BRENDA - Enzyme Database

beta-Amyrin synthase from Euphorbia tirucalli L. functional analyses of the highly conserved aromatic residues Phe413, Tyr259 and Trp257 disclose the importance of the appropriate steric bulk, and cation-Pi and CH-Pi interactions for the efficient catalytic

Ito, R.; Nakada, C.; Hoshino, T.; Org. Biomol. Chem. 15, 177-188 (2017)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Cloned (Commentary)
Organism
5.4.99.39
gene EtAS, sequence comparisons, recombinant expression of wild-type and mutant enzymes in yeast
Euphorbia tirucalli
Engineering
EC Number
Protein Variants
Commentary
Organism
5.4.99.39
F413A
site-directed mutagenesis, functional analysis, the mutant shows slightly reduced activity compared to thre wild-type enzyme
Euphorbia tirucalli
5.4.99.39
F413H
site-directed mutagenesis, functional analysis, the mutant shows slightly increased activity compared to thre wild-type enzyme
Euphorbia tirucalli
5.4.99.39
F413M
site-directed mutagenesis, functional analysis, the mutant shows slightly reduced activity compared to thre wild-type enzyme
Euphorbia tirucalli
5.4.99.39
F413S
site-directed mutagenesis, functional analysis, the mutant shows slightly increased activity compared to thre wild-type enzyme
Euphorbia tirucalli
5.4.99.39
F413T
site-directed mutagenesis, functional analysis, the mutant shows slightly increased activity compared to thre wild-type enzyme
Euphorbia tirucalli
5.4.99.39
F413V
site-directed mutagenesis, functional analysis, the mutant shows slightly reduced activity compared to thre wild-type enzyme
Euphorbia tirucalli
5.4.99.39
F413W
site-directed mutagenesis, functional analysis, the mutant shows slightly increased activity compared to thre wild-type enzyme
Euphorbia tirucalli
5.4.99.39
F413Y
site-directed mutagenesis, functional analysis, the mutant shows slightly reduced activity compared to thre wild-type enzyme
Euphorbia tirucalli
5.4.99.39
additional information
quantities of oleanane-type, 17-epi-dammarane-type, and dammarane-type triterpenes in cultures of wild-type and mutant strains differ significantly, overview
Euphorbia tirucalli
5.4.99.39
Y259A
site-directed mutagenesis, functional analysis
Euphorbia tirucalli
5.4.99.39
Y259F
site-directed mutagenesis, functional analysis
Euphorbia tirucalli
5.4.99.39
Y259H
site-directed mutagenesis, functional analysis
Euphorbia tirucalli
5.4.99.39
Y259L
site-directed mutagenesis, functional analysis
Euphorbia tirucalli
5.4.99.39
Y259V
site-directed mutagenesis, functional analysis
Euphorbia tirucalli
5.4.99.39
Y259W
site-directed mutagenesis, functional analysis
Euphorbia tirucalli
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
5.4.99.39
(3S)-2,3-epoxy-2,3-dihydrosqualene
Euphorbia tirucalli
-
beta-amyrin
-
-
r
Organism
EC Number
Organism
UniProt
Commentary
Textmining
5.4.99.39
Euphorbia tirucalli
Q401R6
-
-
Reaction
EC Number
Reaction
Commentary
Organism
Reaction ID
5.4.99.39
(3S)-2,3-epoxy-2,3-dihydrosqualene = beta-amyrin
the Phe416 residue is located near the D-ring formation site and works to position the oxidosqualene substrate correctly within the reaction cavity. On the other hand, the major catalysis-related function of the Tyr259 and Trp257 residues is to yield their Pi-electrons to the cationic intermediate
Euphorbia tirucalli
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
5.4.99.39
(3S)-2,3-epoxy-2,3-dihydrosqualene
-
748812
Euphorbia tirucalli
beta-amyrin
-
-
-
r
5.4.99.39
additional information
product identification by EIMS and NMR spectral analyses
748812
Euphorbia tirucalli
?
