Methylosinus trichosporium

Synonyms

Synonyms Comment Organism
A8B73_11505
-
Methylosinus trichosporium
L-malate: NAD oxidoreductase
-
Methylosinus trichosporium
L-malate: NAD oxidoreductase
-
Methylotuvimicrobium alcaliphilum
MDH
-
Methylosinus trichosporium
MDH
-
Methylotuvimicrobium alcaliphilum
NAD+-dependent malate dehydrogenase
-
Methylosinus trichosporium
NAD+-dependent malate dehydrogenase
-
Methylotuvimicrobium alcaliphilum

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
60
-
-
Methylosinus trichosporium
60 65
-
Methylotuvimicrobium alcaliphilum

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
30 60 the activity of the methanotroph's enzyme increases up to 60°? and then gradually decreases Methylosinus trichosporium
30 60 the activity of the methanotroph's enzyme increases up to 60°? and then gradually decreases Methylotuvimicrobium alcaliphilum

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
30 70 the enzyme is active in a broader temperature range. It retains full activity after 18 h exposure at 30-60°C, exhibiting a 1 h half-life time at 70°C Methylosinus trichosporium
30 60 the purified recombinant His6-tagged MDH from strain 20Z fully retains activity after 18 h exposure at 30-40°C, and has a 10% residual activity after 18 h exposure at 50°C. The enzyme loses 50% of its activity after 10 min heating at 60°C Methylotuvimicrobium alcaliphilum

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
516
-
oxaloacetate recombinant enzyme, pH 9.5, 30°C Methylosinus trichosporium
621
-
(S)-malate recombinant enzyme, pH 9.5, 30°C Methylotuvimicrobium alcaliphilum
870
-
oxaloacetate recombinant enzyme, pH 9.5, 30°C Methylotuvimicrobium alcaliphilum
1141
-
(S)-malate recombinant enzyme, pH 9.5, 30°C Methylosinus trichosporium

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
9.5
-
-
Methylosinus trichosporium
10
-
-
Methylotuvimicrobium alcaliphilum

pH Range

pH Minimum pH Maximum Comment Organism
7.5 10 activity range, recombinant enzyme Methylosinus trichosporium
7.5 10 activity range, recombinant enzyme Methylotuvimicrobium alcaliphilum

Cofactor

Cofactor Comment Organism Structure
NAD+
-
Methylosinus trichosporium
NAD+
-
Methylotuvimicrobium alcaliphilum
NADH
-
Methylosinus trichosporium
NADH
-
Methylotuvimicrobium alcaliphilum

General Information

General Information Comment Organism
metabolism the enzyme plays crucial roles in many metabolic pathways, including the tricarboxylic acid (TCA) cycle, energy generation and the formation of metabolites for biosynthesis Methylosinus trichosporium
metabolism the enzyme plays crucial roles in many metabolic pathways, including the tricarboxylic acid (TCA) cycle, energy generation and the formation of metabolites for biosynthesis Methylotuvimicrobium alcaliphilum

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
403
-
(S)-malate recombinant enzyme, pH 9.5, 30°C Methylosinus trichosporium
2558
-
oxaloacetate recombinant enzyme, pH 9.5, 30°C Methylotuvimicrobium alcaliphilum
5648
-
(S)-malate recombinant enzyme, pH 9.5, 30°C Methylotuvimicrobium alcaliphilum
19340
-
oxaloacetate recombinant enzyme, pH 9.5, 30°C Methylosinus trichosporium