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Literature summary for 1.11.2.1 extracted from

  • Hayakawa, S.; Matsumura, H.; Nakamura, N.; Yohda, M.; Ohno, H.
    Identification of the rate-limiting step of the peroxygenase reactions catalyzed by the thermophilic cytochrome P450 from Sulfolobus tokodaii strain 7 (2014), FEBS J., 281, 1409-1416 .
    View publication on PubMed

Protein Variants

Protein Variants Comment Organism
F310A/A320Q site-directed mutagenesis, the mutant exhibits a 30 mV positive shift in redox potential for the FeIII/FeII couple compared with wild-type P450st due to weakening of the electron-donating effect (push effect) of the proximal thiolate in the mutant. This result indicates that the electron density around the heme is decreased, and thus the Lewis acidity of the heme is expected to be increased. Mutant F310A/A320Q maintains higher thermal stability than typical P450s Sulfurisphaera tokodaii

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
23.2
-
ethylbenzene pH 5.0, 25°C Sulfurisphaera tokodaii
26.2
-
ethylbenzene pH 10.0, 25°C Sulfurisphaera tokodaii
81.4
-
ethylbenzene pH 7.0, 25°C Sulfurisphaera tokodaii

Metals/Ions

Metals/Ions Comment Organism Structure
Iron a FeIII/FeII couple in the heme cofactor of cytochrome P-450 Sulfurisphaera tokodaii

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ethylbenzene + H2O2 Sulfurisphaera tokodaii
-
1-phenylethanol + H2O
-
?
ethylbenzene + H2O2 Sulfurisphaera tokodaii 7
-
1-phenylethanol + H2O
-
?

Organism

Organism UniProt Comment Textmining
Sulfurisphaera tokodaii Q972I2
-
-
Sulfurisphaera tokodaii 7 Q972I2
-
-

Reaction

Reaction Comment Organism Reaction ID
RH + H2O2 = ROH + H2O kinetic mechanism, overview Sulfurisphaera tokodaii

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ethylbenzene + H2O2
-
Sulfurisphaera tokodaii 1-phenylethanol + H2O
-
?
ethylbenzene + H2O2
-
Sulfurisphaera tokodaii 7 1-phenylethanol + H2O
-
?
additional information the enzyme catalyzes hydrogen peroxide-driven ethylbenzene hydroxylation, as well as styrene epoxidation. The ethylbenzene hydroxylation activity is higher than the styrene epoxidation activity, maybe due to a difference in the binding affinity of the two substrates. The rate-limiting steps of ethylbenzene hydroxylation and styrene epoxidation are the same, and may be any step before formation of the active oxidant Sulfurisphaera tokodaii ?
-
?
additional information the enzyme catalyzes hydrogen peroxide-driven ethylbenzene hydroxylation, as well as styrene epoxidation. The ethylbenzene hydroxylation activity is higher than the styrene epoxidation activity, maybe due to a difference in the binding affinity of the two substrates. The rate-limiting steps of ethylbenzene hydroxylation and styrene epoxidation are the same, and may be any step before formation of the active oxidant Sulfurisphaera tokodaii 7 ?
-
?

Synonyms

Synonyms Comment Organism
P450st
-
Sulfurisphaera tokodaii

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
assay at Sulfurisphaera tokodaii

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.0093
-
ethylbenzene pH 5.0, 25°C Sulfurisphaera tokodaii
0.125
-
ethylbenzene pH 7.0, 25°C Sulfurisphaera tokodaii
0.158
-
ethylbenzene pH 10.0, 25°C Sulfurisphaera tokodaii

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
5
-
first optimum Sulfurisphaera tokodaii
10
-
second optimum Sulfurisphaera tokodaii

pH Range

pH Minimum pH Maximum Comment Organism
3 12 activity range, profile overview. Analysis of pH dependencies of wild-type and mutant enzymes Sulfurisphaera tokodaii

Cofactor

Cofactor Comment Organism Structure
cytochrome P450
-
Sulfurisphaera tokodaii
heme containing a FeIII/FeII couple Sulfurisphaera tokodaii
NADH
-
Sulfurisphaera tokodaii

General Information

General Information Comment Organism
malfunction the F310A/A320Q mutant catalyzes styrene epoxidation via the peroxide shunt pathway more efficiently than wild-type P450st in neutral solutions Sulfurisphaera tokodaii
metabolism styrene epoxidation via the peroxide shunt pathway Sulfurisphaera tokodaii
physiological function cytochrome P450 from the thermoacidophilic crenarchaeon Sulfolobus tokodaii strain 7 (P450st) is a thermophilic cytochrome P450 that shows high tolerance of harsh conditions and is capable of catalyzing some peroxygenase reactions. Both hydrogen peroxide-driven ethylbenzene hydroxylation and styrene epoxidation by wild-type P450st are found to be activated in weak acidic and weak basic solutions Sulfurisphaera tokodaii

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.0004
-
ethylbenzene pH 5.0, 25°C Sulfurisphaera tokodaii
0.0015
-
ethylbenzene pH 7.0, 25°C Sulfurisphaera tokodaii
0.006
-
ethylbenzene pH 10.0, 25°C Sulfurisphaera tokodaii