BRENDA - Enzyme Database show
show all sequences of 1.14.14.36

Purification and characterization of recombinant cytochrome P450tyr expressed at high levels in Escherichia coli

Halkier, B.A.; Nielsen, H.L.; Koch, B.; Moller B.L.; Arch. Biochem. Biophys. 322, 369-377 (1995)

Data extracted from this reference:

Activating Compound
Activating Compound
Commentary
Organism
Structure
glutathione
3 mM glutathione stimulates activity of the reconstituted enzyme; at 3 mM, activation rate differs between experiments
Sorghum bicolor
Cloned(Commentary)
Commentary
Organism
expressed in Escherichia coli JM109 cells; various N-terminal modifications
Sorghum bicolor
Engineering
Amino acid exchange
Commentary
Organism
additional information
mutant 1: first codons of Escherichia coli mRNA are enriched for A's and T's, second codon is changed into GCT, first 8 codons of P450 sequence are replaced with the N-terminal sequence of bovine P45017alpha, mutant 2: deletion of 14 amino acids, mutant 3: deletion of 25 amino acids, mutant 4: deletion of 75 amino acids
Sorghum bicolor
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.22
-
L-tyrosine
in reconstitution experiments using Sorghum bicolor; recombinant enzyme, in 50 mM Tricine, pH 7.9, a 30°C
Sorghum bicolor
Localization
Localization
Commentary
Organism
GeneOntology No.
Textmining
membrane
-
Sorghum bicolor
16020
-
microsome
-
Sorghum bicolor
-
-
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
61700
-
calculated from amino acid sequence; SDS-PAGE
Sorghum bicolor
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
L-tyrosine + O2 + [reduced NADPH-hemoprotein reductase]
Sorghum bicolor
enzyme in biosynthesis of cyanogenic glucosides
N-hydroxy-L-tyrosine + [oxidized NADPH-hemoprotein reductase] + H2O
-
-
-
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Sorghum bicolor
-
-
-
Purification (Commentary)
Commentary
Organism
DEAE-Speharose column chromatography and Reactive Red-120 agarose column chromatography; homogeneity
Sorghum bicolor
Specific Activity [micromol/min/mg]
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
0.0233
-
-
Sorghum bicolor
49.2
-
-
Sorghum bicolor
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
L-tyrosine + 2 O2 + 2 [reduced NADPH-hemoprotein reductase]
-
390080
Sorghum bicolor
(E)-[4-hydroxyphenylacetaldehyde oxime] + 2 [oxidized NADPH-hemoprotein reductase] + CO2 + 3 H2O
-
390080
Sorghum bicolor
?
L-tyrosine + 2 O2 + 2 [reduced NADPH-hemoprotein reductase]
overall reaction, P450Tyr is a multifunctional tyrosine N-hydroxylase catalyzing the double N-hydroxylation of L-tyrosine to N,N-dihydroxy-L-tyrosine which dehydrates and decarboxylates to 4-hydroxyphenylacetaldoxime
390080
Sorghum bicolor
(E)-[4-hydroxyphenylacetaldehyde oxime] + 2 [oxidized NADPH-hemoprotein reductase] + CO2 + 3 H2O
-
-
-
?
L-tyrosine + O2 + [reduced NADPH-hemoprotein reductase]
enzyme in biosynthesis of cyanogenic glucosides
390080
Sorghum bicolor
N-hydroxy-L-tyrosine + [oxidized NADPH-hemoprotein reductase] + H2O
-
-
-
-
additional information
P450Tyr does not metabolize L-phenylalanine
390080
Sorghum bicolor
?
-
-
-
-
N-hydroxy-L-tyrosine + 2 O2 + 2 [reduced NADPH-hemoprotein reductase]
-
390080
Sorghum bicolor
N,N-dihydroxy-L-tyrosine + [oxidized NADPH-hemoprotein reductase] + H2O
-
-
-
?
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.82
-
L-tyrosine
in reconstitution experiments using Sorghum bicolor; purified recombinant enzyme, in 50 mM Tricine, pH 7.9, a 30°C
Sorghum bicolor
5.83
-
L-tyrosine
in E. coli membranes; unpurified recombinant enzyme in Escherichia coli membranes, in 50 mM Tricine, pH 7.9, a 30°C
Sorghum bicolor
Cofactor
Cofactor
Commentary
Organism
Structure
NADPH
-
Sorghum bicolor
Activating Compound (protein specific)
Activating Compound
Commentary
Organism
Structure
glutathione
3 mM glutathione stimulates activity of the reconstituted enzyme; at 3 mM, activation rate differs between experiments
Sorghum bicolor
Cloned(Commentary) (protein specific)
Commentary
Organism
expressed in Escherichia coli JM109 cells; various N-terminal modifications
Sorghum bicolor
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
NADPH
-
Sorghum bicolor
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
additional information
mutant 1: first codons of Escherichia coli mRNA are enriched for A's and T's, second codon is changed into GCT, first 8 codons of P450 sequence are replaced with the N-terminal sequence of bovine P45017alpha, mutant 2: deletion of 14 amino acids, mutant 3: deletion of 25 amino acids, mutant 4: deletion of 75 amino acids
Sorghum bicolor
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.22
-
L-tyrosine
in reconstitution experiments using Sorghum bicolor; recombinant enzyme, in 50 mM Tricine, pH 7.9, a 30°C
Sorghum bicolor
Localization (protein specific)
Localization
Commentary
Organism
GeneOntology No.
Textmining
membrane
-
Sorghum bicolor
16020
-
microsome
-
Sorghum bicolor
-
-
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
61700
-
calculated from amino acid sequence; SDS-PAGE
Sorghum bicolor
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
L-tyrosine + O2 + [reduced NADPH-hemoprotein reductase]
Sorghum bicolor
enzyme in biosynthesis of cyanogenic glucosides
N-hydroxy-L-tyrosine + [oxidized NADPH-hemoprotein reductase] + H2O
-
-
-
Purification (Commentary) (protein specific)
Commentary
Organism
DEAE-Speharose column chromatography and Reactive Red-120 agarose column chromatography; homogeneity
Sorghum bicolor
Specific Activity [micromol/min/mg] (protein specific)
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
0.0233
-
-
Sorghum bicolor
49.2
-
-
Sorghum bicolor
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
L-tyrosine + 2 O2 + 2 [reduced NADPH-hemoprotein reductase]
-
390080
Sorghum bicolor
(E)-[4-hydroxyphenylacetaldehyde oxime] + 2 [oxidized NADPH-hemoprotein reductase] + CO2 + 3 H2O
