Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21(DE3) cells | Methylovorus sp. MP688 |
Crystallization (Comment) | Organism |
---|---|
hanging drop vapor diffusion method, using 16-22% (w/v) PEG3350 and 200 mM magnesium formate | Methylovorus sp. MP688 |
Protein Variants | Comment | Organism |
---|---|---|
F434A | the mutant shows reduced activity with 4-hydroxy-3-methoxybenzyl alcohol compared to the wild type enzyme | Methylovorus sp. MP688 |
H467A | the mutant shows reduced activity with 4-hydroxy-3-methoxybenzyl alcohol compared to the wild type enzyme | Methylovorus sp. MP688 |
M103A | the mutant shows reduced activity with 4-hydroxy-3-methoxybenzyl alcohol compared to the wild type enzyme | Methylovorus sp. MP688 |
N511A | the mutant shows reduced activity with 4-hydroxy-3-methoxybenzyl alcohol compared to the wild type enzyme | Methylovorus sp. MP688 |
V367 | the mutant shows increased activity with 5-formylfuran-2-carboxylate compared to the wild type enzyme | Methylovorus sp. MP688 |
V367K | the mutant shows reduced activity with 4-hydroxy-3-methoxybenzyl alcohol compared to the wild type enzyme | Methylovorus sp. MP688 |
V367R/W466F | the mutant shows strongly increased activity with 5-formylfuran-2-carboxylate compared to the wild type enzyme | Methylovorus sp. MP688 |
V465A | the mutant shows reduced activity with 4-hydroxy-3-methoxybenzyl alcohol compared to the wild type enzyme | Methylovorus sp. MP688 |
W369A | the mutant shows reduced activity with 4-hydroxy-3-methoxybenzyl alcohol compared to the wild type enzyme | Methylovorus sp. MP688 |
W466A | the mutant shows reduced activity with 4-hydroxy-3-methoxybenzyl alcohol compared to the wild type enzyme | Methylovorus sp. MP688 |
W466F | the mutant shows increased activity with 5-formylfuran-2-carboxylate compared to the wild type enzyme | Methylovorus sp. MP688 |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.2 | - |
4-Hydroxy-3-methoxybenzyl alcohol | mutant enzyme W466F, at pH 7.5 and 25°C | Methylovorus sp. MP688 | |
0.21 | - |
5-formylfuran-2-carboxylate | mutant enzyme V367R/W466F, at pH 7.5 and 25°C | Methylovorus sp. MP688 | |
0.28 | - |
4-Hydroxy-3-methoxybenzyl alcohol | mutant enzyme V367R, at pH 7.5 and 25°C | Methylovorus sp. MP688 | |
0.29 | - |
4-Hydroxy-3-methoxybenzyl alcohol | mutant enzyme V367K, at pH 7.5 and 25°C | Methylovorus sp. MP688 | |
0.72 | - |
4-Hydroxy-3-methoxybenzyl alcohol | wild type enzyme, at pH 7.5 and 25°C | Methylovorus sp. MP688 | |
0.82 | - |
4-Hydroxy-3-methoxybenzyl alcohol | mutant enzyme H467A, at pH 7.5 and 25°C | Methylovorus sp. MP688 | |
0.92 | - |
4-Hydroxy-3-methoxybenzyl alcohol | mutant enzyme V465A, at pH 7.5 and 25°C | Methylovorus sp. MP688 | |
1 | - |
4-Hydroxy-3-methoxybenzyl alcohol | mutant enzyme W466A, at pH 7.5 and 25°C | Methylovorus sp. MP688 | |
1.1 | - |
4-Hydroxy-3-methoxybenzyl alcohol | mutant enzyme W369A, at pH 7.5 and 25°C | Methylovorus sp. MP688 | |
1.2 | - |
4-Hydroxy-3-methoxybenzyl alcohol | mutant enzyme F434A, at pH 7.5 and 25°C | Methylovorus sp. MP688 | |
1.4 | - |
5-formylfuran-2-carboxylate | mutant enzyme V367R, at pH 7.5 and 25°C | Methylovorus sp. MP688 | |
1.8 | - |
4-Hydroxy-3-methoxybenzyl alcohol | mutant enzyme M103A, at pH 7.5 and 25°C | Methylovorus sp. MP688 | |
2 | - |
