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Literature summary extracted from

  • Dijkman, W.; Binda, C.; Fraaije, M.; Mattevi, A.
    Structure-based enzyme tailoring of 5-hydroxymethylfurfural oxidase (2015), ACS Catal., 5, 1833-1839.
No PubMed abstract available

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.3.47 expressed in Escherichia coli BL21(DE3) cells Methylovorus sp. MP688

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
1.1.3.47 hanging drop vapor diffusion method, using 16-22% (w/v) PEG3350 and 200 mM magnesium formate Methylovorus sp. MP688

Protein Variants

EC Number Protein Variants Comment Organism
1.1.3.47 F434A the mutant shows reduced activity with 4-hydroxy-3-methoxybenzyl alcohol compared to the wild type enzyme Methylovorus sp. MP688
1.1.3.47 H467A the mutant shows reduced activity with 4-hydroxy-3-methoxybenzyl alcohol compared to the wild type enzyme Methylovorus sp. MP688
1.1.3.47 M103A the mutant shows reduced activity with 4-hydroxy-3-methoxybenzyl alcohol compared to the wild type enzyme Methylovorus sp. MP688
1.1.3.47 N511A the mutant shows reduced activity with 4-hydroxy-3-methoxybenzyl alcohol compared to the wild type enzyme Methylovorus sp. MP688
1.1.3.47 V367 the mutant shows increased activity with 5-formylfuran-2-carboxylate compared to the wild type enzyme Methylovorus sp. MP688
1.1.3.47 V367K the mutant shows reduced activity with 4-hydroxy-3-methoxybenzyl alcohol compared to the wild type enzyme Methylovorus sp. MP688
1.1.3.47 V367R/W466F the mutant shows strongly increased activity with 5-formylfuran-2-carboxylate compared to the wild type enzyme Methylovorus sp. MP688
1.1.3.47 V465A the mutant shows reduced activity with 4-hydroxy-3-methoxybenzyl alcohol compared to the wild type enzyme Methylovorus sp. MP688
1.1.3.47 W369A the mutant shows reduced activity with 4-hydroxy-3-methoxybenzyl alcohol compared to the wild type enzyme Methylovorus sp. MP688
1.1.3.47 W466A the mutant shows reduced activity with 4-hydroxy-3-methoxybenzyl alcohol compared to the wild type enzyme Methylovorus sp. MP688
1.1.3.47 W466F the mutant shows increased activity with 5-formylfuran-2-carboxylate compared to the wild type enzyme Methylovorus sp. MP688

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.3.47 0.2
-
 4-Hydroxy-3-methoxybenzyl alcohol mutant enzyme W466F, at pH 7.5 and 25°C Methylovorus sp. MP688
1.1.3.47 0.21
-
 5-formylfuran-2-carboxylate mutant enzyme V367R/W466F, at pH 7.5 and 25°C Methylovorus sp. MP688
1.1.3.47 0.28
-
 4-Hydroxy-3-methoxybenzyl alcohol mutant enzyme V367R, at pH 7.5 and 25°C Methylovorus sp. MP688
1.1.3.47 0.29
-
 4-Hydroxy-3-methoxybenzyl alcohol mutant enzyme V367K, at pH 7.5 and 25°C Methylovorus sp. MP688
1.1.3.47 0.72
-
 4-Hydroxy-3-methoxybenzyl alcohol wild type enzyme, at pH 7.5 and 25°C Methylovorus sp. MP688
1.1.3.47 0.82
-
 4-Hydroxy-3-methoxybenzyl alcohol mutant enzyme H467A, at pH 7.5 and 25°C Methylovorus sp. MP688
1.1.3.47 0.92
-
 4-Hydroxy-3-methoxybenzyl alcohol mutant enzyme V465A, at pH 7.5 and 25°C Methylovorus sp. MP688
1.1.3.47 1
-
 4-Hydroxy-3-methoxybenzyl alcohol mutant enzyme W466A, at pH 7.5 and 25°C Methylovorus sp. MP688
1.1.3.47 1.1
-
 4-Hydroxy-3-methoxybenzyl alcohol mutant enzyme W369A, at pH 7.5 and 25°C Methylovorus sp. MP688
1.1.3.47 1.2
-
 4-Hydroxy-3-methoxybenzyl alcohol mutant enzyme F434A, at pH 7.5 and 25°C Methylovorus sp. MP688
1.1.3.47 1.4
-
 5-formylfuran-2-carboxylate mutant enzyme V367R, at pH 7.5 and 25°C Methylovorus sp. MP688
1.1.3.47 1.8
-
 4-Hydroxy-3-methoxybenzyl alcohol mutant enzyme M103A, at pH 7.5 and 25°C Methylovorus sp. MP688
1.1.3.47 2
-
 4-Hydroxy-3-methoxybenzyl alcohol mutant enzyme N511A, at pH 7.5 and 25°C Methylovorus sp. MP688
1.1.3.47 3
-
 5-formylfuran-2-carboxylate mutant enzyme W466F, at pH 7.5 and 25°C Methylovorus sp. MP688
1.1.3.47 4
-
 5-formylfuran-2-carboxylate Km above 4.0 mM, wild type enzyme, at pH 7.5 and 25°C Methylovorus sp. MP688
1.1.3.47 20
-
 (S)-1-phenylethanol mutant enzyme W466A, at pH 7.5 and 25°C Methylovorus sp. MP688
1.1.3.47 98
-
 (S)-1-phenylethanol mutant enzyme W466F, at pH 7.5 and 25°C Methylovorus sp. MP688

