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Literature summary extracted from

  • Oikawa, T.; Yamamoto, N.; Shimoke, K.; Uesato, S.; Ikeuchi, T.; Fujioka, T.
    Purification, characterization, and overexpression of psychrophilic and thermolabile malate dehydrogenase of a novel antarctic psychrotolerant, Flavobacterium frigidimaris KUC-1 (2005), Biosci. Biotechnol. Biochem., 69, 2146-2154.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.37 expression in Escherichia coli Flavobacterium frigidimaris

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.1.1.37 BaCl2 1 mM, 24.9% inhibition Flavobacterium frigidimaris
1.1.1.37 CaCl2 1 mM, 29.1% inhibition Flavobacterium frigidimaris
1.1.1.37 CoCl2 1 mM, 39% inhibition Flavobacterium frigidimaris
1.1.1.37 CuCl2 1 mM, 89.8% inhibition Flavobacterium frigidimaris
1.1.1.37 EDTA 1 mM, 52.2% inhibition Flavobacterium frigidimaris
1.1.1.37 HgCl2 1 mM, complete inhibition Flavobacterium frigidimaris
1.1.1.37 hydroxylamine 1 mM, 57.1% inhibition Flavobacterium frigidimaris
1.1.1.37 iodoacetate 1 mM, 93.6% inhibition Flavobacterium frigidimaris
1.1.1.37 MgCl2 1 mM,34.9% inhibition Flavobacterium frigidimaris
1.1.1.37 MnCl2 1 mM, 28.6% inhibition Flavobacterium frigidimaris
1.1.1.37 NEM 1 mM, complete inhibition Flavobacterium frigidimaris
1.1.1.37 NiCl2 1 mM, 36.8% inhibition Flavobacterium frigidimaris
1.1.1.37 Semicarbazide 1 mM, 61.3% inhibition Flavobacterium frigidimaris
1.1.1.37 ZnCl2 1 mM, 72.6% inhibition Flavobacterium frigidimaris
1.1.1.299 CuCl2 1 mM, 10% residual activity Flavobacterium frigidimaris
1.1.1.299 HgCl2 1 mM, no residual activity Flavobacterium frigidimaris
1.1.1.299 iodoacetate 1 mM, 16.4% residual activity Flavobacterium frigidimaris
1.1.1.299 N-ethylmaleimide 1 mM, no residual activity Flavobacterium frigidimaris
1.1.1.299 ZnCl2 1 mM, 27% residual activity Flavobacterium frigidimaris

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.37 0.0279
-
 NAD+ 10°C, pH 10.0 Flavobacterium frigidimaris
1.1.1.37 0.0286
-
 NAD+ 20°C, pH 10.0 Flavobacterium frigidimaris
1.1.1.37 0.0299
-
 NAD+ 30°C, pH 10.0 Flavobacterium frigidimaris
1.1.1.37 0.0382
-
 NAD+ 40°C, pH 10.0 Flavobacterium frigidimaris
1.1.1.37 0.269
-
 (S)-malate 20°C, pH 10.0 Flavobacterium frigidimaris
1.1.1.37 0.288
-
 (S)-malate 30°C, pH 10.0 Flavobacterium frigidimaris
1.1.1.37 0.543
-
 (S)-malate 10°C, pH 10.0 Flavobacterium frigidimaris
1.1.1.37 0.74
-
 (S)-malate 40°C, pH 10.0 Flavobacterium frigidimaris
1.1.1.299 0.028
-
 NAD+ 10°C Flavobacterium frigidimaris
1.1.1.299 0.029
-
 NAD+ 20°C Flavobacterium frigidimaris
1.1.1.299 0.03
-
 NAD+ 30°C Flavobacterium frigidimaris
1.1.1.299 0.038
-
 NAD+ 40°C Flavobacterium frigidimaris
1.1.1.299 0.269
-
 L-malate 20°C Flavobacterium frigidimaris
1.1.1.299 0.288
-
 L-malate 30°C Flavobacterium frigidimaris
1.1.1.299 0.543
-
 L-malate 10°C Flavobacterium frigidimaris
1.1.1.299 0.74
-
 L-malate 40°C Flavobacterium frigidimaris

