BRENDA - Enzyme Database

Purification of a catalase-peroxidase from Halobacterium halobium: Characterization of some unique properties of the halophilic enzyme

Brown-Petersen, N.J.; Salin, M.L.; J. Bacteriol. 175, 4197-4202 (1993)

Data extracted from this reference:

Activating Compound
EC Number
Activating Compound
Commentary
Organism
Structure
1.11.1.6
NaCl
highest activity for catalase at 2 M, for peroxidase at 1 M
Halobacterium salinarum
1.11.1.21
NaCl
peroxidase activity is maximal at NaCl concentrations above 1 M, although catalase activity requires 2 M NaCl for optimal function
Halobacterium salinarum
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
1.11.1.6
3-amino-1H-1,2,4-triazole
not inhibitory
Halobacterium salinarum
1.11.1.6
H2O2
50% inhibition for catalase at 4.5 mM, for peroxidase at 0.4 mM
Halobacterium salinarum
1.11.1.6
KCN
effective inhibition
Halobacterium salinarum
1.11.1.6
NaN3
effective inhibition
Halobacterium salinarum
1.11.1.21
H2O2
-
Halobacterium salinarum
1.11.1.21
KCN
catalase and peroxidase activities are effectively inhibited by KCN
Halobacterium salinarum
1.11.1.21
additional information
catalatic activity of the purified H catalase-peroxidase is not inhibited by incubation for 2.5 h with 20 mM 3 amino-1,2,4-triazole
Halobacterium salinarum
1.11.1.21
NaN3
catalase and peroxidase activities are effectively inhibited by NaN3
Halobacterium salinarum
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.11.1.6
0.14
-
H2O2
peroxidase activity
Halobacterium salinarum
1.11.1.6
3.7
-
H2O2
catalase activity
Halobacterium salinarum
1.11.1.21
0.14
-
H2O2
peroxidase activity, in 50 mM potassium phosphate containing 2 M NaCl, pH 7.5, at 25C
Halobacterium salinarum
1.11.1.21
3.7
-
H2O2
catalase activity, in 50 mM potassium phosphate containing 2 M NaCl, pH 6.5, at 25C
Halobacterium salinarum
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
1.11.1.6
60300
-
4 * 60300, SDS-PAGE
Halobacterium salinarum
1.11.1.6
240000
-
gel filtration
Halobacterium salinarum
1.11.1.21
60300
-
4 * 60300, estimated from SDS-PAGE
Halobacterium salinarum
1.11.1.21
240000
-
gel filtration
Halobacterium salinarum
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.11.1.6
H2O2
Halobacterium salinarum
bifunctional catalase-peroxidase
O2 + H2O
-
Halobacterium salinarum
?
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
1.11.1.6
Halobacterium salinarum
-
-
-
1.11.1.21
Halobacterium salinarum
-
-
-
Purification (Commentary)
EC Number
Commentary
Organism
1.11.1.6
to homogeneity, chromatography steps
Halobacterium salinarum
1.11.1.21
Macrosphere GPC 300 column chromatography, hydroxylapatite column chromatography, and Sephadex G-150 gel filtration
Halobacterium salinarum
Source Tissue
EC Number
Source Tissue
Commentary
Organism
Textmining
1.11.1.6
cell culture
-
Halobacterium salinarum
-
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [mol/min/mg]
Specific Activity Maximum [mol/min/mg]
Commentary
Organism
1.11.1.6
43.2
-
catalase activity
Halobacterium salinarum
1.11.1.6
104.3
-
peroxidase activity
Halobacterium salinarum
1.11.1.21
3.4
-
crude extract, catalase activity, in 50 mM potassium phosphate containing 2 M NaCl, pH 6.5, at 25C
Halobacterium salinarum
1.11.1.21
3.9
-