4-Hydroxy-3-methoxybenzyl alcohol | mutant enzyme N511A, at pH 7.5 and 25°C | Methylovorus sp. MP688 | |
3 | - |
5-formylfuran-2-carboxylate | mutant enzyme W466F, at pH 7.5 and 25°C | Methylovorus sp. MP688 | |
4 | - |
5-formylfuran-2-carboxylate | Km above 4.0 mM, wild type enzyme, at pH 7.5 and 25°C | Methylovorus sp. MP688 | |
20 | - |
(S)-1-phenylethanol | mutant enzyme W466A, at pH 7.5 and 25°C | Methylovorus sp. MP688 | |
98 | - |
(S)-1-phenylethanol | mutant enzyme W466F, at pH 7.5 and 25°C | Methylovorus sp. MP688 |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
5-(dihydroxymethyl)furan-2-carbaldehyde + O2 | Methylovorus sp. MP688 | - |
5-formylfuran-2-carboxylate + H2O2 | - |
? | |
5-(dihydroxymethyl)furan-2-carboxylate + O2 | Methylovorus sp. MP688 | - |
furan-2,5-dicarboxylate + H2O2 | - |
? | |
5-(hydroxymethyl)furfural + 3 O2 + 2 H2O | Methylovorus sp. MP688 | overall reaction | furan-2,5-dicarboxylate + 3 H2O2 | - |
? | |
5-(hydroxymethyl)furfural + O2 | Methylovorus sp. MP688 | - |
furan-2,5-dicarbaldehyde + H2O2 | - |
? | |
5-formylfuran-2-carboxylate + H2O | Methylovorus sp. MP688 | - |
5-(dihydroxymethyl)furan-2-carboxylate | - |
? | |
furan-2,5-dicarbaldehyde + H2O | Methylovorus sp. MP688 | - |
5-(dihydroxymethyl)furan-2-carbaldehyde | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Methylovorus sp. MP688 | E4QP00 | - |
- |
Purification (Comment) | Organism |
---|---|
HisTrap column chromatography and Superdex 200 gel filtration | Methylovorus sp. MP688 |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(R)-1-phenylethanol + O2 | - |
Methylovorus sp. MP688 | acetophenone + H2O2 | - |
? | |
(S)-1-phenylethanol + O2 | - |
Methylovorus sp. MP688 | acetophenone + H2O2 | - |
? | |
4-hydroxy-3-methoxybenzyl alcohol + O2 | - |
Methylovorus sp. MP688 | 4-hydroxy-3-methoxybenzaldehyde + H2O2 | - |
? | |
5-(dihydroxymethyl)furan-2-carbaldehyde + O2 | - |
Methylovorus sp. MP688 | 5-formylfuran-2-carboxylate + H2O2 | - |
? | |
5-(dihydroxymethyl)furan-2-carboxylate + O2 | - |
Methylovorus sp. MP688 | furan-2,5-dicarboxylate + H2O2 | - |
? | |
5-(hydroxymethyl)furfural + 3 O2 + 2 H2O | overall reaction | Methylovorus sp. MP688 | furan-2,5-dicarboxylate + 3 H2O2 | - |
? | |
5-(hydroxymethyl)furfural + O2 | - |
Methylovorus sp. MP688 | furan-2,5-dicarbaldehyde + H2O2 | - |
? | |
5-formylfuran-2-carboxylate + H2O | - |
Methylovorus sp. MP688 | 5-(dihydroxymethyl)furan-2-carboxylate | - |
? | |
furan-2,5-dicarbaldehyde + H2O | - |
Methylovorus sp. MP688 | 5-(dihydroxymethyl)furan-2-carbaldehyde | - |
? |
Synonyms | Comment | Organism |
---|---|---|
5-hydroxymethylfurfural oxidase | - |
Methylovorus sp. MP688 |
HMFO | - |
Methylovorus sp. MP688 |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0047 | - |
4-Hydroxy-3-methoxybenzyl alcohol | mutant enzyme H467A, at pH 7.5 and 25°C | Methylovorus sp. MP688 | |
0.01 | - |
(S)-1-phenylethanol | mutant enzyme W466F, at pH 7.5 and 25°C | Methylovorus sp. MP688 | |
0.011 | - |
(S)-1-phenylethanol | mutant enzyme W466A, at pH 7.5 and 25°C | Methylovorus sp. MP688 | |
0.056 | - |
5-formylfuran-2-carboxylate | mutant enzyme V367R, at pH 7.5 and 25°C | Methylovorus sp. MP688 | |
0.079 | - |
5-formylfuran-2-carboxylate | mutant enzyme W466F, at pH 7.5 and 25°C | Methylovorus sp. MP688 | |