-
-
-
-
Synonyms
EC Number
Synonyms
Commentary
Organism
5.4.99.39
EtAS
-
Euphorbia tirucalli
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
5.4.99.39
gene EtAS, sequence comparisons, recombinant expression of wild-type and mutant enzymes in yeast
Euphorbia tirucalli
Engineering (protein specific)
EC Number
Protein Variants
Commentary
Organism
5.4.99.39
F413A
site-directed mutagenesis, functional analysis, the mutant shows slightly reduced activity compared to thre wild-type enzyme
Euphorbia tirucalli
5.4.99.39
F413H
site-directed mutagenesis, functional analysis, the mutant shows slightly increased activity compared to thre wild-type enzyme
Euphorbia tirucalli
5.4.99.39
F413M
site-directed mutagenesis, functional analysis, the mutant shows slightly reduced activity compared to thre wild-type enzyme
Euphorbia tirucalli
5.4.99.39
F413S
site-directed mutagenesis, functional analysis, the mutant shows slightly increased activity compared to thre wild-type enzyme
Euphorbia tirucalli
5.4.99.39
F413T
site-directed mutagenesis, functional analysis, the mutant shows slightly increased activity compared to thre wild-type enzyme
Euphorbia tirucalli
5.4.99.39
F413V
site-directed mutagenesis, functional analysis, the mutant shows slightly reduced activity compared to thre wild-type enzyme
Euphorbia tirucalli
5.4.99.39
F413W
site-directed mutagenesis, functional analysis, the mutant shows slightly increased activity compared to thre wild-type enzyme
Euphorbia tirucalli
5.4.99.39
F413Y
site-directed mutagenesis, functional analysis, the mutant shows slightly reduced activity compared to thre wild-type enzyme
Euphorbia tirucalli
5.4.99.39
additional information
quantities of oleanane-type, 17-epi-dammarane-type, and dammarane-type triterpenes in cultures of wild-type and mutant strains differ significantly, overview
Euphorbia tirucalli
5.4.99.39
Y259A
site-directed mutagenesis, functional analysis
Euphorbia tirucalli
5.4.99.39
Y259F
site-directed mutagenesis, functional analysis
Euphorbia tirucalli
5.4.99.39
Y259H
site-directed mutagenesis, functional analysis
Euphorbia tirucalli
5.4.99.39
Y259L
site-directed mutagenesis, functional analysis
Euphorbia tirucalli
5.4.99.39
Y259V
site-directed mutagenesis, functional analysis
Euphorbia tirucalli
5.4.99.39
Y259W
site-directed mutagenesis, functional analysis
Euphorbia tirucalli
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
5.4.99.39
(3S)-2,3-epoxy-2,3-dihydrosqualene
Euphorbia tirucalli
-
beta-amyrin
-
-
r
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
5.4.99.39
(3S)-2,3-epoxy-2,3-dihydrosqualene
-
748812
Euphorbia tirucalli
beta-amyrin
-
-
-
r
5.4.99.39
additional information
product identification by EIMS and NMR spectral analyses
748812
Euphorbia tirucalli
?
-
-
-
-
General Information
EC Number
General Information
Commentary
Organism
5.4.99.39
malfunction
the Y259F variant shows nearly equivalent activity to that of the wild type, but aliphatic mutants such as the Ala, Val, and Leu variants show significantly decreased activity and yield the tetracyclic dammarane scaffold. The aliphatic variants of Trp257 exhibit remarkably decreased enzymatic activity, and lupeol is produced in a high production ratio. The aromatic Phe and Tyr mutants exhibit high activities owing to their more increased Pi-electron density relative to that of the aliphatic mutants, but lupeol is produced in a significantly high yield besides beta-amyrin. The Trp residue is likely to be responsible for the robust binding of Me-30 through CH-Pi interaction. The decreased Pi-electron density of the Phe and Tyr mutants compared to that of Trp results in the high production of lupeol
Euphorbia tirucalli
5.4.99.39
additional information
the highly conserved aromatic residues Phe413, Tyr259 and Trp257 disclose the importance of the appropriate steric bulk, and cation-Pi and CH-Pi interactions for the efficient catalytic action of the polyolefin cyclization cascade, functional analysis, overview. Homology modeling of beta-amyrin synthase using the X-ray crystal structure of human lanosterol cyclase, PDB ID 1w6k, as template. Structure comparisons. The Tyr259 residue stabilizes the baccharenyl secondary cation via cation-Pi interaction, residue Trp257 stabilizes the oleanyl cation via cation-Pi interaction
Euphorbia tirucalli
General Information (protein specific)
EC Number
General Information
Commentary
Organism
5.4.99.39
malfunction
the Y259F variant shows nearly equivalent activity to that of the wild type, but aliphatic mutants such as the Ala, Val, and Leu variants show significantly decreased activity and yield the tetracyclic dammarane scaffold. The aliphatic variants of Trp257 exhibit remarkably decreased enzymatic activity, and lupeol is produced in a high production ratio. The aromatic Phe and Tyr mutants exhibit high activities owing to their more increased Pi-electron density relative to that of the aliphatic mutants, but lupeol is produced in a significantly high yield besides beta-amyrin. The Trp residue is likely to be responsible for the robust binding of Me-30 through CH-Pi interaction. The decreased Pi-electron density of the Phe and Tyr mutants compared to that of Trp results in the high production of lupeol
Euphorbia tirucalli
5.4.99.39
additional information
the highly conserved aromatic residues Phe413, Tyr259 and Trp257 disclose the importance of the appropriate steric bulk, and cation-Pi and CH-Pi interactions for the efficient catalytic action of the polyolefin cyclization cascade, functional analysis, overview. Homology modeling of beta-amyrin synthase using the X-ray crystal structure of human lanosterol cyclase, PDB ID 1w6k, as template. Structure comparisons. The Tyr259 residue stabilizes the baccharenyl secondary cation via cation-Pi interaction, residue Trp257 stabilizes the oleanyl cation via cation-Pi interaction
Euphorbia tirucalli