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Literature summary extracted from

  • Supangat, S.; Park, S.O.; Seo, K.H.; Lee, S.Y.; Park, Y.S.; Lee, K.H.
    Role of Phe-99 and Trp-196 of sepiapterin reductase from Chlorobium tepidum in the production of L-threo-tetrahydrobiopterin (2008), Acta Biochim. Biophys. Sin. (Shanghai), 40, 513-518.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.153 expressed in Escherichia coli BL21(DE3) cells Chlorobaculum tepidum

Protein Variants

EC Number Protein Variants Comment Organism
1.1.1.153 F99A the mutant shows 3.9fold higher Km and lower Vmax (8.95%) than the wild type enzyme Chlorobaculum tepidum
1.1.1.153 W196A the mutant shows 8.7fold higher Km and lower Vmax (5.64%) than the wild type enzyme Chlorobaculum tepidum

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.153 0.00018
-
 sepiapterin wild type enzyme Chlorobaculum tepidum
1.1.1.153 0.00072
-
 sepiapterin mutant enzyme F99A Chlorobaculum tepidum
1.1.1.153 0.0016
-
 sepiapterin mutant enzyme W196A Chlorobaculum tepidum

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.153 Chlorobaculum tepidum
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.153 Ni-NTA bead chromatography, MonoQ column chromatography, and Superdex 200 gel filtration Chlorobaculum tepidum

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.153 6-pyruvoyl tetrahydropterin + 2 NADPH + 2 H+
-
 Chlorobaculum tepidum L-threo-(6R,1'S,2'S)-5,6,7,8-tetrahydrobiopterin + NADP+
-
 ?
1.1.1.153 sepiapterin + NADPH + H+
-
 Chlorobaculum tepidum 7,8-dihydrobiopterin + NADP+
-
 ?

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.1.1.153 72
-
 the melting temperature of the wild type enzyme is at 72.77°C Chlorobaculum tepidum

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.153 NADPH
-
 Chlorobaculum tepidum