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Literature summary for 1.1.3.9 extracted from

  • Rogers, M.S.; Tyler, E.M.; Akyumani, N.; Kurtis, C.R.; Spooner, R.K.; Deacon, S.E.; Tamber, S.; Firbank, S.J.; Mahmoud, K.; Knowles, P.F.; Phillips, S.E.; McPherson, M.J.; Dooley, D.M.
    The stacking tryptophan of galactose oxidase: a second-coordination sphere residue that has profound effects on tyrosyl radical behavior and enzyme catalysis (2007), Biochemistry, 46, 4606-4618.
    View publication on PubMedView publication on EuropePMC

Crystallization (Commentary)

Crystallization (Comment) Organism
crystals of W290F and W290G are grown from protein as isolated, using hanging drop vapor diffusion from 1 to 2 M (NH4)2SO4 and 0.1 M sodium acetate at pH 4.0-5.0. To prepare the azide complex, wild-type galactose oxidase is dialyzed into 50 mM PIPES (pH 7.0) for 2 h, before addition of 20 mM sodium iethyl dithiocarbamate (DDC). The copper chelator DDC promotes the formation of orthorhombic crystals, which have more favorable crystal packing for substrate soaking experiments. The copper-free protein is crystallized in 13.5% PEG 8000, 200 mM calcium acetate, and 100 mM MES (pH 6.3) Fusarium graminearum

Protein Variants

Protein Variants Comment Organism
W290F kcat/KM for D-galactose is 49fold lower than wild-type value, kcat/Km for 2-methylene-1,3-propanediol is 1.2fold higher than wild-type value Fusarium graminearum
W290G kcat/KM for D-galactose is 6370fold lower than wild-type value Fusarium graminearum
W290H kcat/KM for D-galactose is 1180fold lower than wild-type value Fusarium graminearum

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
29
-
2-methylene-1,3-propanediol 25°C, pH 7.0, mutant enzyme W290F Fusarium graminearum
45
-
D-galactose 25°C, pH 7.0, mutant enzyme W290H Fusarium graminearum
57
-
2-methylene-1,3-propanediol 25°C, pH 7.0, wild-type enzyme Fusarium graminearum
82
-
D-galactose 25°C, pH 7.0, wild-type enzyme Fusarium graminearum
1686
-
D-galactose 25°C, pH 7.0, mutant enzyme W290G Fusarium graminearum
2950
-
D-galactose 25°C, pH 7.0, mutant enzyme W290F Fusarium graminearum

Metals/Ions

Metals/Ions Comment Organism Structure
copper copper content: 1.2 mol of Cu per mol of wild-type enzyme, 0.96 mol of Cu per mol of mutant enzyme W290F, 1 mol Cu per mol of mutant enzyme W290G, 1.3 mol of Cu per mol of mutant enzyme W290H Fusarium graminearum

Organism

Organism UniProt Comment Textmining
Fusarium graminearum P0CS93
-
-

Purification (Commentary)

Purification (Comment) Organism
-
Fusarium graminearum

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2-methylene-1,3-propanediol + O2
-
Fusarium graminearum ?
-
?
D-galactose + O2
-
Fusarium graminearum D-galacto-hexodialdose + H2O2
-
?

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.24
-
D-galactose 25°C, pH 7.0, mutant enzyme W290H Fusarium graminearum
1.66
-
D-galactose 25°C, pH 7.0, mutant enzyme W290G Fusarium graminearum
166
-
2-methylene-1,3-propanediol 25°C, pH 7.0, mutant enzyme W290F Fusarium graminearum
283
-
2-methylene-1,3-propanediol 25°C, pH 7.0, wild-type enzyme Fusarium graminearum
371
-
D-galactose 25°C, pH 7.0, mutant enzyme W290F Fusarium graminearum
503
-
D-galactose 25°C, pH 7.0, wild-type enzyme Fusarium graminearum