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.1.3.47 5-(dihydroxymethyl)furan-2-carbaldehyde + O2 Methylovorus sp. MP688
-
 5-formylfuran-2-carboxylate + H2O2
-
 ?
1.1.3.47 5-(dihydroxymethyl)furan-2-carboxylate + O2 Methylovorus sp. MP688
-
 furan-2,5-dicarboxylate + H2O2
-
 ?
1.1.3.47 5-(hydroxymethyl)furfural + 3 O2 + 2 H2O Methylovorus sp. MP688 overall reaction furan-2,5-dicarboxylate + 3 H2O2
-
 ?
1.1.3.47 5-(hydroxymethyl)furfural + O2 Methylovorus sp. MP688
-
 furan-2,5-dicarbaldehyde + H2O2
-
 ?
1.1.3.47 5-formylfuran-2-carboxylate + H2O Methylovorus sp. MP688
-
 5-(dihydroxymethyl)furan-2-carboxylate
-
 ?
1.1.3.47 furan-2,5-dicarbaldehyde + H2O Methylovorus sp. MP688
-
 5-(dihydroxymethyl)furan-2-carbaldehyde
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.1.3.47 Methylovorus sp. MP688 E4QP00
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.3.47 HisTrap column chromatography and Superdex 200 gel filtration Methylovorus sp. MP688

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.3.47 (R)-1-phenylethanol + O2
-
 Methylovorus sp. MP688 acetophenone + H2O2
-
 ?
1.1.3.47 (S)-1-phenylethanol + O2
-
 Methylovorus sp. MP688 acetophenone + H2O2
-
 ?
1.1.3.47 4-hydroxy-3-methoxybenzyl alcohol + O2
-
 Methylovorus sp. MP688 4-hydroxy-3-methoxybenzaldehyde + H2O2
-
 ?
1.1.3.47 5-(dihydroxymethyl)furan-2-carbaldehyde + O2
-
 Methylovorus sp. MP688 5-formylfuran-2-carboxylate + H2O2
-
 ?
1.1.3.47 5-(dihydroxymethyl)furan-2-carboxylate + O2
-
 Methylovorus sp. MP688 furan-2,5-dicarboxylate + H2O2
-
 ?
1.1.3.47 5-(hydroxymethyl)furfural + 3 O2 + 2 H2O overall reaction Methylovorus sp. MP688 furan-2,5-dicarboxylate + 3 H2O2
-
 ?
1.1.3.47 5-(hydroxymethyl)furfural + O2
-
 Methylovorus sp. MP688 furan-2,5-dicarbaldehyde + H2O2
-
 ?
1.1.3.47 5-formylfuran-2-carboxylate + H2O
-
 Methylovorus sp. MP688 5-(dihydroxymethyl)furan-2-carboxylate
-
 ?
1.1.3.47 furan-2,5-dicarbaldehyde + H2O
-
 Methylovorus sp. MP688 5-(dihydroxymethyl)furan-2-carbaldehyde
-
 ?

Synonyms

EC Number Synonyms Comment Organism
1.1.3.47 5-hydroxymethylfurfural oxidase
-
 Methylovorus sp. MP688
1.1.3.47 HMFO
-
 Methylovorus sp. MP688