0.46 | - |
5-formylfuran-2-carboxylate | mutant enzyme V367R/W466F, at pH 7.5 and 25°C | Methylovorus sp. MP688 | |
0.75 | - |
4-Hydroxy-3-methoxybenzyl alcohol | mutant enzyme W466A, at pH 7.5 and 25°C | Methylovorus sp. MP688 | |
1 | - |
4-Hydroxy-3-methoxybenzyl alcohol | mutant enzyme N511A, at pH 7.5 and 25°C | Methylovorus sp. MP688 | |
1.5 | - |
4-Hydroxy-3-methoxybenzyl alcohol | mutant enzyme V465A, at pH 7.5 and 25°C | Methylovorus sp. MP688 | |
1.7 | - |
4-Hydroxy-3-methoxybenzyl alcohol | mutant enzyme W466F, at pH 7.5 and 25°C | Methylovorus sp. MP688 | |
3.3 | - |
4-Hydroxy-3-methoxybenzyl alcohol | mutant enzyme M103A, at pH 7.5 and 25°C | Methylovorus sp. MP688 | |
4.3 | - |
4-Hydroxy-3-methoxybenzyl alcohol | mutant enzyme W369A, at pH 7.5 and 25°C | Methylovorus sp. MP688 | |
5.1 | - |
4-Hydroxy-3-methoxybenzyl alcohol | mutant enzyme V367K, at pH 7.5 and 25°C | Methylovorus sp. MP688 | |
6 | - |
4-Hydroxy-3-methoxybenzyl alcohol | mutant enzyme V367R, at pH 7.5 and 25°C | Methylovorus sp. MP688 | |
15 | - |
4-Hydroxy-3-methoxybenzyl alcohol | mutant enzyme F434A, at pH 7.5 and 25°C | Methylovorus sp. MP688 | |
21 | - |
4-Hydroxy-3-methoxybenzyl alcohol | wild type enzyme, at pH 7.5 and 25°C | Methylovorus sp. MP688 |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FAD | - |
Methylovorus sp. MP688 |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0001 | - |
(S)-1-phenylethanol | mutant enzyme W466F, at pH 7.5 and 25°C | Methylovorus sp. MP688 | |
0.00055 | - |
(S)-1-phenylethanol | mutant enzyme W466A, at pH 7.5 and 25°C | Methylovorus sp. MP688 | |
0.005 | - |
5-formylfuran-2-carboxylate | wild type enzyme, at pH 7.5 and 25°C | Methylovorus sp. MP688 | |
0.0057 | - |
4-Hydroxy-3-methoxybenzyl alcohol | mutant enzyme H467A, at pH 7.5 and 25°C | Methylovorus sp. MP688 | |
0.026 | - |
5-formylfuran-2-carboxylate | mutant enzyme W466F, at pH 7.5 and 25°C | Methylovorus sp. MP688 | |
0.041 | - |
5-formylfuran-2-carboxylate | mutant enzyme V367R, at pH 7.5 and 25°C | Methylovorus sp. MP688 | |
0.55 | - |
4-Hydroxy-3-methoxybenzyl alcohol | mutant enzyme N511A, at pH 7.5 and 25°C | Methylovorus sp. MP688 | |
0.75 | - |
4-Hydroxy-3-methoxybenzyl alcohol | mutant enzyme W466A, at pH 7.5 and 25°C | Methylovorus sp. MP688 | |
1.6 | - |
4-Hydroxy-3-methoxybenzyl alcohol | mutant enzyme V465A, at pH 7.5 and 25°C | Methylovorus sp. MP688 | |
1.8 | - |
4-Hydroxy-3-methoxybenzyl alcohol | mutant enzyme M103A, at pH 7.5 and 25°C | Methylovorus sp. MP688 | |
2.2 | - |
5-formylfuran-2-carboxylate | mutant enzyme V367R/W466F, at pH 7.5 and 25°C | Methylovorus sp. MP688 | |
3.9 | - |
4-Hydroxy-3-methoxybenzyl alcohol | mutant enzyme W369A, at pH 7.5 and 25°C | Methylovorus sp. MP688 | |
8.5 | - |
4-Hydroxy-3-methoxybenzyl alcohol | mutant enzyme W466F, at pH 7.5 and 25°C | Methylovorus sp. MP688 | |
12 | - |
4-Hydroxy-3-methoxybenzyl alcohol | mutant enzyme F434A, at pH 7.5 and 25°C | Methylovorus sp. MP688 | |
18 | - |
4-Hydroxy-3-methoxybenzyl alcohol | mutant enzyme V367K, at pH 7.5 and 25°C | Methylovorus sp. MP688 | |
21 | - |
4-Hydroxy-3-methoxybenzyl alcohol | mutant enzyme V367R, at pH 7.5 and 25°C | Methylovorus sp. MP688 | |
29 | - |
4-Hydroxy-3-methoxybenzyl alcohol | wild type enzyme, at pH 7.5 and 25°C | Methylovorus sp. MP688 |