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Literature summary extracted from

  • Geissler, R.; Brandt, W.; Ziegler, J.
    Molecular modeling and site-directed mutagenesis reveal the benzylisoquinoline binding site of the short-chain dehydrogenase/reductase salutaridine reductase (2007), Plant Physiol., 143, 1493-1503.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.248 salR, DNA and amino acid sequence determination and analysis, overexpression of the His-tagged enzyme in Escherichia coli Papaver bracteatum

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
1.1.1.248 construction of a homology model of the Papaver bracteatum SalR based on the X-ray structure of human carbonyl reductase 1, overview Papaver bracteatum

Protein Variants

EC Number Protein Variants Comment Organism
1.1.1.248 F104A site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Papaver bracteatum
1.1.1.248 K240E site-directed mutagenesis, inactive mutant Papaver bracteatum
1.1.1.248 L266A site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Papaver bracteatum
1.1.1.248 L266S site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Papaver bracteatum
1.1.1.248 L266V site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Papaver bracteatum
1.1.1.248 M271T site-directed mutagenesis, inactive mutant Papaver bracteatum
1.1.1.248 N152A site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Papaver bracteatum
1.1.1.248 N272T site-directed mutagenesis, inactive mutant Papaver bracteatum
1.1.1.248 R44E site-directed mutagenesis, the mutant enzyme shows altered cofactor specificity and utilizes also NADH in contrast to the wild-type enzyme Papaver bracteatum
1.1.1.248 R48E site-directed mutagenesis, the mutant enzyme shows altered cofactor specificity and utilizes also NADH in contrast to the wild-type enzyme Papaver bracteatum
1.1.1.248 S180A site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme Papaver bracteatum
1.1.1.248 V106A site-directed mutagenesis, the mutant shows about 2fold increased activity compared to the wild-type enzyme Papaver bracteatum
1.1.1.248 Y236F site-directed mutagenesis, inactive mutant Papaver bracteatum

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.1.1.248 salutaridine
-
 Papaver bracteatum

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.248 additional information
-
 additional information kinetics of wild-type and mutant enzymes, overview Papaver bracteatum
1.1.1.248 0.0015
-
 (7S)-salutaridinol pH 6.0, 40°C, recombinant enzyme Papaver bracteatum
1.1.1.248 0.0035
-
 NADPH pH 6.0, 40°C, recombinant enzyme, with salutaridine Papaver bracteatum
1.1.1.248 0.007
-
 NADP+ pH 6.0, 40°C, recombinant enzyme, with salutaridinol Papaver bracteatum
1.1.1.248 0.0079
-
 salutaridine pH 6.0, 40°C, recombinant enzyme Papaver bracteatum
1.1.1.248 1.19
-
 NADH pH 6.0, 40°C, recombinant enzyme, with salutaridine Papaver bracteatum

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.1.1.248 34050
-
 1 * 36600, recombinant His-tagged enzyme, SDS-PAGE, 1 * 34050, sequence calculation, the tertiary structure comprises the typical short-chain dehydrogenase/reductase family alpha/beta folding pattern including the four additional helices alphaF-1 to alphaF-4 assumed to prevent the dimerization, modeling and analysis, overview Papaver bracteatum
1.1.1.248 36600
-
 1 * 36600, recombinant His-tagged enzyme, SDS-PAGE, 1 * 34050, sequence calculation, the tertiary structure comprises the typical short-chain dehydrogenase/reductase family alpha/beta folding pattern including the four additional helices alphaF-1 to alphaF-4 assumed to prevent the dimerization, modeling and analysis, overview Papaver bracteatum

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.1.1.248 additional information Papaver bracteatum the enzyme is involved in the biosynthesis of morphine ?
-
 ?
1.1.1.248 salutaridine + NADPH + H+ Papaver bracteatum
-
 (7S)-salutaridinol + NADP+
-
 r

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.248 Papaver bracteatum A4UHT7
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.248 recombinant His-tagged enzyme from Escherichia coli by cobalt affinity chromatography Papaver bracteatum

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.248 additional information the enzyme is involved in the biosynthesis of morphine Papaver bracteatum ?
-
 ?
1.1.1.248 additional information interaction of benzylisoquinoline alkaloids with enzymes, modeling, overview Papaver bracteatum ?
-
 ?
1.1.1.248 salutaridine + NAD(P)H + H+ NADPH is the highly preferred cofactor, the enzyme shows high substrate specificity, no activity with tropinone, (2)-menthone, codeinone and dehydroreticulinium ion, or nordehydroreticuline. Asp152, Ser180, Tyr236, and Lys240 are involved in the proton transfer system for the reduction of salutaridine, substrate-active site binding structure, overview Papaver bracteatum (7S)-salutaridinol + NAD(P)+ the enzyme produces only the (7S)-salutaridinol stereoisomer as product, not the stereoisomer 7-epi-salutaridinol r
1.1.1.248 salutaridine + NADH + H+
-
 Papaver bracteatum salutaridinol + NAD+
-
 r
1.1.1.248 salutaridine + NADPH + H+
-
 Papaver bracteatum (7S)-salutaridinol + NADP+
-
 r

Subunits

EC Number Subunits Comment Organism
1.1.1.248 monomer 1 * 36600, recombinant His-tagged enzyme, SDS-PAGE, 1 * 34050, sequence calculation, the tertiary structure comprises the typical short-chain dehydrogenase/reductase family alpha/beta folding pattern including the four additional helices alphaF'-1 to alphaF'-4 assumed to prevent the dimerization, modeling and analysis, overview Papaver bracteatum

Synonyms

EC Number Synonyms Comment Organism
1.1.1.248 More the enzyme belongs to the NADPH-dependent short-chain dehydrogenase/reductase family Papaver bracteatum
1.1.1.248 SalR
-
 Papaver bracteatum

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.1.248 40
-
 broad optimum Papaver bracteatum

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1.1.248 2.1
-
 NADPH pH 6.0, 40°C, recombinant enzyme, with salutaridine Papaver bracteatum
1.1.1.248 2.3
-
 salutaridine pH 6.0, 40°C, recombinant enzyme Papaver bracteatum
1.1.1.248 3.7
-
 NADH pH 6.0, 40°C, recombinant enzyme, with salutaridine Papaver bracteatum
1.1.1.248 21.6
-
 (7S)-salutaridinol pH 6.0, 40°C, recombinant enzyme Papaver bracteatum
1.1.1.248 21.9
-
 NADP+ pH 6.0, 40°C, recombinant enzyme, with salutaridinol Papaver bracteatum

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.1.248 6
-
 reduction reaction Papaver bracteatum
1.1.1.248 9.5
-
 oxidation reaction Papaver bracteatum

pH Range

EC Number pH Minimum pH Maximum Comment Organism
1.1.1.248 4.5 7.5 the enzyme shows a steep decrease by 90% of activity within 1.5 pH units on both sides of the optimum at pH 6.0 for the reduction reaction Papaver bracteatum

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.248 NAD+
-
 Papaver bracteatum
1.1.1.248 NADH strong preference for NADPH over NADH Papaver bracteatum
1.1.1.248 NADP+
-
 Papaver bracteatum
1.1.1.248 NADPH strong preference for NADPH over NADH Papaver bracteatum

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
1.1.1.248 0.14
-
 salutaridine pH 6.0, recombinant enzyme Papaver bracteatum

pI Value

EC Number Organism Comment pI Value Maximum pI Value
1.1.1.248 Papaver bracteatum sequence calculation
-
 4.72