Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary extracted from

  • Paukner, R.; Staudigl, P.; Choosri, W.; Sygmund, C.; Halada, P.; Haltrich, D.; Leitner, C.
    Galactose oxidase from Fusarium oxysporum - expression in E. coli and P. pastoris and biochemical characterization (2014), PLoS ONE, 9, e100116 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.3.9 gene GAOA, sequence comparison to the Fusarium graminearum enzyne, sucessful recombinant expression of C-terminally His-tagged enzyme in Escherichia coli even without codon optimization or further amino acid substitutions. The C-terminal His-tag added to recombinant GalOx from Fusarium oxysporum does not interfere with the catalytic activity. Sucessful recombinant expression of C-terminally His-tagged full-length enzyme and enzyme with the alpha-factor signal sequence of Saccharomyces cerevisiae in Pichia pastoris Fusarium oxysporum

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.3.9 47
-
 D-galactose pH 7.0, 40°C Fusarium oxysporum

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
1.1.3.9 extracellular
-
 Fusarium oxysporum
-
-

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.1.3.9 Cu2+ the enzyme contains a unique metalloradical complex consisting of a copper atom and a tyrosine residue covalently attached to the sulfur of a cysteine Fusarium oxysporum
1.1.3.9 additional information GalOx is unusual among metalloenzymes in catalyzing a two-electron redox chemistry at a mononuclear metal ion active site Fusarium oxysporum

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.1.3.9 D-galactose + O2 Fusarium oxysporum
-
 D-galacto-hexodialdose + H2O2
-
 ?
1.1.3.9 D-galactose + O2 Fusarium oxysporum G12
-
 D-galacto-hexodialdose + H2O2
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.1.3.9 Fusarium oxysporum V5NQ89
-
-
1.1.3.9 Fusarium oxysporum G12 V5NQ89
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.3.9 recombinant C-terminally His-tagged enzyme 28fold from Escherichia coli, recombinant C-terminally His-tagged full-length enzyme (including the native prepro signal sequence) 6fold from Pichia, and recombinant C-terminally His-tagged enzyme with alpha-factor signal sequence 22fold from Pichia pastoris Fusarium oxysporum

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.1.3.9 61
-
 purified recombinant His-tagged full-length enzyme, expressed from Pichia pastoris using the alpha-factor signal of Saccharomyces cerevisiae, pH 7.0, 40°C Fusarium oxysporum
1.1.3.9 63.2
-
 purified recombinant His-tagged full-length enzyme, expressed from Pichia pastoris, pH 7.0, 40°C Fusarium oxysporum
1.1.3.9 65.4
-
 purified recombinant His-tagged enzyme, expressed from Escherichia coli, pH 7.0, 40°C Fusarium oxysporum

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.3.9 1-methyl-beta-D-galactopyranoside + O2 best substrate Fusarium oxysporum ? + H2O2
-
 ?
1.1.3.9 1-methyl-beta-D-galactopyranoside + O2 best substrate Fusarium oxysporum G12 ? + H2O2
-
 ?
1.1.3.9 D-galactose + O2
-
 Fusarium oxysporum D-galacto-hexodialdose + H2O2
-
 ?
1.1.3.9 D-galactose + O2
-
 Fusarium oxysporum G12 D-galacto-hexodialdose + H2O2
-
 ?
1.1.3.9 additional information GalOx catalyzes the oxidation of primary alcohols (e.g. the hydroxyl group at the C6 position in D-galactose) to aldehydes, accompanied by the reduction of molecular oxygen to hydrogen peroxide, and shows a broad substrate tolerance, yet strict stereospecificity, for various alcohol substrates. Because of the high sensitivity of GalOx to the stereo configuration of the C4 hydroxyl group D-glucose is not a substrate for the enzyme Fusarium oxysporum ?
-
 ?
1.1.3.9 additional information GalOx catalyzes the oxidation of primary alcohols (e.g. the hydroxyl group at the C6 position in D-galactose) to aldehydes, accompanied by the reduction of molecular oxygen to hydrogen peroxide, and shows a broad substrate tolerance, yet strict stereospecificity, for various alcohol substrates. Because of the high sensitivity of GalOx to the stereo configuration of the C4 hydroxyl group D-glucose is not a substrate for the enzyme Fusarium oxysporum G12 ?
-
 ?

Subunits

EC Number Subunits Comment Organism
1.1.3.9 monomer 1 * 72000, recombinant His-tagged enzyme, SDS-PAGE, structure analysis, overview Fusarium oxysporum
1.1.3.9 More trypsin digestion and peptide mapping, mass spectrometric analysis Fusarium oxysporum

Synonyms

EC Number Synonyms Comment Organism
1.1.3.9 galactose 6-oxidase
-
 Fusarium oxysporum
1.1.3.9 GalOx
-
 Fusarium oxysporum

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.3.9 40
-
-
 Fusarium oxysporum

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.1.3.9 additional information
-
 the thermostability of GalOx from Fusarium oxysporum expressed in Escherichia coli is significantly lower than the thermostability of enzymes from other Fusarium strains Fusarium oxysporum
1.1.3.9 30
-
 purified recombinant enzyme, pH 7.0, completely stable for at least 24 h Fusarium oxysporum
1.1.3.9 40
-
 purified recombinant enzyme, pH 7.0, loss of 80% activity after 24 h Fusarium oxysporum
1.1.3.9 50
-
 purified recombinant enzyme, pH 7.0, inactivation within 2 h Fusarium oxysporum

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.3.9 7
-
-
 Fusarium oxysporum

pH Range

EC Number pH Minimum pH Maximum Comment Organism
1.1.3.9 6 9 more than 80% of the maximum activity between pH 6.0 and 9.0, maximum activity occurs at pH 7.0 (phosphate buffer) and 8.0 (Tris buffer), respectively. At pH values below pH 5.0 the enzyme loses its activity Fusarium oxysporum

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.3.9 Cys-Tyr cofactor the enzyme contains a unique metalloradical complex consisting of a copper atom and a tyrosine residue covalently attached to the sulfur of a cysteine. The Tyr-Cys crosslink is essential for the catalytic activity of GalOx. The formation of the TyrN-Cys redox cofactor in GalOx is a self-processing reaction requiring only the apoprotein, copper, and dioxygen; no other proteins or enzymes are required for the processing and assembly of the catalytically active enzyme Fusarium oxysporum

General Information

EC Number General Information Comment Organism
1.1.3.9 evolution galactose oxidase is a member of the radical copper oxidase family and is classified as a member of the carbohydrate active-enzyme family AA5, subfamiliy 2 Fusarium oxysporum
1.1.3.9 additional information GalOx is unusual among metalloenzymes in catalyzing a two-electron redox chemistry at a mononuclear metal ion active site Fusarium oxysporum

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.1.3.9 2.2
-
 1-methyl-beta-D-galactopyranoside pH 7.0, 40°C Fusarium oxysporum