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Literature summary 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)

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

Protein Variants

EC Number Protein Variants Comment Organism
1.1.1.317 D52A inactive mutant enzyme Rauvolfia serpentina
1.1.1.317 H126A inactive mutant enzyme Rauvolfia serpentina
1.1.1.317 K84A inactive mutant enzyme Rauvolfia serpentina
1.1.1.317 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
1.1.1.317 Y57A inactive mutant enzyme Rauvolfia serpentina

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.317 0.28
-
 4-nitrobenzaldehyde pH 8.5, 37°C Rauvolfia serpentina
1.1.1.317 0.83
-
 perakine pH 8.5, 37°C Rauvolfia serpentina

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.1.1.317 37159
-
 x * 37159, calculated from sequence Rauvolfia serpentina
1.1.1.317 38000
-
 x * 38000, SDS-PAGE Rauvolfia serpentina

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.1.1.317 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

EC Number Organism UniProt Comment Textmining
1.1.1.317 Rauvolfia serpentina Q3L181
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.317
-
 Rauvolfia serpentina

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.317 4-nitrobenzaldehyde + NADPH + H+
-
 Rauvolfia serpentina 4-nitrobenzoate + NADP+
-
 ?
1.1.1.317 additional information the enzyme also catalyses the reduction of coniferyl aldehyde, sinapyl aldehyde, 4-coumaryl aldehyde, 3-(3,4,5-trimethoxyphenyl)propenal Rauvolfia serpentina ?
-
 ?
1.1.1.317 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+
-
 ?
1.1.1.317 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

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

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.1.317 37
-
 assay at Rauvolfia serpentina
1.1.1.317 50
-
-
 Rauvolfia serpentina

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1.1.317 0.014
-
 perakine pH 8.5, 37°C Rauvolfia serpentina
1.1.1.317 8.26
-
 4-nitrobenzaldehyde pH 8.5, 37°C Rauvolfia serpentina

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.1.317 7
-
-
 Rauvolfia serpentina
1.1.1.317 8.5
-
 assay at Rauvolfia serpentina

pH Range

EC Number pH Minimum pH Maximum Comment Organism
1.1.1.317 6.5 8.5 half-maximal activity Rauvolfia serpentina

Cofactor

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

pI Value

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

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.1.1.317 0.016
-
 perakine pH 8.5, 37°C Rauvolfia serpentina
1.1.1.317 29.5
-
 4-nitrobenzaldehyde pH 8.5, 37°C Rauvolfia serpentina