Cloned (Comment) | Organism |
---|---|
a variant (Xet-3) of dihydrofolate reductase from Escherichia coli constructed by introducing residues located at the Thermotoga maritima dihydrofolate reductase (TmDHFR) dimer interface is expressed in Escherichia coli BL21(DE3) cells | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
additional information | preparation of a dimeric variant (Xet-3) of dihydrofolate reductase from Escherichia coli by introducing residues located at the Thermotoga maritima dihydrofolate reductase (TmDHFR) dimer interface increases the melting temperature to about 60°C, approximately 9°C higher than that measured for the Escherichia coli enzyme. The steady-state and pre-steady-state rate constants measured for Xet-3 were similar to those of dimeric TmDHFR but significantly lower than those of the parent dihydrofolate reductase from Escherichia coli | Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0007 | - |
7,8-dihydrofolate | pH 7, 20°C, wild-type enzyme | Escherichia coli | |
0.0027 | - |
7,8-dihydrofolate | pH 7, 20°C, monomeric variant (Xet-3) of dihydrofolate reductase from Escherichia coli prepared by introducing residues located at the Thermotoga maritima dihydrofolate reductase dimer interface | Escherichia coli | |
0.0048 | - |
NADPH | pH 7, 20°C, wild-type enzyme | Escherichia coli | |
0.0077 | - |
7,8-dihydrofolate | pH 7, 20°C, dimeric variant (Xet-3) of dihydrofolate reductase from Escherichia coli prepared by introducing residues located at the Thermotoga maritima dihydrofolate reductase dimer interface | Escherichia coli | |
0.0195 | - |
NADPH | pH 7, 20°C, monomeric variant (Xet-3) of dihydrofolate reductase from Escherichia coli prepared by introducing residues located at the Thermotoga maritima dihydrofolate reductase dimer interface | Escherichia coli | |
0.0199 | - |
NADPH | pH 7, 20°C, dimeric variant (Xet-3) of dihydrofolate reductase from Escherichia coli prepared by introducing residues located at the Thermotoga maritima dihydrofolate reductase dimer interface | Escherichia coli |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | - |
- |
- |
Thermotoga maritima | Q60034 | - |
- |
Thermotoga maritima DSM 3109 | Q60034 | - |
- |
Purification (Comment) | Organism |
---|---|
- |
Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
7,8-dihydrofolate + NADPH + H+ | - |
Escherichia coli | 5,6,7,8-tetrahydrofolate + NADP+ | - |
? | |
7,8-dihydrofolate + NADPH + H+ | - |
Thermotoga maritima | 5,6,7,8-tetrahydrofolate + NADP+ | - |
? | |
7,8-dihydrofolate + NADPH + H+ | - |
Thermotoga maritima DSM 3109 | 5,6,7,8-tetrahydrofolate + NADP+ | - |
? |
Subunits | Comment | Organism |
---|---|---|
homodimer | - |
Thermotoga maritima |
monomer | preparation of a dimeric variant (Xet-3) of dihydrofolate reductase from Escherichia coli by introducing residues located at the Thermotoga maritima dihydrofolate reductase dimer interface | Escherichia coli |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
60 | - |
preparation of a dimeric variant (Xet-3) of dihydrofolate reductase from Escherichia coli by introducing residues located at the Thermotoga maritima dihydrofolate reductase dimer interface increases the melting temperature to about 60°C, approximately 9°C higher than that measured for the Escherichia coli enzyme | Escherichia coli |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.003 | - |
7,8-dihydrofolate | pH 7, 20°C, mutant enzyme V11D | Thermotoga maritima | |
0.05 | - |
NADPH | pH 7, 20°C, dimeric variant (Xet-3) of dihydrofolate reductase from Escherichia coli prepared by introducing residues located at the Thermotoga maritima dihydrofolate reductase dimer interface | Escherichia coli | |
0.05 | - |
7,8-dihydrofolate | pH 7, 20°C, dimeric variant (Xet-3) of dihydrofolate reductase from Escherichia coli prepared by introducing residues located at the Thermotoga maritima dihydrofolate reductase dimer interface | Escherichia coli | |
0.06 | - |
7,8-dihydrofolate | pH 7, 20°C, wild-type enzyme | Thermotoga maritima | |
1.39 | - |
NADPH | pH 7, 20°C, monomeric variant (Xet-3) of dihydrofolate reductase from Escherichia coli prepared by introducing residues located at the Thermotoga maritima dihydrofolate reductase dimer interface | Escherichia coli | |
1.39 | - |
7,8-dihydrofolate | pH 7, 20°C, monomeric variant (Xet-3) of dihydrofolate reductase from Escherichia coli prepared by introducing residues located at the Thermotoga maritima dihydrofolate reductase dimer interface | Escherichia coli | |
12.6 | - |
NADPH | pH 7, 20°C, wild-type enzyme | Escherichia coli | |
12.6 | - |
7,8-dihydrofolate | pH 7, 20°C, wild-type enzyme | Escherichia coli |