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Literature summary for 1.5.1.3 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

Activating Compound Comment Organism Structure
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)

Cloned (Comment) Organism
-
Geobacillus stearothermophilus

Inhibitors

Inhibitors Comment Organism Structure
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]

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

Molecular Weight [Da]

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

Organism

Organism UniProt Comment Textmining
Geobacillus stearothermophilus
-
-
-

Reaction

Reaction Comment Organism Reaction ID
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

Substrates and Products (Substrate)

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

Subunits

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

Temperature Optimum [°C]

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

Temperature Stability [°C]

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

Turnover Number [1/s]

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

Cofactor

Cofactor Comment Organism Structure
NADP+
-
Geobacillus stearothermophilus
NADPH
-
Geobacillus stearothermophilus

pI Value

Organism Comment pI Value Maximum pI Value
Geobacillus stearothermophilus isoelectric focusing
-
8.2