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Literature summary extracted from

  • Blecher, O.; Goldman, S.; Mevarech, M.
    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.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.5.1.3 overexpression of wild-type and mutant enzymes in Escherichia coli Haloferax volcanii

Protein Variants

EC Number Protein Variants Comment Organism
1.5.1.3 A31K site-directed mutagenesis Haloferax volcanii
1.5.1.3 L30K site-directed mutagenesis Haloferax volcanii
1.5.1.3 L30K/A31K site-directed mutagenesis Haloferax volcanii

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.5.1.3 additional information
-
 additional information overview Km-values of wild-type and mutant enzymes decreasing with increasing concentration of KCl Haloferax volcanii
1.5.1.3 0.08
-
 7,8-dihydrofolate lowest Km value in 3.0 M KCl, recombinant wild-type enzyme Haloferax volcanii

Organism

EC Number Organism UniProt Comment Textmining
1.5.1.3 Haloferax volcanii
-
 extremely halophilic archaebacterium
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.5.1.3 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)

EC Number Renatured (Comment) Organism
1.5.1.3 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)

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

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.5.1.3 3.14
-
 7,8-dihydrofolate recombinant wild-type enzyme Haloferax volcanii
1.5.1.3 4.97
-
 7,8-dihydrofolate recombinant mutant A31K Haloferax volcanii
1.5.1.3 9.5
-
 7,8-dihydrofolate recombinant double mutant L30K/A31K Haloferax volcanii
1.5.1.3 100
-
 7,8-dihydrofolate recombinant mutant L30K Haloferax volcanii

Cofactor

EC Number Cofactor Comment Organism Structure
1.5.1.3 NADPH
-
 Haloferax volcanii