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Literature summary extracted from

  • Li, M.; Muller, T.A.; Fraser, B.A.; Hausinger, R.P.
    Characterization of active site variants of xanthine hydroxylase from Aspergillus nidulans (2008), Arch. Biochem. Biophys., 470, 44-53.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.14.11.48 wild-type and mutant enzymes Aspergillus nidulans

Protein Variants

EC Number Protein Variants Comment Organism
1.14.11.48 C357A enhanced activity towards 9-methylxanthine as compared to wild-type enzyme. Mutant enzyme is resistant rto thiol-specific reagents that inactivated wild-type enzyme. Elevated ferroxidase activity in the absence of substrates Aspergillus nidulans
1.14.11.48 D138A enhanced activity towards 9-methylxanthine as compared to wild-type enzyme. Significant decrease in Ki(app) for 6,8-dihydroxypurine Aspergillus nidulans
1.14.11.48 D151A inactive mutant enzyme Aspergillus nidulans
1.14.11.48 E137A enhanced activity towards 9-methylxanthine as compared to wild-type enzyme. Significant decrease in Ki(app) for 6,8-dihydroxypurine Aspergillus nidulans
1.14.11.48 H149A inactive mutant enzyme Aspergillus nidulans
1.14.11.48 H340A mutant enzyme exhibits 0.17% of the wild-type enzyme activity Aspergillus nidulans
1.14.11.48 K122A 2fold increase in the Kd of 2-oxoglutarate compared to wild-type enzyme Aspergillus nidulans
1.14.11.48 N358A 23fold decrease in kcat/Km compared to wild-type enzyme. Significant decrease in Ki(app) for 6,8-dihydroxypurine Aspergillus nidulans
1.14.11.48 Q101A elevated ferroxidase activity in the absence of substrates Aspergillus nidulans
1.14.11.48 Q356A significant decrease in Ki(app) for 6,8-dihydroxypurine. Elevated ferroxidase activity in the absence of substrates Aspergillus nidulans

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.14.11.48 6,8-dihydroxypurine slow-binding competitive inhibitor Aspergillus nidulans

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.14.11.48 0.031
-
 2-oxoglutarate pH 7.4, 25°C, wild-type enzyme Aspergillus nidulans
1.14.11.48 0.032
-
 2-oxoglutarate pH 7.4, 25°C, mutant enzyme K122A Aspergillus nidulans
1.14.11.48 0.041
-
 2-oxoglutarate pH 7.4, 25°C, mutant enzyme E137A Aspergillus nidulans
1.14.11.48 0.042
-
 2-oxoglutarate pH 7.4, 25°C, mutant enzyme Q356A Aspergillus nidulans
1.14.11.48 0.045
-
 xanthine pH 7.4, 25°C, wild-type enzyme Aspergillus nidulans
1.14.11.48 0.047
-
 2-oxoglutarate pH 7.4, 25°C, mutant enzyme C357A Aspergillus nidulans
1.14.11.48 0.048
-
 2-oxoglutarate pH 7.4, 25°C, mutant enzyme N358A Aspergillus nidulans
1.14.11.48 0.053
-
 2-oxoglutarate pH 7.4, 25°C, mutant enzyme D138A Aspergillus nidulans
1.14.11.48 0.057
-
 xanthine pH 7.4, 25°C, mutant enzyme Q356A Aspergillus nidulans
1.14.11.48 0.058
-
 xanthine pH 7.4, 25°C, mutant enzyme C357A Aspergillus nidulans
1.14.11.48 0.059
-
 2-oxoglutarate pH 7.4, 25°C, mutant enzyme Q101A Aspergillus nidulans
1.14.11.48 0.069
-
 xanthine pH 7.4, 25°C, mutant enzyme D138A Aspergillus nidulans
1.14.11.48 0.071
-
 xanthine pH 7.4, 25°C, mutant enzyme E137A Aspergillus nidulans
1.14.11.48 0.088
-
 xanthine pH 7.4, 25°C, mutant enzyme K122A Aspergillus nidulans
1.14.11.48 0.122
-
 xanthine pH 7.4, 25°C, mutant enzyme Q101A Aspergillus nidulans
1.14.11.48 0.4
-
 9-methylxanthine pH 7.4, 25°C, wild-type enzyme Aspergillus nidulans
1.14.11.48 0.55
-
 9-methylxanthine pH 7.4, 25°C, mutant enzyme C357A Aspergillus nidulans
1.14.11.48 0.554
-
 xanthine pH 7.4, 25°C, mutant enzyme N358A Aspergillus nidulans
1.14.11.48 0.58
-
 9-methylxanthine pH 7.4, 25°C, mutant enzyme D138A Aspergillus nidulans
1.14.11.48 0.68
-
 9-methylxanthine pH 7.4, 25°C, mutant enzyme Q356A Aspergillus nidulans
1.14.11.48 0.78
-
 9-methylxanthine pH 7.4, 25°C, mutant enzyme Q101A Aspergillus nidulans
1.14.11.48 0.88
-
 9-methylxanthine pH 7.4, 25°C, mutant enzyme K122A Aspergillus nidulans
1.14.11.48 1
-
 9-methylxanthine pH 7.4, 25°C, mutant enzyme N358A Aspergillus nidulans
1.14.11.48 1.09
-
 9-methylxanthine pH 7.4, 25°C, mutant enzyme E137A Aspergillus nidulans

