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Literature summary for 3.2.1.125 extracted from

  • Xia, L.; Ruppert, M.; Wang, M.; Panjikar, S.; Lin, H.; Rajendran, C.; Barleben, L.; Stoeckigt, J.
    Structures of alkaloid biosynthetic glucosidases decode substrate specificity (2012), ACS Chem. Biol., 7, 226-234.
    View publication on PubMed

Crystallization (Commentary)

Crystallization (Comment) Organism
wild type and mutant enzyme E186Q in complex with the substrate 1,2-(S)-dihydroraucaffricine and secologanin, hanging drop vapor diffusion method, using 0.01-0.3 M ammonium sulfate, 0.1 M sodium acetate, pH 4.5-5.0, 9-12% (w/v) PEG 4000 as precipitant buffer, and 20°C Rauvolfia serpentina

Protein Variants

Protein Variants Comment Organism
E186D inactive Rauvolfia serpentina
E186Q inactive Rauvolfia serpentina
E186Q/E420Q inactive Rauvolfia serpentina
E420Q the mutant shows 0.5% activity with raucaffricine compared to the wild type enzyme Rauvolfia serpentina
E476A the mutant shows 0.57% activity with raucaffricine compared to the wild type enzyme Rauvolfia serpentina
E476L the mutant shows 0.67% activity with raucaffricine compared to the wild type enzyme Rauvolfia serpentina
F485Y the mutant shows 24.24% activity with raucaffricine compared to the wild type enzyme Rauvolfia serpentina
H193A the mutant shows 43.66% activity with raucaffricine compared to the wild type enzyme Rauvolfia serpentina
S390G the mutant shows 30.91% activity with raucaffricine compared to the wild type enzyme Rauvolfia serpentina
T189A the mutant shows 63.94% activity with raucaffricine compared to the wild type enzyme Rauvolfia serpentina
W392A the mutant shows 1.21% activity with raucaffricine compared to the wild type enzyme Rauvolfia serpentina
Y200A the mutant shows 4.6% activity with raucaffricine compared to the wild type enzyme Rauvolfia serpentina

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.3
-
Raucaffricine wild type enzyme, pH and temperature not specified in the publication Rauvolfia serpentina
1.8
-
strictosidine wild type enzyme, pH and temperature not specified in the publication Rauvolfia serpentina

Organism

Organism UniProt Comment Textmining
Rauvolfia serpentina
-
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1,2-(S)-dihydro-raucaffricine + H2O 98% acivity compared to raucaffricine Rauvolfia serpentina 2beta(R)-1,2-dihydrovomilenine + D-glucose
-
?
additional information no activity with arbutin Rauvolfia serpentina ?
-
?
raucaffricine + H2O 100% activity Rauvolfia serpentina D-glucose + vomilenine
-
?
secologanin + H2O 0.24% acivity compared to raucaffricine Rauvolfia serpentina secologanin aglycone
-
?
strictosidine + H2O 1.18% acivity compared to raucaffricine Rauvolfia serpentina strictosidine aglycone + D-glucose
-
?

Synonyms

Synonyms Comment Organism
raucaffricine glucosidase
-
Rauvolfia serpentina
raucaffricine O-beta-D-glucosidase
-
Rauvolfia serpentina