BRENDA - Enzyme Database
show all sequences of 1.14.13.B28

Monooxygenation by a thermophilic cytochrome P450 via direct electron donation from NADH

Matsumura, H.; Matsuda, K.; Nakamura, N.; Ohtaki, A.; Yoshida, H.; Kamitori, S.; Yohda, M.; Ohno, H.; Metallomics 3, 389-395 (2011)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
wild-type enzyme and F-G loop deletion mutant enzyme delLL151-E156, overexpression in Escherichia coli
Sulfurisphaera tokodaii
Crystallization (Commentary)
Crystallization
Organism
vapour diffusion method, X-ray crystallography at a resolution of 1.94 A reveals a sufficiently large heme pocket for NAD(P)H binding and a novel contiguous channel from the active site to bulk solvent in the distal heme pocket. The mutant shows a higher affinity for NADH compared with the wild-type because the mutant has a more widely open distal pocket for NAD(P)H binding
Sulfurisphaera tokodaii
Engineering
Amino acid exchange
Commentary
Organism
delL151-E156
the Km value of the mutant is about 2times lower than that of the wild-type.
Sulfurisphaera tokodaii
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.29
-
Styrene
pH 7, 25°C, wild-type enzyme
Sulfurisphaera tokodaii
0.52
-
Styrene
pH 7, 25°C, mutant enzyme delLL151-E156
Sulfurisphaera tokodaii
7
-
NADH
pH 7, 25°C, mutant enzyme delLL151-E156
Sulfurisphaera tokodaii
13
-
NADH
pH 7, 25°C, wild-type enzyme
Sulfurisphaera tokodaii
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
43000
-
F-G loop deletion mutant enzyme delLL151-E156
Sulfurisphaera tokodaii
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Sulfurisphaera tokodaii
Q972I2
-
-
Sulfurisphaera tokodaii 7
Q972I2
-
-
Purification (Commentary)
Commentary
Organism
-
Sulfurisphaera tokodaii
Reaction
Reaction
Commentary
Organism
styrene + NADH + H+ + O2 = styrene epoxide + NAD+ + H2O
sequential mechanism. Both styrene and NADH bind to the enzyme before any product is released
Sulfurisphaera tokodaii
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
styrene + NADH + H+ + O2
the initial rate of catalysis with NADH is slightly higher than with NADPH
725874
Sulfurisphaera tokodaii
styrene epoxide + NAD+ + H2O
-
-
-
?
styrene + NADH + H+ + O2
the initial rate of catalysis with NADH is slightly higher than with NADPH
725874
Sulfurisphaera tokodaii 7
styrene epoxide + NAD+ + H2O
-
-
-
?
styrene + NADPH + H+ + O2
the initial rate of catalysis with NADH is slightly higher than with NADPH
725874
Sulfurisphaera tokodaii
styrene epoxide + NADP+ + H2O
-
-
-
?
styrene + NADPH + H+ + O2
the initial rate of catalysis with NADH is slightly higher than with NADPH
725874
Sulfurisphaera tokodaii 7
styrene epoxide + NADP+ + H2O
-
-
-
?
Temperature Optimum [°C]
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
25
-
assay at
Sulfurisphaera tokodaii
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.000057
-
Styrene
pH 7, 25°C, mutant enzyme delLL151-E156
Sulfurisphaera tokodaii
0.000061
-
Styrene
pH 7, 25°C, wild-type enzyme
Sulfurisphaera tokodaii
0.000076
-
NADH
pH 7, 25°C, wild-type enzyme
Sulfurisphaera tokodaii
0.000079
-
NADH
pH 7, 25°C, mutant enzyme delLL151-E156
Sulfurisphaera tokodaii
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7
-
assay at
Sulfurisphaera tokodaii
Cofactor
Cofactor
Commentary
Organism
Structure
NADH
the initial rate of catalysis with NADH is slightly higher than with NADPH
Sulfurisphaera tokodaii
NADPH
the initial rate of catalysis with NADH is slightly higher than with NADPH
Sulfurisphaera tokodaii
Cloned(Commentary) (protein specific)
Commentary
Organism
wild-type enzyme and F-G loop deletion mutant enzyme delLL151-E156, overexpression in Escherichia coli
Sulfurisphaera tokodaii
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
NADH
the initial rate of catalysis with NADH is slightly higher than with NADPH
Sulfurisphaera tokodaii
NADPH
the initial rate of catalysis with NADH is slightly higher than with NADPH
Sulfurisphaera tokodaii
Crystallization (Commentary) (protein specific)
Crystallization
Organism
vapour diffusion method, X-ray crystallography at a resolution of 1.94 A reveals a sufficiently large heme pocket for NAD(P)H binding and a novel contiguous channel from the active site to bulk solvent in the distal heme pocket. The mutant shows a higher affinity for NADH compared with the wild-type because the mutant has a more widely open distal pocket for NAD(P)H binding
Sulfurisphaera tokodaii
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
delL151-E156
the Km value of the mutant is about 2times lower than that of the wild-type.
