BRENDA - Enzyme Database show
show all sequences of 1.14.19.66

Molecular cloning and heterologous expression of a cDNA encoding berbamunine synthase, a C-O phenol-coupling cytochrome P450 from the higher plant Berberis stolonifera

Kraus, P.F.X.; Kutchan, T.M.; Proc. Natl. Acad. Sci. USA 92, 2071-2075 (1995)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
expression in functional form in insect cell culture using a baculovirus-based expression system; expression in Sf9 cells
Berberis stolonifera
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
additional information
-
additional information
normal Michaelis-Menten kinetics are not observed for the formation of guattegaumerine or berbamunine
Berberis stolonifera
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
46000
-
x * 46000, SDS-PAGE
Berberis stolonifera
55474
-
x * 55474, calculation from nucleotide sequence
Berberis stolonifera
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Berberis stolonifera
P47195
-
-
Purification (Commentary)
Commentary
Organism
heterologously expressed enzyme from Sf9 cell culture
Berberis stolonifera
Source Tissue
Source Tissue
Commentary
Organism
Textmining
cell suspension culture
-
Berberis stolonifera
-
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2 (S)-N-methylcoclaurine + 2 (R)-N-methylcoclaurine + [reduced NADPH-hemoprotein reductase] + O2
-
393213
Berberis stolonifera
berbamunine + guattegaumerine + [oxidized NADPH-hemoprotein reductase] + H2O
two molecules of (R)-N-methylcoclaurine for the (R,R) dimer guattegaumerine or one molecule each of (R)-N-methylcoclaurine and (S)-N-methylcoclaurine form the (R,S) dimer berbamunine. The ratio of the two bisbenzylisoquinolines formed can be altered by the reductase source or by varying the enantiomer composition of the substrates
393213
Berberis stolonifera
-
Subunits
Subunits
Commentary
Organism
?
x * 46000, SDS-PAGE; x * 55474, calculation from nucleotide sequence
Berberis stolonifera
Temperature Optimum [°C]
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
35
-
formation of berbamunine
Berberis stolonifera
38
-
formation of guattegaumerine
Berberis stolonifera
Temperature Range [°C]
Temperature Minimum [°C]
Temperature Maximum [°C]
Commentary
Organism
7
49
50% of maximal activity at 7°C and 49°C, formation of berbamunine
Berberis stolonifera
18
46
50% of maximal activity at 18°C and 46°C, formation of guattegaumerine
Berberis stolonifera
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
additional information
-
additional information
-
Berberis stolonifera
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.5
-
formation of berbamunine or guattegaumerine
Berberis stolonifera
Cloned(Commentary) (protein specific)
Commentary
Organism
expression in functional form in insect cell culture using a baculovirus-based expression system; expression in Sf9 cells
Berberis stolonifera
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
additional information
-
additional information
normal Michaelis-Menten kinetics are not observed for the formation of guattegaumerine or berbamunine
Berberis stolonifera
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
46000
-
x * 46000, SDS-PAGE
Berberis stolonifera
55474
-
x * 55474, calculation from nucleotide sequence
Berberis stolonifera
Purification (Commentary) (protein specific)
Commentary
Organism
heterologously expressed enzyme from Sf9 cell culture
Berberis stolonifera
Source Tissue (protein specific)
Source Tissue
Commentary
Organism
Textmining
cell suspension culture
-
Berberis stolonifera
-
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2 (S)-N-methylcoclaurine + 2 (R)-N-methylcoclaurine + [reduced NADPH-hemoprotein reductase] + O2
-
393213
Berberis stolonifera
berbamunine + guattegaumerine + [oxidized NADPH-hemoprotein reductase] + H2O
two molecules of (R)-N-methylcoclaurine for the (R,R) dimer guattegaumerine or one molecule each of (R)-N-methylcoclaurine and (S)-N-methylcoclaurine form the (R,S) dimer berbamunine. The ratio of the two bisbenzylisoquinolines formed can be altered by the reductase source or by varying the enantiomer composition of the substrates
393213
Berberis stolonifera
-
Subunits (protein specific)
Subunits
Commentary
Organism
?
x * 46000, SDS-PAGE; x * 55474, calculation from nucleotide sequence
Berberis stolonifera
Temperature Optimum [°C] (protein specific)
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
35
-
formation of berbamunine
Berberis stolonifera
38
-
formation of guattegaumerine
Berberis stolonifera
Temperature Range [°C] (protein specific)
Temperature Minimum [°C]
Temperature Maximum [°C]
Commentary
Organism
7
49
50% of maximal activity at 7°C and 49°C, formation of berbamunine
Berberis stolonifera
18
46
50% of maximal activity at 18°C and 46°C, formation of guattegaumerine
Berberis stolonifera
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
additional information
-
additional information
-
Berberis stolonifera
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.5
-
formation of berbamunine or guattegaumerine
Berberis stolonifera
Other publictions for EC 1.14.19.66
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
393214
Kutchan
Heterologous expression of alk ...
Berberis stolonifera
Gene
179
73-81
1996
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-
1
-
-
-
-
-
-
-
-
-
-
4
-
-
1
-
-
-
1
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
1
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
393213
Kraus
Molecular cloning and heterolo ...
Berberis stolonifera
Proc. Natl. Acad. Sci. USA
92
2071-2075
1995
-
-
1
-
-
-
-
1
-
-
2
-
-
3
-
-
1
-
-
3
-
-
1
1
2
2
-
1
1
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
1
-
-
2
-
-
-
-
1
-
3
-
-
1
1
2
2
-
1
1
-
-
-
-
-
-
-
-
-
393212
Stadler
The purification and character ...
Berberis stolonifera
J. Biol. Chem.
268
823-831
1993
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-
-
-
-
-
-
-
-
-
1
1
-
1
-
-
1
1
-
1
1
-
4
1
1
1
-
1
1
-
-
1
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
1
1
-
-
-
1
-
1
1
-
4
1
1
1
-
1
1
-
-
-
-
-
-
-
-
-