Reference to 1.14.11.48; Id = 731289 - BRENDA Enzyme Database

overproduction of His6-tagged enzyme in Aspergillus nidulans and Escherichia coli

Aspergillus nidulans

6,8-dihydroxypurine

0.08 mM, 24% decrease in activity

Aspergillus nidulans

Co2+

0.04 mM, about 65% inhibition

Aspergillus nidulans

Cu2+

0.04 mM, complete inhibition

Aspergillus nidulans

Fe2+

depending on the experimental conditions, Fe2+ concentrations of above 0.025 mM can lead to enzyme inhibition of the recombinant enzyme isolated from Escherichia coli, whereas the enzyme from Aspergillus nidulans is not inhibited at Fe2+ concentrations up to 0.080 mM

Aspergillus nidulans

Mn2+

0.04 mM, about 80% inhibition

Aspergillus nidulans

N-oxalylglycine

competes with 2-oxoglutarate

Aspergillus nidulans

NaCl

0.5 M NaCl increases the Km of 2-oxoglutarate, increases the Km of xanthine and decreased the kcat

Aspergillus nidulans

Zn2+

0.04 mM, complete inhibition

Aspergillus nidulans

0.0311

-

2-oxoglutarate

25Â°C, pH 7.4, recombinant enzyme isolated from Escherichia coli

Aspergillus nidulans

0.0452

-

xanthine

25Â°C, pH 7.4, recombinant enzyme isolated from Escherichia coli

Aspergillus nidulans

0.046

-

xanthine

30Â°C, pH 7.4, enzyme isolated from Aspergillus nidulans

Aspergillus nidulans

0.05

-

2-oxoglutarate

30Â°C, pH 7.4, enzyme isolated from Aspergillus nidulans

Aspergillus nidulans

0.16

-

2-oxoadipate

25Â°C, pH 7.4, recombinant enzyme isolated from Escherichia coli

Aspergillus nidulans

Fe2+

reqired. Half-maximal activity at 0.007 mM when the recombinant apoprotein isolated from the Escherichia coli is used

Aspergillus nidulans

40000

-

12 * 40000, SDS-PAGE

Aspergillus nidulans

500000

-

dodecamer, gel filtration

Aspergillus nidulans

-

glycoprotein

presence of O-glycosylation as well as N-glycosylation

Aspergillus nidulans

purified from both the fungal mycelium and recombinant Escherichia coli cells

Aspergillus nidulans

mycelium

-

Aspergillus nidulans

-

-80Â°C, stable for at least 2 months

Aspergillus nidulans

4Â°C, stored in 100 mM Tris buffer (pH 7) containing 300 mM NaCl, 250 mM imidazole, 15% glycerol, and 0.005 mM Fe2+, concentrated enzyme is stable for at least 5 months

Aspergillus nidulans

4Â°C, stored in 100 mM Tris buffer (pH 8.0) containing 300 mM NaCl, 250 mM imidazole, 1 mM EDTA, and 15% glycerol, stable for at least 1 month

Aspergillus nidulans

additional information

no activity with pyruvate, 2-oxobutyrate, phenyl pyruvate, and 4-hydroxyphenyl pyruvate

731289

Aspergillus nidulans

?

-

?

xanthine + 2-oxoadipate + O2

-

731289

Aspergillus nidulans

?

-

?

xanthine + 2-oxoglutarate + O2

the enzyme is highly specific for xanthine. On the basis of the spectroscopic assay using standard conditions with 12 nM enzyme, no activity is detected when the enzyme is assayed with 0.08, 0.1, or 0.2 mM hypoxanthine, 1-methylxanthine, 3-methylxanthine, 7-methylxanthine, 9-methylxanthine, purine, 6-methylpurine, 2-hydroxypurine, 8-hydroxypurine, 2,8-dihydroxypurine, 2-hydroxy-6-methylpurine, allopurinol, allantoin, or adenosine diphosphate

731289

Aspergillus nidulans

urate + succinate + CO2

-

?