Sulfurisphaera tokodaii
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.29
-
Styrene
pH 7, 25°C, wild-type enzyme
Sulfurisphaera tokodaii
0.52
-
Styrene
pH 7, 25°C, mutant enzyme delLL151-E156
Sulfurisphaera tokodaii
7
-
NADH
pH 7, 25°C, mutant enzyme delLL151-E156
Sulfurisphaera tokodaii
13
-
NADH
pH 7, 25°C, wild-type enzyme
Sulfurisphaera tokodaii
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
43000
-
F-G loop deletion mutant enzyme delLL151-E156
Sulfurisphaera tokodaii
Purification (Commentary) (protein specific)
Commentary
Organism
-
Sulfurisphaera tokodaii
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
styrene + NADH + H+ + O2
the initial rate of catalysis with NADH is slightly higher than with NADPH
725874
Sulfurisphaera tokodaii
styrene epoxide + NAD+ + H2O
-
-
-
?
styrene + NADH + H+ + O2
the initial rate of catalysis with NADH is slightly higher than with NADPH
725874
Sulfurisphaera tokodaii 7
styrene epoxide + NAD+ + H2O
-
-
-
?
styrene + NADPH + H+ + O2
the initial rate of catalysis with NADH is slightly higher than with NADPH
725874
Sulfurisphaera tokodaii
styrene epoxide + NADP+ + H2O
-
-
-
?
styrene + NADPH + H+ + O2
the initial rate of catalysis with NADH is slightly higher than with NADPH
725874
Sulfurisphaera tokodaii 7
styrene epoxide + NADP+ + H2O
-
-
-
?
Temperature Optimum [°C] (protein specific)
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
25
-
assay at
Sulfurisphaera tokodaii
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.000057
-
Styrene
pH 7, 25°C, mutant enzyme delLL151-E156
Sulfurisphaera tokodaii
0.000061
-
Styrene
pH 7, 25°C, wild-type enzyme
Sulfurisphaera tokodaii
0.000076
-
NADH
pH 7, 25°C, wild-type enzyme
Sulfurisphaera tokodaii
0.000079
-
NADH
pH 7, 25°C, mutant enzyme delLL151-E156
Sulfurisphaera tokodaii
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7
-
assay at
Sulfurisphaera tokodaii
Other publictions for EC 1.14.13.B28
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)
744870
Hayakawa
Identification of the rate-li ...
Sulfurisphaera tokodaii, Sulfurisphaera tokodaii 7
FEBS J.
281
1409-1416
2014
-
-
-
-
1
-
-
-
-
1
-
2
-
4
-
-
-
-
-
-
-
-
4
-
1
-
-
1
2
1
-
3
-
-
-
-
-
-
3
-
1
-
-
-
-
-
-
1
-
2
-
-
-
-
-
-
-
-
4
-
1
-
-
1
2
1
-
-
-
3
3
-
-
-
725874
Matsumura
Monooxygenation by a thermophi ...
Sulfurisphaera tokodaii, Sulfurisphaera tokodaii 7
Metallomics
3
389-395
2011
-
-
1
1
1
-
-
4
-
-
1
-
-
4
-
-
1
1
-
-
-
-
4
-
1
-
-
4
1
-
-
2
-
-
-
-
-
1
2
1
1
-
-
-
-
4
-
-
1
-
-
-
-
1
-
-
-
-
4
-
1
-
-
4
1
-
-
-
-
-
-
-
-
-
748359
Oku
Structure and direct electroc ...
Sulfurisphaera tokodaii, Sulfurisphaera tokodaii 7
J. Inorg. Biochem.
98
1194-1199
2004
-
-
1
1
-
-
-
-
-
-
-
-
-
4
-
-
1
-
-
-
-
-
2
1
-
1
1
-
-
-
-
1
-
-
-
-
-
1
1
1
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
2
1
-
1
1
-
-
-
-
-
-
-
-
-
-
-