-
390080
Sorghum bicolor
?
L-tyrosine + 2 O2 + 2 [reduced NADPH-hemoprotein reductase]
overall reaction, P450Tyr is a multifunctional tyrosine N-hydroxylase catalyzing the double N-hydroxylation of L-tyrosine to N,N-dihydroxy-L-tyrosine which dehydrates and decarboxylates to 4-hydroxyphenylacetaldoxime
390080
Sorghum bicolor
(E)-[4-hydroxyphenylacetaldehyde oxime] + 2 [oxidized NADPH-hemoprotein reductase] + CO2 + 3 H2O
-
-
-
?
L-tyrosine + O2 + [reduced NADPH-hemoprotein reductase]
enzyme in biosynthesis of cyanogenic glucosides
390080
Sorghum bicolor
N-hydroxy-L-tyrosine + [oxidized NADPH-hemoprotein reductase] + H2O
-
-
-
-
additional information
P450Tyr does not metabolize L-phenylalanine
390080
Sorghum bicolor
?
-
-
-
-
N-hydroxy-L-tyrosine + 2 O2 + 2 [reduced NADPH-hemoprotein reductase]
-
390080
Sorghum bicolor
N,N-dihydroxy-L-tyrosine + [oxidized NADPH-hemoprotein reductase] + H2O
-
-
-
?
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.82
-
L-tyrosine
in reconstitution experiments using Sorghum bicolor; purified recombinant enzyme, in 50 mM Tricine, pH 7.9, a 30°C
Sorghum bicolor
5.83
-
L-tyrosine
in E. coli membranes; unpurified recombinant enzyme in Escherichia coli membranes, in 50 mM Tricine, pH 7.9, a 30°C
Sorghum bicolor
Other publictions for EC 1.14.14.36
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)
736574
Gnanasekaran
Transfer of the cytochrome P45 ...
Sorghum bicolor
J. Exp. Bot.
67
2495-2506
2016
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1
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739696
Bavishi
Application of nanodisc techno ...
Sorghum bicolor
Sci. Rep.
6
29459
2016
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1
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4
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1
1
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739318
Clausen
The bifurcation of the cyanoge ...
Sorghum bicolor
Plant J.
84
558-573
2015
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1
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1
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4
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3
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4
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1
1
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660383
Kristensen
Metabolic engineering of dhurr ...
Sorghum bicolor
Proc. Natl. Acad. Sci. USA
102
1779-1784
2005
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1
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6
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660217
Busk
Dhurrin synthesis in sorghum i ...
Sorghum bicolor
Plant Physiol.
129
1222-1231
2002
-
-
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3
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2
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2
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1
1
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739340
Nielsen
Cloning and expression of cyto ...
Triglochin maritima
Plant Physiol.
122
1311-1321
2000
-
-
1
-
-
-
-
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6
-
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4
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4
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1
2
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739341
Bak
Transgenic tobacco and Arabido ...
Sorghum bicolor
Plant Physiol.
123
1437-1448
2000
-
1
-
-
-
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-
-
-
-
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5
-
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390081
Kahn
Substrate specificity of the c ...
Sorghum bicolor
Arch. Biochem. Biophys.
363
9-18
1999
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2
1
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1
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4
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1
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4
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1
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4
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390082
Bak
The presence of CYP79 homologu ...
Sinapis alba
Plant Mol. Biol.
38
725-734
1998
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11
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739339
Kahn
Isolation and reconstitution o ...
Sorghum bicolor
Plant Physiol.
115
1661-1670
1997
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390079
Koch
The primary sequence of cytoch ...
Sorghum bicolor
Arch. Biochem. Biophys.
323
177-186
1995
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6
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390080
Halkier
Purification and characterizat ...
Sorghum bicolor
Arch. Biochem. Biophys.
322
369-377
1995
1
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1
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1
2
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1
1
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5
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738563
Sibbesen
Cytochrome P-450TYR is a multi ...
Sorghum bicolor
J. Biol. Chem.
270
3506-3511
1995
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390078
Libbesen
Isolation of the heme-thiolate ...
Sorghum bicolor
Proc. Natl. Acad. Sci. USA
91
9740-9744
1994
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348459
Halkier
The biosynthesis of cyanogenic ...
Sorghum bicolor
J. Biol. Chem.
265
21114-21121
1990
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6
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6
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285331
McFarlane
Conn, E.E.: The in vitro biosy ...
Sorghum bicolor
J. Biol. Chem.
250
4708-4713
1975
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