dodecamer

12 * 40000, SDS-PAGE

Aspergillus nidulans

XanA

-

Aspergillus nidulans

xanthine/alpha-ketoglutarate dioxygenase

-

Aspergillus nidulans

7.6

-

2-oxoadipate

25Â°C, pH 7.4, recombinant enzyme isolated from Escherichia coli

Aspergillus nidulans

15

30

2-oxoglutarate

30Â°C, pH 7.4, value depending on preparation, enzyme isolated from Aspergillus nidulans

Aspergillus nidulans

15

30

xanthine

30Â°C, pH 7.4, value depending on preparation, enzyme isolated from Aspergillus nidulans

Aspergillus nidulans

66.5

-

2-oxoglutarate

25Â°C, pH 7.4, recombinant enzyme isolated from Escherichia coli

Aspergillus nidulans

71.4

-

xanthine

25Â°C, pH 7.4, recombinant enzyme isolated from Escherichia coli

Aspergillus nidulans

7

8

the recombinant enzyme isolated from Escherichia coli exhibits a narrow pH optimum of 7.0-8.0 with pH 7.4 yielding optimal activity for Tris, MOPS, MES, imidazole, and HEPES buffers

Aspergillus nidulans

7

-

enzyme isolated from Aspergillus nidulans

Aspergillus nidulans

7

11

stable

Aspergillus nidulans

0.00012

-

N-oxalylglycine

pH 7.4, 25Â°C

Aspergillus nidulans

overproduction of His6-tagged enzyme in Aspergillus nidulans and Escherichia coli

Aspergillus nidulans

6,8-dihydroxypurine

0.08 mM, 24% decrease in activity

Aspergillus nidulans

Co2+

0.04 mM, about 65% inhibition

Aspergillus nidulans

Cu2+

0.04 mM, complete inhibition

Aspergillus nidulans

Fe2+

depending on the experimental conditions, Fe2+ concentrations of above 0.025 mM can lead to enzyme inhibition of the recombinant enzyme isolated from Escherichia coli, whereas the enzyme from Aspergillus nidulans is not inhibited at Fe2+ concentrations up to 0.080 mM

Aspergillus nidulans

Mn2+

0.04 mM, about 80% inhibition

Aspergillus nidulans

N-oxalylglycine

competes with 2-oxoglutarate

Aspergillus nidulans

NaCl

0.5 M NaCl increases the Km of 2-oxoglutarate, increases the Km of xanthine and decreased the kcat

Aspergillus nidulans

Zn2+

0.04 mM, complete inhibition

Aspergillus nidulans

0.00012

-

N-oxalylglycine

pH 7.4, 25Â°C

Aspergillus nidulans

0.0311

-

2-oxoglutarate

25Â°C, pH 7.4, recombinant enzyme isolated from Escherichia coli

Aspergillus nidulans

0.0452

-

xanthine

25Â°C, pH 7.4, recombinant enzyme isolated from Escherichia coli

Aspergillus nidulans

0.046

-

xanthine

30Â°C, pH 7.4, enzyme isolated from Aspergillus nidulans

Aspergillus nidulans

0.05

-

2-oxoglutarate

30Â°C, pH 7.4, enzyme isolated from Aspergillus nidulans

Aspergillus nidulans

0.16

-

2-oxoadipate

25Â°C, pH 7.4, recombinant enzyme isolated from Escherichia coli

Aspergillus nidulans

Fe2+

reqired. Half-maximal activity at 0.007 mM when the recombinant apoprotein isolated from the Escherichia coli is used

Aspergillus nidulans

40000

-

12 * 40000, SDS-PAGE

Aspergillus nidulans

500000

-

dodecamer, gel filtration

Aspergillus nidulans

glycoprotein

presence of O-glycosylation as well as N-glycosylation

Aspergillus nidulans

purified from both the fungal mycelium and recombinant Escherichia coli cells

Aspergillus nidulans

mycelium

-

Aspergillus nidulans

-

-80Â°C, stable for at least 2 months

Aspergillus nidulans

4Â°C, stored in 100 mM Tris buffer (pH 7) containing 300 mM NaCl, 250 mM imidazole, 15% glycerol, and 0.005 mM Fe2+, concentrated enzyme is stable for at least 5 months