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.1.1.37 32000
-
 4 * 32000, SDS-PAGE Flavobacterium frigidimaris
1.1.1.37 123000
-
 gel filtration Flavobacterium frigidimaris
1.1.1.299 32000
-
 4 * 32000, SDS-PAGE Flavobacterium frigidimaris
1.1.1.299 123000
-
 gel filtration Flavobacterium frigidimaris

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.37 Flavobacterium frigidimaris
-
-
-
1.1.1.37 Flavobacterium frigidimaris KUC-1
-
-
-
1.1.1.299 Flavobacterium frigidimaris
-
-
-
1.1.1.299 Flavobacterium frigidimaris KUC-1
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.37
-
 Flavobacterium frigidimaris

Reaction

EC Number Reaction Comment Organism Reaction ID
1.1.1.299 (S)-malate + NAD(P)+ = oxaloacetate + NAD(P)H + H+ ordered bi-bi mechanism Flavobacterium frigidimaris
a

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.1.1.37 176
-
-
 Flavobacterium frigidimaris
1.1.1.299 176
-
 30°C Flavobacterium frigidimaris

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.37 (S)-malate + NAD+ NAD+ binds first to the enzyme and then oxaloacetate is consequently released randomly via an ordered via an ordered bi-bi mechanism Flavobacterium frigidimaris oxaloacetate + NADH + H+
-
 r
1.1.1.37 (S)-malate + NAD+ NAD+ binds first to the enzyme and then oxaloacetate is consequently released randomly via an ordered via an ordered bi-bi mechanism Flavobacterium frigidimaris KUC-1 oxaloacetate + NADH + H+
-
 r
1.1.1.37 (S)-malate + NADP+
-
 Flavobacterium frigidimaris oxaloacetate + NADPH + H+
-
 ir
1.1.1.37 (S)-malate + NADP+
-
 Flavobacterium frigidimaris KUC-1 oxaloacetate + NADPH + H+
-
 ir
1.1.1.299 (S)-malate + NAD+
-
 Flavobacterium frigidimaris oxaloacetate + NADH + H+
-
 ?
1.1.1.299 (S)-malate + NAD+
-
 Flavobacterium frigidimaris KUC-1 oxaloacetate + NADH + H+
-
 ?
1.1.1.299 (S)-malate + NADP+
-
 Flavobacterium frigidimaris oxaloacetate + NADPH + H+
-
 ?
1.1.1.299 (S)-malate + NADP+
-
 Flavobacterium frigidimaris KUC-1 oxaloacetate + NADPH + H+
-
 ?
1.1.1.299 additional information enzyme shows pro-R stereospecificity for hydrogen transfer at the C4 position of the nicotinamide moiety of the coenzyme. No substrates are: D-malate, malonate, L-glutamate, L-aspartate, D,L-2-hydroxybutanoate, D,L-3-hydroxybutanoate, citrate, maleate, succinate, L-tartrate, L-threonine, L-serine, L-hydroxymalonate, D-glutamate, 2-oxocaproate, 2-oxoisovalerate, glyoxylate, 2-oxoglutarate, 2-oxobutanoate Flavobacterium frigidimaris ?
-
 ?
1.1.1.299 additional information enzyme shows pro-R stereospecificity for hydrogen transfer at the C4 position of the nicotinamide moiety of the coenzyme. No substrates are: D-malate, malonate, L-glutamate, L-aspartate, D,L-2-hydroxybutanoate, D,L-3-hydroxybutanoate, citrate, maleate, succinate, L-tartrate, L-threonine, L-serine, L-hydroxymalonate, D-glutamate, 2-oxocaproate, 2-oxoisovalerate, glyoxylate, 2-oxoglutarate, 2-oxobutanoate Flavobacterium frigidimaris KUC-1 ?
-
 ?
1.1.1.299 oxaloacetate + NADH + H+
-
 Flavobacterium frigidimaris L-malate + NAD+
-
 ?

