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Literature summary extracted from

  • Han, A.; Robinson, R.M.; Badieyan, S.; Ellerbrock, J.; Sobrado, P.
    Tryptophan-47 in the active site of Methylophaga sp. strain SK1 flavin-monooxygenase is important for hydride transfer (2013), Arch. Biochem. Biophys., 532, 46-53.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.14.13.8 expression in Escherichia coli Methylophaga sp.

Protein Variants

EC Number Protein Variants Comment Organism
1.14.13.8 W47A insoluble inactive protein Methylophaga sp.
1.14.13.8 W47F soluble and active protein. The spectrum of the flavin displays a redshift, the kcat values for NADPH, trimethylamine, and methimazole, show a 5-8fold decrease, and primary kinetic isotope effect values are higher than in wild-type. Mutant displays reduced flexibility in active site residues and, in particular, the nicotinamide moiety of NADP+ Methylophaga sp.

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.14.13.8 0.0015
-
 trimethylamine mutant W47F, pH 8.5, 25°C Methylophaga sp.
1.14.13.8 0.0026
-
 trimethylamine wild-type, pH 8.5, 25°C Methylophaga sp.
1.14.13.8 0.0041
-
 NADPH wild-type, pH 8.5, 25°C Methylophaga sp.
1.14.13.8 0.0043
-
 NADPH mutant W47F, pH 8.5, 25°C Methylophaga sp.
1.14.13.8 0.019
-
 O2 wild-type, pH 8.5, 25°C Methylophaga sp.
1.14.13.8 0.037
-
 Methimazole mutant W47F, pH 8.5, 25°C Methylophaga sp.
1.14.13.8 0.042
-
 O2 mutant W47F, pH 8.5, 25°C Methylophaga sp.
1.14.13.8 0.163
-
 Methimazole wild-type, pH 8.5, 25°C Methylophaga sp.

Organism

EC Number Organism UniProt Comment Textmining
1.14.13.8 Methylophaga sp.
-
-
-
1.14.13.8 Methylophaga sp. SK1
-
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.14.13.8 methimazole + NADPH + H+ + O2
-
 Methylophaga sp. methimazole N-oxide + NADP+ + H2O
-
 ?
1.14.13.8 methimazole + NADPH + H+ + O2
-
 Methylophaga sp. SK1 methimazole N-oxide + NADP+ + H2O
-
 ?
1.14.13.8 trimethylamine + NADPH + H+ + O2
-
 Methylophaga sp. trimethylamine N-oxide + NADP+ + H2O
-
 ?
1.14.13.8 trimethylamine + NADPH + H+ + O2
-
 Methylophaga sp. SK1 trimethylamine N-oxide + NADP+ + H2O
-
 ?

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.14.13.8 0.67
-
 Methimazole mutant W47F, pH 8.5, 25°C Methylophaga sp.
1.14.13.8 0.92
-
 NADPH mutant W47F, pH 8.5, 25°C Methylophaga sp.
1.14.13.8 1.25
-
 trimethylamine mutant W47F, pH 8.5, 25°C Methylophaga sp.
1.14.13.8 2.7
-
 Methimazole wild-type, pH 8.5, 25°C Methylophaga sp.
1.14.13.8 2.7
-
 O2 mutant W47F, pH 8.5, 25°C Methylophaga sp.
1.14.13.8 6.1
-
 trimethylamine wild-type, pH 8.5, 25°C Methylophaga sp.
1.14.13.8 7.3
-
 NADPH wild-type, pH 8.5, 25°C Methylophaga sp.
1.14.13.8 7.7
-
 O2 wild-type, pH 8.5, 25°C Methylophaga sp.

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.14.13.8 16
-
 Methimazole wild-type, pH 8.5, 25°C Methylophaga sp.
1.14.13.8 18
-
 Methimazole mutant W47F, pH 8.5, 25°C Methylophaga sp.
1.14.13.8 60
-
 O2 mutant W47F, pH 8.5, 25°C Methylophaga sp.
1.14.13.8 210
-
 NADPH mutant W47F, pH 8.5, 25°C Methylophaga sp.
1.14.13.8 410
-
 O2 wild-type, pH 8.5, 25°C Methylophaga sp.
1.14.13.8 800
-
 trimethylamine mutant W47F, pH 8.5, 25°C Methylophaga sp.
1.14.13.8 1800
-
 NADPH wild-type, pH 8.5, 25°C Methylophaga sp.
1.14.13.8 2300
-
 trimethylamine wild-type, pH 8.5, 25°C Methylophaga sp.