Aspergillus nidulans

4Â°C, stored in 100 mM Tris buffer (pH 8.0) containing 300 mM NaCl, 250 mM imidazole, 1 mM EDTA, and 15% glycerol, stable for at least 1 month

Aspergillus nidulans

additional information

no activity with pyruvate, 2-oxobutyrate, phenyl pyruvate, and 4-hydroxyphenyl pyruvate

731289

Aspergillus nidulans

?

-

?

xanthine + 2-oxoadipate + O2

-

731289

Aspergillus nidulans

?

-

?

xanthine + 2-oxoglutarate + O2

the enzyme is highly specific for xanthine. On the basis of the spectroscopic assay using standard conditions with 12 nM enzyme, no activity is detected when the enzyme is assayed with 0.08, 0.1, or 0.2 mM hypoxanthine, 1-methylxanthine, 3-methylxanthine, 7-methylxanthine, 9-methylxanthine, purine, 6-methylpurine, 2-hydroxypurine, 8-hydroxypurine, 2,8-dihydroxypurine, 2-hydroxy-6-methylpurine, allopurinol, allantoin, or adenosine diphosphate

731289

Aspergillus nidulans

urate + succinate + CO2

-

?

dodecamer

12 * 40000, SDS-PAGE

Aspergillus nidulans

7.6

-

2-oxoadipate

25Â°C, pH 7.4, recombinant enzyme isolated from Escherichia coli

Aspergillus nidulans

15

30

2-oxoglutarate

30Â°C, pH 7.4, value depending on preparation, enzyme isolated from Aspergillus nidulans

Aspergillus nidulans

15

30

xanthine

30Â°C, pH 7.4, value depending on preparation, enzyme isolated from Aspergillus nidulans

Aspergillus nidulans

66.5

-

2-oxoglutarate

25Â°C, pH 7.4, recombinant enzyme isolated from Escherichia coli

Aspergillus nidulans

71.4

-

xanthine

25Â°C, pH 7.4, recombinant enzyme isolated from Escherichia coli

Aspergillus nidulans

7

8

the recombinant enzyme isolated from Escherichia coli exhibits a narrow pH optimum of 7.0-8.0 with pH 7.4 yielding optimal activity for Tris, MOPS, MES, imidazole, and HEPES buffers

Aspergillus nidulans

7

-

enzyme isolated from Aspergillus nidulans

Aspergillus nidulans

7

11

stable

Aspergillus nidulans

731217

Li

Characterization of active sit ...

Aspergillus nidulans

Arch. Biochem. Biophys.

470

44-53

2008

-

1

-

10

-

1

24

-

1

-

1

-

1

-

2

-

1

-

24

1

-

1

-

10

-

1

-

24

-

1

-

1

-

2

-

1

-

24

1

-

24

731289

Montero-Moran

Purification and characterizat ...

Aspergillus nidulans

Biochemistry

46

5293-5304

2007

-

1

-

8

5

-

1

2

-

4

-

1

-

1

-

3

1

2

-

5

2

-

1

-

1

-

1

-

8

1

5

-

1

2

-

1

-

1

-

3

1

-

5

2

-

1

-

732527

Cultrone

Convergent evolution of hydrox ...

Aspergillus nidulans

Mol. Microbiol.

57

276-290

2005

-

1

-

1

2

-

1

-

1

-

1

-

1

-

2

1

-

1

-

1

-

1

-

2

-

1

-

1

-

1

-

1

-

1

-

Purification and characterization of the FeII- and alpha-ketoglutarate-dependent xanthine hydroxylase from Aspergillus nidulans

Data extracted from this reference: