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Literature summary for 1.5.1.3 extracted from
- High expression in Escherichia coli of the gene coding for dihydrofolate reductase of the extremely halophilic archaebacterium Haloferax volcanii. Reconstitution of the active enzyme and mutation studies (1993), Eur. J. Biochem., 216, 199-203.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| overexpression of wild-type and mutant enzymes in Escherichia coli | Haloferax volcanii |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| A31K | site-directed mutagenesis | Haloferax volcanii |
| L30K | site-directed mutagenesis | Haloferax volcanii |
| L30K/A31K | site-directed mutagenesis | Haloferax volcanii |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| additional information | - |
additional information | overview Km-values of wild-type and mutant enzymes decreasing with increasing concentration of KCl | Haloferax volcanii | |
| 0.08 | - |
7,8-dihydrofolate | lowest Km value in 3.0 M KCl, recombinant wild-type enzyme | Haloferax volcanii |  |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Haloferax volcanii | - |
extremely halophilic archaebacterium | - |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| large quantities of recombinant wild-type enzyme from Escherichia coli, purification of mutants L30K, A31K and double mutant L30K/A31K recombinant from Escherichia coli | Haloferax volcanii |
Renatured (Commentary)
| Renatured (Comment) | Organism |
|---|---|
| very fast and total reconstitution of active recombinant wild-type enzyme from inclusion bodies due to overexpression in E. coli by 6 M guanidine hydrochloride followed by dilution into 1 M NaCl or KCl solution | Haloferax volcanii |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 7,8-dihydrofolate + NADPH | - |
Haloferax volcanii | 5,6,7,8-tetrahydrofolate + NADP+ | - |
? | |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 3.14 | - |
7,8-dihydrofolate | recombinant wild-type enzyme | Haloferax volcanii | |
| 4.97 | - |
7,8-dihydrofolate | recombinant mutant A31K | Haloferax volcanii | |
| 9.5 | - |
7,8-dihydrofolate | recombinant double mutant L30K/A31K | Haloferax volcanii | |
| 100 | - |
7,8-dihydrofolate | recombinant mutant L30K | Haloferax volcanii | |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| NADPH | - |
Haloferax volcanii | |
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