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.14.11.48 Fe2+ non-heme mononuclear Fe(II) enzyme Aspergillus nidulans

Organism

EC Number Organism UniProt Comment Textmining
1.14.11.48 Aspergillus nidulans Q4QZZ9
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.14.11.48 wild-type enzyme variants are purified from recombinant Escherichia coli cells Aspergillus nidulans

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.14.11.48 9-methylxanthine + 2-oxoglutarate + O2
-
 Aspergillus nidulans ?
-
 ?
1.14.11.48 xanthine + 2-oxoglutarate + O2
-
 Aspergillus nidulans urate + succinate + CO2
-
 ?

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.14.11.48 25
-
 assay at Aspergillus nidulans

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.14.11.48 0.66
-
 9-methylxanthine pH 7.4, 25°C, mutant enzyme Q101A Aspergillus nidulans
1.14.11.48 1.26
-
 9-methylxanthine pH 7.4, 25°C, mutant enzyme N358A Aspergillus nidulans
1.14.11.48 1.4
-
 9-methylxanthine pH 7.4, 25°C, mutant enzyme Q356A Aspergillus nidulans
1.14.11.48 1.9
-
 9-methylxanthine pH 7.4, 25°C, mutant enzyme K122A Aspergillus nidulans
1.14.11.48 2 8 2-oxoglutarate pH 7.4, 25°C, mutant enzyme Q356A Aspergillus nidulans
1.14.11.48 3 6 xanthine pH 7.4, 25°C, mutant enzyme Q101A Aspergillus nidulans
1.14.11.48 3.8
-
 9-methylxanthine pH 7.4, 25°C, wild-type enzyme Aspergillus nidulans
1.14.11.48 4.25
-
 9-methylxanthine pH 7.4, 25°C, mutant enzyme D138A Aspergillus nidulans
1.14.11.48 4.5
-
 9-methylxanthine pH 7.4, 25°C, mutant enzyme C357A Aspergillus nidulans
1.14.11.48 5.9
-
 9-methylxanthine pH 7.4, 25°C, mutant enzyme E137A Aspergillus nidulans
1.14.11.48 17
-
 2-oxoglutarate pH 7.4, 25°C, mutant enzyme N358A Aspergillus nidulans
1.14.11.48 19
-
 2-oxoglutarate pH 7.4, 25°C, mutant enzyme C357A Aspergillus nidulans
1.14.11.48 21
-
 2-oxoglutarate pH 7.4, 25°C, mutant enzyme Q101A Aspergillus nidulans
1.14.11.48 25
-
 2-oxoglutarate pH 7.4, 25°C, mutant enzyme D138A Aspergillus nidulans
1.14.11.48 31
-
 2-oxoglutarate pH 7.4, 25°C, mutant enzyme E137A Aspergillus nidulans
1.14.11.48 32
-
 xanthine pH 7.4, 25°C, mutant enzyme C357A Aspergillus nidulans
1.14.11.48 34
-
 xanthine pH 7.4, 25°C, mutant enzyme D138A Aspergillus nidulans
1.14.11.48 35
-
 xanthine pH 7.4, 25°C, mutant enzyme Q356A Aspergillus nidulans
1.14.11.48 38
-
 xanthine pH 7.4, 25°C, mutant enzyme N358A Aspergillus nidulans
1.14.11.48 40
-
 xanthine pH 7.4, 25°C, mutant enzyme E137A Aspergillus nidulans
1.14.11.48 42
-
 2-oxoglutarate pH 7.4, 25°C, mutant enzyme K122A Aspergillus nidulans
1.14.11.48 60
-
 xanthine pH 7.4, 25°C, mutant enzyme K122A Aspergillus nidulans
1.14.11.48 67
-
 2-oxoglutarate pH 7.4, 25°C, wild-type enzyme Aspergillus nidulans
1.14.11.48 72
-
 xanthine pH 7.4, 25°C, wild-type enzyme Aspergillus nidulans