Subunits

EC Number Subunits Comment Organism
1.1.1.37 tetramer 4 * 32000, SDS-PAGE Flavobacterium frigidimaris
1.1.1.299 tetramer 4 * 32000, SDS-PAGE Flavobacterium frigidimaris

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.1.37 40
-
-
 Flavobacterium frigidimaris
1.1.1.299 40
-
 highest initial velocity Flavobacterium frigidimaris

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
1.1.1.37 20 50 20°C: about 55% of maximal activity, 50°C: about 50% of maximal activity Flavobacterium frigidimaris
1.1.1.299 5 60
-
 Flavobacterium frigidimaris

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.1.1.37 35
-
 half-life: 26 min Flavobacterium frigidimaris
1.1.1.37 40
-
 half-life: 3 min Flavobacterium frigidimaris
1.1.1.37 45
-
 half-life: 0.6 min Flavobacterium frigidimaris
1.1.1.299 35
-
 half-life 26 min Flavobacterium frigidimaris
1.1.1.299 40
-
 half-life 3 min Flavobacterium frigidimaris

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1.1.37 1.73
-
 NAD+ 30°C, pH 10.0 Flavobacterium frigidimaris
1.1.1.37 41
-
 NAD+ 10°C, pH 10.0 Flavobacterium frigidimaris
1.1.1.37 41.3
-
 (S)-malate 10°C, pH 10.0 Flavobacterium frigidimaris
1.1.1.37 48.4
-
 (S)-malate 20°C, pH 10.0 Flavobacterium frigidimaris
1.1.1.37 48.6
-
 NAD+ 20°C, pH 10.0 Flavobacterium frigidimaris
1.1.1.37 83.2
-
 (S)-malate 30°C, pH 10.0 Flavobacterium frigidimaris
1.1.1.37 83.4
-
 NAD+ 30°C, pH 10.0 Flavobacterium frigidimaris
1.1.1.37 138
-
 (S)-malate 40°C, pH 10.0 Flavobacterium frigidimaris
1.1.1.37 139
-
 NAD+ 40°C, pH 10.0 Flavobacterium frigidimaris
1.1.1.299 41
-
 NAD+ 10°C Flavobacterium frigidimaris
1.1.1.299 41.3
-
 L-malate 10°C Flavobacterium frigidimaris
1.1.1.299 48.4
-
 L-malate 20°C Flavobacterium frigidimaris
1.1.1.299 48.6
-
 NAD+ 20°C Flavobacterium frigidimaris
1.1.1.299 83.2
-
 L-malate 30°C Flavobacterium frigidimaris
1.1.1.299 83.4
-
 NAD+ 30°C Flavobacterium frigidimaris
1.1.1.299 138
-
 L-malate 40°C Flavobacterium frigidimaris
1.1.1.299 139
-
 NAD+ 40°C Flavobacterium frigidimaris

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.1.37 8
-
 reduction of oxaloacetate Flavobacterium frigidimaris
1.1.1.37 10.5
-
 oxidation of malate Flavobacterium frigidimaris
1.1.1.299 8
-
 reduction of oxaloacetate Flavobacterium frigidimaris
1.1.1.299 10.5
-
 oxidation of malate Flavobacterium frigidimaris

pH Range

EC Number pH Minimum pH Maximum Comment Organism
1.1.1.299 4 10 reduction of oxaloacetate Flavobacterium frigidimaris
1.1.1.299 6.5 12 oxidation of malate Flavobacterium frigidimaris

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
1.1.1.37 4 10.5 stable Flavobacterium frigidimaris
1.1.1.299 4 10.5
-
 Flavobacterium frigidimaris

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.37 NAD+ the relative activity for NAD+ to that of NADP+ is about 43.9%. pro-R stereospecificity for hydrogen transfer at the C4 position of the nicotinamide moiety of the coenzyme Flavobacterium frigidimaris
1.1.1.37 NADH pro-R stereospecificity for hydrogen transfer at the C4 position of the nicotinamide moiety of the coenzyme Flavobacterium frigidimaris
1.1.1.37 NADP+ the relative activity for NAD+ to that of NADP+ is about 43.9%. No activity with NADPH Flavobacterium frigidimaris
1.1.1.299 additional information no cofactor: NADPH Flavobacterium frigidimaris
1.1.1.299 NAD+ relative activity of NAD+ to NADP+ is 43.9% Flavobacterium frigidimaris
1.1.1.299 NADH
-
 Flavobacterium frigidimaris
1.1.1.299 NADP+ relative activity of NAD+ to NADP+ is 43.9% Flavobacterium frigidimaris

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.1.1.299 289
-
 L-malate 30°C Flavobacterium frigidimaris
1.1.1.299 2790
-
 NAD+ 30°C Flavobacterium frigidimaris