BRENDA - Enzyme Database show
show all sequences of 1.14.14.90

Functional expression of cytochrome P450 in Escherichia coli An approach to functional analysis of uncharacterized enzymes for flavonoid biosynthesis

Uchida, K.; Akashi, T.; Aoki, T.; Plant Biotechnol. 32, 205-213 (2015)
No PubMed abstract available

Data extracted from this reference:

Activating Compound
Activating Compound
Commentary
Organism
Structure
protein CPR
is an essential redox partner for functional expression of CYP; is an essential redox partner for functional expression of CYP
Glycine max
Cloned(Commentary)
Commentary
Organism
dual plasmid method development for functional expression of plant CYPs in Escherichia coli, method and culture conditions optimization, overview. Gene CYP81E11, DNA and amino acid sequence determination and analysis, recombinant expression of the transmembrane-domain truncated CYP enzyme in Escherichia coli strain C41(DE3) and coexpression with CPR from Lotus japonicus as a discrete polypeptide. The optimal temperature is 25°C, addition of the heme precursor 5-aminolevulinic acid is essential for functional expression of CYP81E11; dual plasmid method development for functional expression of plant CYPs in Escherichia coli, method and culture conditions optimiztaion, overview. Gene CYP81E12, DNA and amino acid sequence determination and analysis, recombinant expression of the transmembrane-domain truncated CYP enzyme in Escherichia coli strain C41(DE3) and coexpression with CPR from Lotus japonicus as a discrete polypeptide. The optimal temperature is 25°C, addition of the heme precursor 5-aminolevulinic acid is essential for functional expression of CYP81E12; dual plasmid method development for functional expression of plant CYPs in Escherichia coli, method and culture conditions optimiztaion, overview. Gene CYP81E13, DNA and amino acid sequence determination and analysis, recombinant expression of the transmembrane-domain truncated CYP enzyme in Escherichia coli strain C41(DE3) and coexpression with CPR from Lotus japonicus as a discrete polypeptide. The optimal temperature is 25°C, addition of the heme precursor 5-aminolevulinic acid is essential for functional expression of CYP81E13
Glycine max
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
daidzein + [reduced NADPH-hemoprotein reductase] + O2
Glycine max
-
2'-hydroxydaidzein + [oxidized NADPH-hemoprotein reductase] + H2O
-
-
?
genistein + [reduced NADPH-hemoprotein reductase] + O2
Glycine max
-
2'-hydroxygenistein + [oxidized NADPH-hemoprotein reductase] + H2O
-
-
?
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Glycine max
-
cv. Tamba Kurodaizu
-
Glycine max
Q2LAL0
cv. Tamba Kurodaizu
-
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
daidzein + [reduced NADPH-hemoprotein reductase] + O2
-
746010
Glycine max
2'-hydroxydaidzein + [oxidized NADPH-hemoprotein reductase] + H2O
-
-
-
?
genistein + [reduced NADPH-hemoprotein reductase] + O2
-
746010
Glycine max
2'-hydroxygenistein + [oxidized NADPH-hemoprotein reductase] + H2O
-
-
-
?
moe
no activity with formononetin, cf. EC 1.14.13.53, 4'-methoxyisoflavone 2'-hydroxylase
746010
Glycine max
?
-
-
-
-
Cofactor
Cofactor
Commentary
Organism
Structure
cytochrome P-450
;
Glycine max
heme
;
Glycine max
NADPH-hemoprotein reductase
A flavoprotein containing both FMN and FAD. This enzyme catalyses the transfer of electrons from NADPH, an obligatory two-electron donor, to microsomal P-450 monooxygenases, EC 1.14.14._; A flavoprotein containing both FMN and FAD. This enzyme catalyses the transfer of electrons from NADPH, an obligatory two-electron donor, to microsomal P-450 monooxygenases, EC 1.14.14._
Glycine max
Activating Compound (protein specific)
Activating Compound
Commentary
Organism
Structure
protein CPR
is an essential redox partner for functional expression of CYP
Glycine max
Cloned(Commentary) (protein specific)
Commentary
Organism