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.14.11.48 7.4
-
 assay at Aspergillus nidulans

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.14.11.48 70
-
 xanthine pH 7.4, 25°C, mutant enzyme N358A Aspergillus nidulans
1.14.11.48 300
-
 xanthine pH 7.4, 25°C, mutant enzyme Q101A Aspergillus nidulans
1.14.11.48 340
-
 2-oxoglutarate pH 7.4, 25°C, mutant enzyme N358A Aspergillus nidulans
1.14.11.48 360
-
 2-oxoglutarate pH 7.4, 25°C, mutant enzyme Q101A Aspergillus nidulans
1.14.11.48 400
-
 2-oxoglutarate pH 7.4, 25°C, mutant enzyme C357A Aspergillus nidulans
1.14.11.48 470
-
 2-oxoglutarate pH 7.4, 25°C, mutant enzyme D138A Aspergillus nidulans
1.14.11.48 490
-
 xanthine pH 7.4, 25°C, mutant enzyme D138A Aspergillus nidulans
1.14.11.48 550
-
 xanthine pH 7.4, 25°C, mutant enzyme C357A Aspergillus nidulans
1.14.11.48 560
-
 xanthine pH 7.4, 25°C, mutant enzyme E137A Aspergillus nidulans
1.14.11.48 610
-
 xanthine pH 7.4, 25°C, mutant enzyme Q356A Aspergillus nidulans
1.14.11.48 670
-
 2-oxoglutarate pH 7.4, 25°C, mutant enzyme Q356A Aspergillus nidulans
1.14.11.48 680
-
 xanthine pH 7.4, 25°C, mutant enzyme K122A Aspergillus nidulans
1.14.11.48 760
-
 2-oxoglutarate pH 7.4, 25°C, mutant enzyme E137A Aspergillus nidulans
1.14.11.48 850
-
 9-methylxanthine pH 7.4, 25°C, mutant enzyme Q101A Aspergillus nidulans
1.14.11.48 1260
-
 9-methylxanthine pH 7.4, 25°C, mutant enzyme N358A Aspergillus nidulans
1.14.11.48 1310
-
 2-oxoglutarate pH 7.4, 25°C, mutant enzyme K122A Aspergillus nidulans
1.14.11.48 1600
-
 xanthine pH 7.4, 25°C, wild-type enzyme Aspergillus nidulans
1.14.11.48 2050
-
 9-methylxanthine pH 7.4, 25°C, mutant enzyme Q356A Aspergillus nidulans
1.14.11.48 2160
-
 9-methylxanthine pH 7.4, 25°C, mutant enzyme K122A Aspergillus nidulans
1.14.11.48 2160
-
 2-oxoglutarate pH 7.4, 25°C, wild-type enzyme Aspergillus nidulans
1.14.11.48 5410
-
 9-methylxanthine pH 7.4, 25°C, mutant enzyme E137A Aspergillus nidulans
1.14.11.48 7330
-
 9-methylxanthine pH 7.4, 25°C, mutant enzyme D138A Aspergillus nidulans
1.14.11.48 8180
-
 9-methylxanthine pH 7.4, 25°C, mutant enzyme C357A Aspergillus nidulans
1.14.11.48 9500
-
 9-methylxanthine pH 7.4, 25°C, wild-type enzyme Aspergillus nidulans