Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 1.1.1.248 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)

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

Crystallization (Commentary)

Crystallization (Comment) Organism
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

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

Inhibitors

Inhibitors Comment Organism Structure
salutaridine
-
Papaver bracteatum

KM Value [mM]

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

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
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
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)

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

Organism

Organism UniProt Comment Textmining
Papaver bracteatum A4UHT7
-
-

Purification (Commentary)

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

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information the enzyme is involved in the biosynthesis of morphine Papaver bracteatum ?
-
?
additional information interaction of benzylisoquinoline alkaloids with enzymes, modeling, overview Papaver bracteatum ?
-
?
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
salutaridine + NADH + H+
-
Papaver bracteatum salutaridinol + NAD+
-
r
salutaridine + NADPH + H+
-
Papaver bracteatum (7S)-salutaridinol + NADP+
-
r

Subunits

Subunits Comment Organism
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

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

Temperature Optimum [°C]

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

Turnover Number [1/s]

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

pH Optimum

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

pH Range

pH Minimum pH Maximum Comment Organism
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

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

Ki Value [mM]

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

pI Value

Organism Comment pI Value Maximum pI Value
Papaver bracteatum sequence calculation
-
4.72