Cloned (Comment) | Organism |
---|---|
salR, DNA and amino acid sequence determination and analysis, overexpression of the His-tagged enzyme in Escherichia coli | Papaver bracteatum |
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 | 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 | Comment | Organism | Structure |
---|---|---|---|
salutaridine | - |
Papaver bracteatum |
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 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 | 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 | UniProt | Comment | Textmining |
---|---|---|---|
Papaver bracteatum | A4UHT7 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged enzyme from Escherichia coli by cobalt affinity chromatography | Papaver bracteatum |
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 | 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 | 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 Maximum [°C] | Comment | Organism |
---|---|---|---|
40 | - |
broad optimum | Papaver bracteatum |
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 Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6 | - |
reduction reaction | Papaver bracteatum |
9.5 | - |
oxidation reaction | Papaver bracteatum |
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 | 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 maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.14 | - |
salutaridine | pH 6.0, recombinant enzyme | Papaver bracteatum |
Organism | Comment | pI Value Maximum | pI Value |
---|---|---|---|
Papaver bracteatum | sequence calculation | - |
4.72 |