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Literature summary for 1.6.2.4 extracted from

  • Sarapusit, S.; Pethuan, S.; Rongnoparut, P.
    Mosquito NADPH-cytochrome P450 oxidoreductase: kinetics and role of phenylalanine amino acid substitutions at leu86 and leu219 in CYP6AA3-mediated deltamethrin metabolism (2010), Arch. Insect Biochem. Physiol., 73, 232-244.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli XL-1 Blue cells Anopheles minimus

Protein Variants

Protein Variants Comment Organism
L219F the mutant upon reconstitution with the Anopheles minimus cytochrome P450 CYP6AA3 and a NADPH-regenerating system, increases CYP6AA3-mediated deltamethrin degradation compared to the wild type enzyme Anopheles minimus
L86F the mutant upon reconstitution with the Anopheles minimus cytochrome P450 CYP6AA3 and a NADPH-regenerating system, increases CYP6AA3-mediated deltamethrin degradation compared to the wild type enzyme Anopheles minimus
L86F/L219F the mutant upon reconstitution with the Anopheles minimus cytochrome P450 CYP6AA3 and a NADPH-regenerating system, increases CYP6AA3-mediated deltamethrin degradation compared to the wild type enzyme Anopheles minimus
additional information DELTA55AnCYPOR has an approximately 0.5fold increased FAD content compared to the wild type enzyme Anopheles minimus

Inhibitors

Inhibitors Comment Organism Structure
NADP+ competitive inhibition Anopheles minimus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0116
-
NADPH mutant enzyme L86F/L219F, pH not specified in the publication, at 25°C Anopheles minimus
0.0125
-
NADPH wild type enzyme, pH not specified in the publication, at 25°C Anopheles minimus
0.019
-
ferricytochrome c wild type enzyme, pH not specified in the publication, at 25°C Anopheles minimus
0.0239
-
ferricytochrome c mutant enzyme L86F/L219F, pH not specified in the publication, at 25°C Anopheles minimus
4
-
NADH wild type enzyme, pH not specified in the publication, at 25°C Anopheles minimus
4.3
-
NADH mutant enzyme L86F/L219F, pH not specified in the publication, at 25°C Anopheles minimus

Localization

Localization Comment Organism GeneOntology No. Textmining
membrane
-
Anopheles minimus 16020
-

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
83000
-
His-tagged enzyme, SDS-PAGE Anopheles minimus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2 ferricytochrome P450 + NADPH Anopheles minimus
-
2 ferrocytochrome P450 + NADP+ + H+
-
?

Organism

Organism UniProt Comment Textmining
Anopheles minimus
-
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni-NTA column chromatography and Superdex G25 gel filtration, the purified enzyme readily loses its flavin cofactors Anopheles minimus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2 ferricytochrome c + NADH lower binding affinity for NADH compared to NADPH Anopheles minimus 2 ferrocytochrome c + NAD+ + H+
-
?
2 ferricytochrome c + NADPH
-
Anopheles minimus 2 ferrocytochrome c + NADP+ + H+
-
?
2 ferricytochrome P450 + NADPH
-
Anopheles minimus 2 ferrocytochrome P450 + NADP+ + H+
-
?

Synonyms

Synonyms Comment Organism
CYPOR
-
Anopheles minimus
NADPH-cytochrome P450 oxidoreductase
-
Anopheles minimus

Cofactor

Cofactor Comment Organism Structure
FAD the enzyme contains 0.22 mol of FAD per mol of protein, when supplemented with exogenous flavin cofactors, the activity of purified CYPOR-mediated cytochrome c reduction is increased Anopheles minimus
FMN the enzyme contains 0.11 mol of FMN per mol of protein, when supplemented with exogenous flavin cofactors, the activity of purified CYPOR-mediated cytochrome c reduction is increased Anopheles minimus
NADPH
-
Anopheles minimus

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.0115
-
NADP+ wild type enzyme, pH not specified in the publication, at 25°C Anopheles minimus