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Literature summary for 1.5.1.3 extracted from

  • Redecke, L.; Brehm, M.A.; Bredehorst, R.
    Cloning and characterization of dihydrofolate reductase from a facultative alkaliphilic and halotolerant bacillus strain (2007), Extremophiles, 11, 75-83.
    View publication on PubMed
Search for more literature for 1.5.1.3

Cloned(Commentary)

Cloned (Comment) Organism
-
 Halalkalibacterium halodurans
expression in Escherichia coli Halalkalibacterium halodurans

Protein Variants

Protein Variants Comment Organism
additional information alkaliphilic properties of enzyme are diminished upon addition of a N-terminal His-tag Halalkalibacterium halodurans
additional information a C-terminal His-tag does not affect the catalytic properties of the enzyme, but results in loss of the thermal stability and of the alkali-stable phenotype Halalkalibacterium halodurans

General Stability

General Stability Organism
50% residual activity in 4 M urea Halalkalibacterium halodurans
stable in NaCl or KCl up to 0.3 M with slight activation below 0.1 M Halalkalibacterium halodurans

Inhibitors

Inhibitors Comment Organism Structure
methotrexate wild-type, 50% inhibition at 0.00031 mM Halalkalibacterium halodurans
additional information stable in up to 3 M urea; wild-type enzyme is not affected by presence of NaCl or KCl up to a concentration of 0.3 M, while the His-tagged enzyme is affected even at low concentrations Halalkalibacterium halodurans
trimethoprim wild-type, 50% inhibition at 0.0089 mM Halalkalibacterium halodurans

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0013
-
 7,8-dihydrofolate wild-type, pH 7.5, 37°C Halalkalibacterium halodurans
0.0029
-
 7,8-dihydrofolate His-tagged recombinant enzyme, pH 7.5, 37°C Halalkalibacterium halodurans
0.0029
-
 7,8-dihydrofolate His-tagged enzyme, pH 7.5, 37°C Halalkalibacterium halodurans
0.0059
-
 NADPH His-tagged recombinant enzyme, pH 7.5, 37°C Halalkalibacterium halodurans
0.0059
-
 NADPH His-tagged enzyme, pH 7.5, 37°C Halalkalibacterium halodurans
0.0082
-
 NADPH wild-type, pH 7.5, 37°C Halalkalibacterium halodurans

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
18855
-
 1 * 18855, mass spectrometry, dynamic light scattering Halalkalibacterium halodurans
20157
-
 x + 18855, wild-type, x * 20157, His-tagged enzyme, mass spectrometry Halalkalibacterium halodurans

Organism

Organism UniProt Comment Textmining
Halalkalibacterium halodurans
-
-
-
Halalkalibacterium halodurans
-
 expression in Escherichia coli
-
Halalkalibacterium halodurans C-125
-
-
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
122.9
-
 pH 7.5, 37°C Halalkalibacterium halodurans
122.9
-
 His-tagged enzyme, pH 7.5, 37°C Halalkalibacterium halodurans
138.4
-
 wild-type, pH 7.5, 37°C Halalkalibacterium halodurans

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
7,8-dihydrofolate + NADPH
-
 Halalkalibacterium halodurans 5,6,7,8-tetrahydrofolate + NADP+
-
 ?
7,8-dihydrofolate + NADPH
-
 Halalkalibacterium halodurans C-125 5,6,7,8-tetrahydrofolate + NADP+
-
 ?
7,8-dihydrofolate + NADPH + H+
-
 Halalkalibacterium halodurans 5,6,7,8-tetrahydrofolate + NADP+
-
 ?
7,8-dihydrofolate + NADPH + H+
-
 Halalkalibacterium halodurans C-125 5,6,7,8-tetrahydrofolate + NADP+
-
 ?

Subunits

Subunits Comment Organism
? x + 18855, wild-type, x * 20157, His-tagged enzyme, mass spectrometry Halalkalibacterium halodurans
monomer 1 * 18855, mass spectrometry, dynamic light scattering Halalkalibacterium halodurans

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
45
-
 His-tagged enzyme, stable up to Halalkalibacterium halodurans
45
-
 stable up to, His-tagged enzyme Halalkalibacterium halodurans
50
-
 wild-type, stable for 3 h Halalkalibacterium halodurans
50
-
 3 h, stable, wild-type Halalkalibacterium halodurans
52.5
-
 wild-type, halflife of 60 min, His-tagged recombinant enzyme, halflife 15 min Halalkalibacterium halodurans
52.5
-
 half-life 60 min for wild-type, 15 min for His-tagged enzyme, respectively Halalkalibacterium halodurans
60
-
 wild-type, complete inactivation above Halalkalibacterium halodurans

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6.75 7.75 His-tagged recombinant enzyme Halalkalibacterium halodurans
6.75 7.5 His-tagged enzyme Halalkalibacterium halodurans
7.25 8.75 wild-type Halalkalibacterium halodurans
7.25 8.25 broad, wild-type Halalkalibacterium halodurans

pH Range

pH Minimum pH Maximum Comment Organism
9
-
 rapid decrease in activity above, wild-type Halalkalibacterium halodurans

pH Stability

pH Stability pH Stability Maximum Comment Organism
9
-
 rapid decrease in stability above Halalkalibacterium halodurans

Cofactor

Cofactor Comment Organism Structure
NADP+
-
 Halalkalibacterium halodurans
NADPH
-
 Halalkalibacterium halodurans

IC50 Value

IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
0.00025
-
 His-tagged enzyme, pH 7.5, 37°C Halalkalibacterium halodurans methotrexate
0.00031
-
 wild-type, pH 7.5, 37°C Halalkalibacterium halodurans methotrexate
0.0089
-
 wild-type, pH 7.5, 37°C Halalkalibacterium halodurans trimethoprim
0.0101
-
 His-tagged enzyme, pH 7.5, 37°C Halalkalibacterium halodurans trimethoprim