Cloned (Comment) | Organism |
---|---|
functionally expressed in Escherichia coli as the N-terminal His6-tagged protein | Rauvolfia serpentina |
Protein Variants | Comment | Organism |
---|---|---|
D52A | inactive mutant enzyme | Rauvolfia serpentina |
H126A | inactive mutant enzyme | Rauvolfia serpentina |
K84A | inactive mutant enzyme | Rauvolfia serpentina |
additional information | site-directed mutagenesis of each of the functional residues of catalytic tetrad Asp52, Tyr57, Lys84, His126 to an alanine residue results in more than 97.8% loss of enzyme activity | Rauvolfia serpentina |
Y57A | inactive mutant enzyme | Rauvolfia serpentina |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.28 | - |
4-nitrobenzaldehyde | pH 8.5, 37°C | Rauvolfia serpentina | |
0.83 | - |
perakine | pH 8.5, 37°C | Rauvolfia serpentina |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
37159 | - |
x * 37159, calculated from sequence | Rauvolfia serpentina |
38000 | - |
x * 38000, SDS-PAGE | Rauvolfia serpentina |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
perakine + NADPH + H+ | Rauvolfia serpentina | the biosynthesis of raucaffrinoline from perakine is a side route of the ajmaline biosynthesis pathway (monoterpenoid indole alkaloid) | raucaffrinoline + NADP+ | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Rauvolfia serpentina | Q3L181 | - |
- |
Purification (Comment) | Organism |
---|---|
- |
Rauvolfia serpentina |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-nitrobenzaldehyde + NADPH + H+ | - |
Rauvolfia serpentina | 4-nitrobenzoate + NADP+ | - |
? | |
additional information | the enzyme also catalyses the reduction of coniferyl aldehyde, sinapyl aldehyde, 4-coumaryl aldehyde, 3-(3,4,5-trimethoxyphenyl)propenal | Rauvolfia serpentina | ? | - |
? | |
perakine + NADPH + H+ | the biosynthesis of raucaffrinoline from perakine is a side route of the ajmaline biosynthesis pathway (monoterpenoid indole alkaloid) | Rauvolfia serpentina | raucaffrinoline + NADP+ | - |
? | |
perakine + NADPH + H+ | NADPH can not be replaced by NADH. In presence of the oxidized cofactor NADP+ the reverse reaction of the reductase is low, with less than 0.5% relative activity of the forward reaction | Rauvolfia serpentina | raucaffrinoline + NADP+ | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 38000, SDS-PAGE | Rauvolfia serpentina |
? | x * 37159, calculated from sequence | Rauvolfia serpentina |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Rauvolfia serpentina |
50 | - |
- |
Rauvolfia serpentina |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.014 | - |
perakine | pH 8.5, 37°C | Rauvolfia serpentina | |
8.26 | - |
4-nitrobenzaldehyde | pH 8.5, 37°C | Rauvolfia serpentina |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
- |
Rauvolfia serpentina |
8.5 | - |
assay at | Rauvolfia serpentina |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
6.5 | 8.5 | half-maximal activity | Rauvolfia serpentina |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADPH | can not be replaced by NADH | Rauvolfia serpentina |
Organism | Comment | pI Value Maximum | pI Value |
---|---|---|---|
Rauvolfia serpentina | calculated from sequence | - |
5.9 |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.016 | - |
perakine | pH 8.5, 37°C | Rauvolfia serpentina | |
29.5 | - |
4-nitrobenzaldehyde | pH 8.5, 37°C | Rauvolfia serpentina |