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Literature summary extracted from

  • Kim, H.S.; Damo, S.M.; Lee, S.Y.; Wemmer, D.; Klinman, J.P.
    Structure and hydride transfer mechanism of a moderate thermophilic dihydrofolate reductase from Bacillus stearothermophilus and comparison to its mesophilic and hyperthermophilic homologues (2005), Biochemistry, 44, 11428-11439.
    View publication on PubMed

Activating Compound

EC Number Activating Compound Comment Organism Structure
1.5.1.3 NaCl up to 20% increase in activity up to 0.2 M, activity declines above and falls to zero at 0.8 M Geobacillus stearothermophilus

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.5.1.3
-
 Geobacillus stearothermophilus

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.5.1.3 NaCl up to 20% increase in activity up to 0.2 M, activity declines above and falls to zero at 0.8 M Geobacillus stearothermophilus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.5.1.3 0.0037
-
 7,8-dihydrofolate pH 7.0, 60°C Geobacillus stearothermophilus
1.5.1.3 0.0104
-
 NADPH pH 7.0, 60°C Geobacillus stearothermophilus

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.5.1.3 15000
-
 gel filtration Geobacillus stearothermophilus
1.5.1.3 18694
-
 1 * 18694, electrospray mass spectrometry and calculated, 1 * 20000, SDS-PAGE Geobacillus stearothermophilus
1.5.1.3 20000
-
 1 * 18694, electrospray mass spectrometry and calculated, 1 * 20000, SDS-PAGE Geobacillus stearothermophilus

Organism

EC Number Organism UniProt Comment Textmining
1.5.1.3 Geobacillus stearothermophilus
-
-
-

Reaction

EC Number Reaction Comment Organism Reaction ID
1.5.1.3 5,6,7,8-tetrahydrofolate + NADP+ = 7,8-dihydrofolate + NADPH + H+ ordered kinetic mechanism at elevated temperature around 60°C, with NADPH binding first. More random mechanism at temperatures around 20°C with greatly reduced Km-value for 7,8-dihydrofolate. Rate-limiting hydride-transfer with a moderate enthalpy of activiation Geobacillus stearothermophilus
a

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.5.1.3 7,8-dihydrofolate + NADPH
-
 Geobacillus stearothermophilus 5,6,7,8-tetrahydrofolate + NADP+
-
 ?

Subunits

EC Number Subunits Comment Organism
1.5.1.3 monomer 1 * 18694, electrospray mass spectrometry and calculated, 1 * 20000, SDS-PAGE Geobacillus stearothermophilus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.5.1.3 65
-
 at pH 9 Geobacillus stearothermophilus
1.5.1.3 75
-
 at pH 7 Geobacillus stearothermophilus

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.5.1.3 60
-
 in presence of 5fold excess of NADPH or substrate, 70-80% residual activity after 1 h Geobacillus stearothermophilus
1.5.1.3 64
-
 rapid loss of activity in absence of substrate above 64°C Geobacillus stearothermophilus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.5.1.3 218
-
 NADPH pH 7.0, 60°C Geobacillus stearothermophilus

Cofactor

EC Number Cofactor Comment Organism Structure
1.5.1.3 NADP+
-
 Geobacillus stearothermophilus
1.5.1.3 NADPH
-
 Geobacillus stearothermophilus

pI Value

EC Number Organism Comment pI Value Maximum pI Value
1.5.1.3 Geobacillus stearothermophilus isoelectric focusing
-
 8.2