Crystallization (Comment) | Organism |
---|---|
the crystal structure of mFMO that contains both cofactors, FAD and NADP+, is analyzed | Methylophaga sp. |
Protein Variants | Comment | Organism |
---|---|---|
C78A | site-directed mutagenesis, the mutation leads to an increase in KM and kcat compared to wild-type enzyme, but the mutant shows reduced activity compared to wild-type enzyme | Methylophaga sp. |
C78I | site-directed mutagenesis, the mutation leads to an increase in KM and kcat compared to wild-type enzyme, but the mutant shows reduced activity compared to wild-type enzyme | Methylophaga sp. |
C78L | site-directed mutagenesis, the mutant shows reduced activity compared to wild-type enzyme | Methylophaga sp. |
C78V | site-directed mutagenesis, the mutation leads to an increase in KM and kcat compared to wild-type enzyme, but the mutant shows reduced activity compared to wild-type enzyme | Methylophaga sp. |
M15L/S23A | site-directed mutagenesis, combining the two mutations at the N-terminus results in a 3°C increase in apparent melting temperature. Both M15L and S23A are far from the active site and no significant effect on the kinetic parameters of the enzyme is observed. Adding more stabilizing mutations does not contribute to a higher thermostability | Methylophaga sp. |
W319A | site-directed mutagenesis, the mutant shows reduced activity compared to wild-type enzyme | Methylophaga sp. |
W319F | site-directed mutagenesis, the mutant shows reduced activity compared to wild-type enzyme | Methylophaga sp. |
Y207W | site-directed mutagenesis, the mutant shows reduced activity compared to wild-type enzyme | Methylophaga sp. |
Y207W | site-directed mutagenesis, the mutation leads to an increase in KM and kcat, as well as in catalytic efficiency, compared to wild-type enzyme | Methylophaga sp. |
Y207W/W319A | site-directed mutagenesis, the mutant shows reduced activity compared to wild-type enzyme | Methylophaga sp. |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | steady-state kinetic analyses, overview | Methylophaga sp. | |
0.1 | - |
indole | pH 8.0, 25°C, recombinant Y207W | Methylophaga sp. | |
0.4 | - |
indole | pH 8.0, 25°C, recombinant wild-type enzyme | Methylophaga sp. | |
0.4 | - |
indole | pH 8.0, 25°C, recombinant C78A | Methylophaga sp. | |
0.4 | - |
indole | pH 8.0, 25°C, recombinant W319F | Methylophaga sp. | |
0.7 | - |
indole | pH 8.0, 25°C, recombinant C78V | Methylophaga sp. | |
0.8 | - |
indole | pH 8.0, 25°C, recombinant C78I | Methylophaga sp. | |
0.8 | - |
indole | pH 8.0, 25°C, recombinant C78I/Y207W/W319A | Methylophaga sp. | |
0.8 | - |
indole | pH 8.0, 25°C, recombinant C78L | Methylophaga sp. | |
0.8 | - |
indole | pH 8.0, 25°C, recombinant Y207W/W319A | Methylophaga sp. | |
0.9 | - |
indole | pH 8.0, 25°C, recombinant W319A | Methylophaga sp. |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
NaCl | the enzyme is optimally active at 250 mM NaCl, with an activity increase of more than 25% compared to the enzyme in the absence of NaCl | Methylophaga sp. |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
indole + NADPH + H+ + O2 | Methylophaga sp. | - |
indole N-oxide + NADP+ + H2O | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Methylophaga sp. | - |
- |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
indole + NADPH + H+ + O2 | - |
Methylophaga sp. | indole N-oxide + NADP+ + H2O | - |
? |
Synonyms | Comment | Organism |
---|---|---|
flavin-containing monooxygenase | - |
Methylophaga sp. |
FMO | - |
Methylophaga sp. |
mFMO | - |
Methylophaga sp. |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Methylophaga sp. |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
43.3 | - |
enzyme mFMO falls into the category of moderately stable enzymes with an apparent melting temperature of 43.3°C | Methylophaga sp. |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.31 | - |
indole | pH 8.0, 25°C, recombinant Y207W | Methylophaga sp. | |
0.42 | - |
indole | pH 8.0, 25°C, recombinant W319A | Methylophaga sp. | |
0.64 | - |
indole | pH 8.0, 25°C, recombinant W319F | Methylophaga sp. | |
0.7 | - |
indole | pH 8.0, 25°C, recombinant C78L | Methylophaga sp. | |
0.79 | - |
indole | pH 8.0, 25°C, recombinant C78A | Methylophaga sp. | |
0.8 | - |
indole | pH 8.0, 25°C, recombinant Y207W/W319A | Methylophaga sp. | |
0.85 | - |
indole | pH 8.0, 25°C, recombinant wild-type enzyme | Methylophaga sp. | |
0.93 | - |
indole | pH 8.0, 25°C, recombinant C78I/Y207W/W319A | Methylophaga sp. | |
1.04 | - |
indole | pH 8.0, 25°C, recombinant C78V | Methylophaga sp. | |
1.28 | - |
indole | pH 8.0, 25°C, recombinant C78I | Methylophaga sp. |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
- |
Methylophaga sp. |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
7 | 9.5 | over 50% of maximal activity within this range | Methylophaga sp. |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FAD | - |
Methylophaga sp. | |
NADPH | - |
Methylophaga sp. |
General Information | Comment | Organism |
---|---|---|
malfunction | a sidechain at position Gly74 can have interactions with the substrate. Despite being a pre-Pro residue, the phi-psi angles (-130,175°) will also fit non-Glycine residues. Mutation of Cys78 can fill up a cavity in the active site making binding of indole more productive. Tyr207 and Asp317 form part of the entrance to the substrate binding cavity while residues Trp319, Phe397, and Trp400 limit the size of the substrate binding cavity | Methylophaga sp. |
additional information | enzyme structure and active site structure analysis using the structure with PDB ID 2VQ7, overview | Methylophaga sp. |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.47 | - |
indole | pH 8.0, 25°C, recombinant W319A | Methylophaga sp. | |
0.875 | - |
indole | pH 8.0, 25°C, recombinant C78L | Methylophaga sp. | |
1 | - |
indole | pH 8.0, 25°C, recombinant Y207W/W319A | Methylophaga sp. | |
1.163 | - |
indole | pH 8.0, 25°C, recombinant C78I/Y207W/W319A | Methylophaga sp. | |
1.5 | - |
indole | pH 8.0, 25°C, recombinant C78V | Methylophaga sp. | |
1.6 | - |
indole | pH 8.0, 25°C, recombinant C78I | Methylophaga sp. | |
1.6 | - |
indole | pH 8.0, 25°C, recombinant W319F | Methylophaga sp. | |
1.975 | - |
indole | pH 8.0, 25°C, recombinant C78A | Methylophaga sp. | |
2.125 | - |
indole | pH 8.0, 25°C, recombinant wild-type enzyme | Methylophaga sp. | |
3.1 | - |
indole | pH 8.0, 25°C, recombinant Y207W | Methylophaga sp. |