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1.3.47 0.0047
-
 4-Hydroxy-3-methoxybenzyl alcohol mutant enzyme H467A, at pH 7.5 and 25°C Methylovorus sp. MP688
1.1.3.47 0.01
-
 (S)-1-phenylethanol mutant enzyme W466F, at pH 7.5 and 25°C Methylovorus sp. MP688
1.1.3.47 0.011
-
 (S)-1-phenylethanol mutant enzyme W466A, at pH 7.5 and 25°C Methylovorus sp. MP688
1.1.3.47 0.056
-
 5-formylfuran-2-carboxylate mutant enzyme V367R, at pH 7.5 and 25°C Methylovorus sp. MP688
1.1.3.47 0.079
-
 5-formylfuran-2-carboxylate mutant enzyme W466F, at pH 7.5 and 25°C Methylovorus sp. MP688
1.1.3.47 0.46
-
 5-formylfuran-2-carboxylate mutant enzyme V367R/W466F, at pH 7.5 and 25°C Methylovorus sp. MP688
1.1.3.47 0.75
-
 4-Hydroxy-3-methoxybenzyl alcohol mutant enzyme W466A, at pH 7.5 and 25°C Methylovorus sp. MP688
1.1.3.47 1
-
 4-Hydroxy-3-methoxybenzyl alcohol mutant enzyme N511A, at pH 7.5 and 25°C Methylovorus sp. MP688
1.1.3.47 1.5
-
 4-Hydroxy-3-methoxybenzyl alcohol mutant enzyme V465A, at pH 7.5 and 25°C Methylovorus sp. MP688
1.1.3.47 1.7
-
 4-Hydroxy-3-methoxybenzyl alcohol mutant enzyme W466F, at pH 7.5 and 25°C Methylovorus sp. MP688
1.1.3.47 3.3
-
 4-Hydroxy-3-methoxybenzyl alcohol mutant enzyme M103A, at pH 7.5 and 25°C Methylovorus sp. MP688
1.1.3.47 4.3
-
 4-Hydroxy-3-methoxybenzyl alcohol mutant enzyme W369A, at pH 7.5 and 25°C Methylovorus sp. MP688
1.1.3.47 5.1
-
 4-Hydroxy-3-methoxybenzyl alcohol mutant enzyme V367K, at pH 7.5 and 25°C Methylovorus sp. MP688
1.1.3.47 6
-
 4-Hydroxy-3-methoxybenzyl alcohol mutant enzyme V367R, at pH 7.5 and 25°C Methylovorus sp. MP688
1.1.3.47 15
-
 4-Hydroxy-3-methoxybenzyl alcohol mutant enzyme F434A, at pH 7.5 and 25°C Methylovorus sp. MP688
1.1.3.47 21
-
 4-Hydroxy-3-methoxybenzyl alcohol wild type enzyme, at pH 7.5 and 25°C Methylovorus sp. MP688

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.3.47 FAD
-
 Methylovorus sp. MP688

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.1.3.47 0.0001
-
 (S)-1-phenylethanol mutant enzyme W466F, at pH 7.5 and 25°C Methylovorus sp. MP688
1.1.3.47 0.00055
-
 (S)-1-phenylethanol mutant enzyme W466A, at pH 7.5 and 25°C Methylovorus sp. MP688
1.1.3.47 0.005
-
 5-formylfuran-2-carboxylate wild type enzyme, at pH 7.5 and 25°C Methylovorus sp. MP688
1.1.3.47 0.0057
-
 4-Hydroxy-3-methoxybenzyl alcohol mutant enzyme H467A, at pH 7.5 and 25°C Methylovorus sp. MP688
1.1.3.47 0.026
-
 5-formylfuran-2-carboxylate mutant enzyme W466F, at pH 7.5 and 25°C Methylovorus sp. MP688
1.1.3.47 0.041
-
 5-formylfuran-2-carboxylate mutant enzyme V367R, at pH 7.5 and 25°C Methylovorus sp. MP688
1.1.3.47 0.55
-
 4-Hydroxy-3-methoxybenzyl alcohol mutant enzyme N511A, at pH 7.5 and 25°C Methylovorus sp. MP688
1.1.3.47 0.75
-
 4-Hydroxy-3-methoxybenzyl alcohol mutant enzyme W466A, at pH 7.5 and 25°C Methylovorus sp. MP688
1.1.3.47 1.6
-
 4-Hydroxy-3-methoxybenzyl alcohol mutant enzyme V465A, at pH 7.5 and 25°C Methylovorus sp. MP688
1.1.3.47 1.8
-
 4-Hydroxy-3-methoxybenzyl alcohol mutant enzyme M103A, at pH 7.5 and 25°C Methylovorus sp. MP688
1.1.3.47 2.2
-
 5-formylfuran-2-carboxylate mutant enzyme V367R/W466F, at pH 7.5 and 25°C Methylovorus sp. MP688
1.1.3.47 3.9
-
 4-Hydroxy-3-methoxybenzyl alcohol mutant enzyme W369A, at pH 7.5 and 25°C Methylovorus sp. MP688
1.1.3.47 8.5
-
 4-Hydroxy-3-methoxybenzyl alcohol mutant enzyme W466F, at pH 7.5 and 25°C Methylovorus sp. MP688
1.1.3.47 12
-
 4-Hydroxy-3-methoxybenzyl alcohol mutant enzyme F434A, at pH 7.5 and 25°C Methylovorus sp. MP688
1.1.3.47 18
-
 4-Hydroxy-3-methoxybenzyl alcohol mutant enzyme V367K, at pH 7.5 and 25°C Methylovorus sp. MP688
1.1.3.47 21
-
 4-Hydroxy-3-methoxybenzyl alcohol mutant enzyme V367R, at pH 7.5 and 25°C Methylovorus sp. MP688
1.1.3.47 29
-
 4-Hydroxy-3-methoxybenzyl alcohol wild type enzyme, at pH 7.5 and 25°C Methylovorus sp. MP688