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Literature summary extracted from

  • Supangat, S.; Choi, Y.K.; Park, Y.S.; Son, D.; Han, C.D.; Lee, K.H.
    Expression, purification, crystallization and preliminary X-ray analysis of sepiapterin reductase from Chlorobium tepidum (2005), Acta Crystallogr. Sect. F, 61, 202-204.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.153 expression of His-tagged native and selenomethionine-labeled enzyme in Escherichia coli strain BL21(DE3) Chlorobaculum tepidum
1.1.1.325 expression of His-tagged wild-type and selenomethionine-substituted enzymes in Escherichia coli strain BL21(DE3) Chlorobaculum tepidum

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
1.1.1.153 purified native and selenomethionine-labeled enzyme, hanging drop vapour diffusion microbatch method, 18°C, 0.004 ml protein solution containing 10 mg/ml protein in 20 mM Tris-HCl, pH 8.0, is mixed with 0.001 ml of a mixture of 0.004 ml or reservoir solution containing 0.2 M MgCl2, 0.1 M Tris-HCl, pH 8.5, and 34% PEG 400, and 0.001 ml additive solution 1 M guanidinium hydrochloride, 2 days, X-ray diffraction structure determination and analysis at 2.1 A resolution Chlorobaculum tepidum
1.1.1.325 purified recombinant His-tagged wild-type and selenomethionine-substituted enzymes, hanging drop vapour diffusion method at 18°C, mixing of 0.004 ml of protein solution containing 10 mg/ml in 20 mM Tris-HCl, pH 8.0, by ultrafiltration, with 0.001 ml of reservoir solution containing 0.2 M MgCl2, 0.1 M Tris-HCl, pH 8.5, 34% PEG 400, and 0.2 M guanidine hydrochloride, screening and method optimization, crystals of the wild-type enzyme are soaked in reservoir solution containing 1 mM NADP+ and 1 mM sepiapterin or containing 1 mM NADP+ and 1 mM N-acetylserotonin, respectively, X-ray diffraction structure determination and analysis at 2.1 A resolution Chlorobaculum tepidum

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.1.1.153 6-pyruvoyl tetrahydropterin + 2 NADPH + 2 H+ Chlorobaculum tepidum last step of the de novo tetrahydrobiopterin biosynthesis initiating from GTP L-threo-(6R,1'S,2'S)-5,6,7,8-tetrahydrobiopterin + NADP+
-
 ?
1.1.1.325 6-pyruvoyl-5,6,7,8-tetrahydropterin + 3 NADPH + 3 H+ Chlorobaculum tepidum
-
 L-threo-7,8-dihydrobiopterin + 3 NADP+
-
 ?

Organism

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

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.153 recombinant His-tagged native and selenomethionine-labeled enzyme from Escherichia coli strain BL21(DE3) by nickel affinity and anion exchange chromatography, followed by gel filtration Chlorobaculum tepidum
1.1.1.325 recombinant His-tagged wild-type and selenomethionine-substituted enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, ultrafiltration, anion exchange chromatography, and gel filtration Chlorobaculum tepidum

Reaction

EC Number Reaction Comment Organism Reaction ID
1.1.1.153 L-erythro-7,8-dihydrobiopterin + NADP+ = sepiapterin + NADPH + H+ active site structure Chlorobaculum tepidum
a

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+ last step of the de novo tetrahydrobiopterin biosynthesis initiating from GTP Chlorobaculum tepidum L-threo-(6R,1'S,2'S)-5,6,7,8-tetrahydrobiopterin + NADP+
-
 ?
1.1.1.153 6-pyruvoyl tetrahydropterin + 2 NADPH + 2 H+ two reductive steps oxidizing NADPH Chlorobaculum tepidum L-threo-(6R,1'S,2'S)-5,6,7,8-tetrahydrobiopterin + NADP+
-
 ?
1.1.1.325 6-pyruvoyl-5,6,7,8-tetrahydropterin + 3 NADPH + 3 H+
-
 Chlorobaculum tepidum L-threo-7,8-dihydrobiopterin + 3 NADP+
-
 ?

Synonyms

EC Number Synonyms Comment Organism
1.1.1.153 SR
-
 Chlorobaculum tepidum
1.1.1.325 sepiapterin reductase
-
 Chlorobaculum tepidum

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.153 NADPH
-
 Chlorobaculum tepidum
1.1.1.325 NADPH dependent on Chlorobaculum tepidum