dual plasmid method development for functional expression of plant CYPs in Escherichia coli, method and culture conditions optimiztaion, overview. Gene CYP81E12, DNA and amino acid sequence determination and analysis, recombinant expression of the transmembrane-domain truncated CYP enzyme in Escherichia coli strain C41(DE3) and coexpression with CPR from Lotus japonicus as a discrete polypeptide. The optimal temperature is 25°C, addition of the heme precursor 5-aminolevulinic acid is essential for functional expression of CYP81E12; dual plasmid method development for functional expression of plant CYPs in Escherichia coli, method and culture conditions optimiztaion, overview. Gene CYP81E13, DNA and amino acid sequence determination and analysis, recombinant expression of the transmembrane-domain truncated CYP enzyme in Escherichia coli strain C41(DE3) and coexpression with CPR from Lotus japonicus as a discrete polypeptide. The optimal temperature is 25°C, addition of the heme precursor 5-aminolevulinic acid is essential for functional expression of CYP81E13
Glycine max
dual plasmid method development for functional expression of plant CYPs in Escherichia coli, method and culture conditions optimization, overview. Gene CYP81E11, DNA and amino acid sequence determination and analysis, recombinant expression of the transmembrane-domain truncated CYP enzyme in Escherichia coli strain C41(DE3) and coexpression with CPR from Lotus japonicus as a discrete polypeptide. The optimal temperature is 25°C, addition of the heme precursor 5-aminolevulinic acid is essential for functional expression of CYP81E11
Glycine max
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
cytochrome P-450
-
Glycine max
heme
-
Glycine max
NADPH-hemoprotein reductase
A flavoprotein containing both FMN and FAD. This enzyme catalyses the transfer of electrons from NADPH, an obligatory two-electron donor, to microsomal P-450 monooxygenases, EC 1.14.14._
Glycine max
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
daidzein + [reduced NADPH-hemoprotein reductase] + O2
Glycine max
-
2'-hydroxydaidzein + [oxidized NADPH-hemoprotein reductase] + H2O
-
-
?
genistein + [reduced NADPH-hemoprotein reductase] + O2
Glycine max
-
2'-hydroxygenistein + [oxidized NADPH-hemoprotein reductase] + H2O
-
-
?
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
daidzein + [reduced NADPH-hemoprotein reductase] + O2
-
746010
Glycine max
2'-hydroxydaidzein + [oxidized NADPH-hemoprotein reductase] + H2O
-
-
-
?
genistein + [reduced NADPH-hemoprotein reductase] + O2
-
746010
Glycine max
2'-hydroxygenistein + [oxidized NADPH-hemoprotein reductase] + H2O
-
-
-
?
moe
no activity with formononetin, cf. EC 1.14.13.53, 4'-methoxyisoflavone 2'-hydroxylase
746010
Glycine max
?
-
-
-
-
Other publictions for EC 1.14.14.90
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)
746010
Uchida
-
Functional expression of cyto ...
Glycine max
Plant Biotechnol.
32
205-213
2015
1
-
1
-
-
-
-
-
-
-
-
4
-
2
-
-
-
-
-
-
-
-
6
-
-
-
-
-
-
-
-
3
-
-
-
2
-
2
6
-
-
-
-
-
-
-
-
-
-
4
-
-
-
-
-
-
-
-
6
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
746367
Chen
Cloning, expression and purif ...
Astragalus membranaceus
Protein Expr. Purif.
107
83-89
2015
-
-
1
-
1
-
-
-
1
-
-
-
-
4
-
-
1
-
-
1
-
-
-
1
-
-
-
-
-
-
-
2
-
-
-
-
-
1
2
-
1
-
-
-
-
-
1
-
-
-
-
-
-
1
-
1
-
-
-
1
-
-
-
-
-
-
-
-
-
2
2
-
-
-
660173
Liu
Regiospecific hydroxylation of ...
Medicago truncatula
Plant J.
36
471-484
2003
-
-
1
-
-
-
-
2
1
-
-
-
-
6
-
-
-
-
-
1
-
-
6
-
-
-
-
2
1
-
-
1
-
-
-
-
-
1
1
-
-
-
-
-
-
2
1
-
-
-
-
-
-
-
-
1
-
-
6
-
-
-
-
2
1
-
-
-
-
-
-
-
-
-
285324
Shimada
Induction of isoflavonoid path ...
Lotus japonicus
Plant Sci.
160
37-47
2000
-
-
1
-
-
-
-
-
-
-
-
1
-
4
-
-
-
-
-
1
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
1
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
658293
Nakamura
Induction of isoflavonoid and ...
Glycyrrhiza echinata, Glycyrrhiza echinata AK-1
Biosci. Biotechnol. Biochem.
63
1618-1620
1999
-
-
1
-
-
-
-
-
-
-
-
1
-
4
-
-
-
-
-
1
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
1
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
285323
Akashi
CYP81E1, a cytochrome P450 cDN ...
Glycyrrhiza echinata, Glycyrrhiza echinata L.
Biochem. Biophys. Res. Commun.
251
67-70
1998
-
-
1
-
-
-
-
-
-
-
-
1
-
4
-
-
-
-
-
-
-
-
5
-
-
-
-
-
-
-
-
1
-
-
-
-
-
1
1
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
5
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-