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Literature summary for 1.1.1.317 extracted from

  • Sun, L.; Ruppert, M.; Sheludko, Y.; Warzecha, H.; Zhao, Y.; Stöckigt, J.
    Purification, cloning, functional expression and characterization of perakine reductase: the first example from the AKR enzyme family, extending the alkaloidal network of the plant Rauvolfia (2008), Plant Mol. Biol., 67, 455-467.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
functionally expressed in Escherichia coli as the N-terminal His6-tagged protein Rauvolfia serpentina

Protein Variants

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 [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 [Da] Molecular Weight Maximum [Da] Comment Organism
37159
-
x * 37159, calculated from sequence Rauvolfia serpentina
38000
-
x * 38000, SDS-PAGE Rauvolfia serpentina

Natural Substrates/ Products (Substrates)

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

Organism UniProt Comment Textmining
Rauvolfia serpentina Q3L181
-
-

Purification (Commentary)

Purification (Comment) Organism
-
Rauvolfia serpentina

Substrates and Products (Substrate)

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

Subunits Comment Organism
? x * 38000, SDS-PAGE Rauvolfia serpentina
? x * 37159, calculated from sequence Rauvolfia serpentina

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
assay at Rauvolfia serpentina
50
-
-
Rauvolfia serpentina

Turnover Number [1/s]

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

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
-
Rauvolfia serpentina
8.5
-
assay at Rauvolfia serpentina

pH Range

pH Minimum pH Maximum Comment Organism
6.5 8.5 half-maximal activity Rauvolfia serpentina

Cofactor

Cofactor Comment Organism Structure
NADPH can not be replaced by NADH Rauvolfia serpentina

pI Value

Organism Comment pI Value Maximum pI Value
Rauvolfia serpentina calculated from sequence
-
5.9

kcat/KM [mM/s]

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