crude extract, peroxidase activity, in 50 mM potassium phosphate containing 2 M NaCl, pH 7.5, at 25C
Halobacterium salinarum
1.11.1.21
43.2
-
after 12.7fold purification, catalase activity, in 50 mM potassium phosphate containing 2 M NaCl, pH 6.5, at 25C
Halobacterium salinarum
1.11.1.21
104.3
-
after 29.6fold purification, peroxidase activity, in 50 mM potassium phosphate containing 2 M NaCl, pH 7.5, at 25C
Halobacterium salinarum
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.11.1.6
3,3'-diaminobenzidine + H2O2
-
439789
Halobacterium salinarum
? + H2O
-
439789
Halobacterium salinarum
?
1.11.1.6
4-aminoantipyrine + H2O2
-
439789
Halobacterium salinarum
? + H2O
-
439789
Halobacterium salinarum
?
1.11.1.6
ascorbate + H2O2
-
439789
Halobacterium salinarum
? + H2O
-
439789
Halobacterium salinarum
?
1.11.1.6
H2O2
bifunctional catalase-peroxidase
439789
Halobacterium salinarum
O2 + H2O
-
439789
Halobacterium salinarum
?
1.11.1.6
NADH + H2O2
-
439789
Halobacterium salinarum
NAD+ + H2O
-
439789
Halobacterium salinarum
?
1.11.1.6
NADPH + H2O2
-
439789
Halobacterium salinarum
NADP+ + H2O
-
439789
Halobacterium salinarum
?
1.11.1.6
o-dianisidine + H2O2
-
439789
Halobacterium salinarum
? + H2O
-
439789
Halobacterium salinarum
?
1.11.1.6
reduced cytochrome c + H2O2
-
439789
Halobacterium salinarum
oxidized cytochrome c + H2O
-
439789
Halobacterium salinarum
?
1.11.1.21
3,3'-diaminobenzidine + H2O2
-
439789
Halobacterium salinarum
oxidized 3,3'-diaminobenzidine + H2O
-
-
-
?
1.11.1.21
4-aminoantipyrine + H2O2
-
439789
Halobacterium salinarum
oxidized 4-aminoantipyrine + H2O
-
-
-
?
1.11.1.21
ascorbate + H2O2
-
439789
Halobacterium salinarum
monodehydroascorbate + H2O
-
-
-
?
1.11.1.21
H2O2
-
439789
Halobacterium salinarum
O2 + H2O
-
-
-
?
1.11.1.21
NADH + H2O2
-
439789
Halobacterium salinarum
?
-
-
-
?
1.11.1.21
NADPH + H2O2
-
439789
Halobacterium salinarum
?
-
-
-
?
1.11.1.21
o-dianisidine + H2O2
-
439789
Halobacterium salinarum
oxidized o-dianisidine + H2O
-
-
-
?
1.11.1.21
reduced cytochrome c + H2O2
-
439789
Halobacterium salinarum
oxidized cytochrome c + H2O
-
-
-
?
Subunits
EC Number
Subunits
Commentary
Organism
1.11.1.6
tetramer
4 * 60300, SDS-PAGE
Halobacterium salinarum
1.11.1.21
homotetramer
4 * 60300, estimated from SDS-PAGE
Halobacterium salinarum
Temperature Optimum [C]
EC Number
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
1.11.1.6
40
-
peroxidase activity
Halobacterium salinarum
1.11.1.6
50
-
catalase activity
Halobacterium salinarum
1.11.1.21
40
-
peroxidase activity decreases rapidly above its maximum at 40C
Halobacterium salinarum
1.11.1.21
50
-
catalase activity
Halobacterium salinarum
Temperature Stability [C]
EC Number
Temperature Stability Minimum [C]
Temperature Stability Maximum [C]
Commentary
Organism
1.11.1.6
70
-
50% of activity retained for peroxidase activity
Halobacterium salinarum
1.11.1.6
90
-
catalase activity stable up to
Halobacterium salinarum
1.11.1.21
25
90
at 90C, the catalatic activity is 20% greater than at 25C. Peroxidatic activity declines after incubation at temperatures higher than 40C, although at 70C nearly 50% of the activity at 25C is retained
Halobacterium salinarum
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.11.1.6
6.5
-
catalase activity
Halobacterium salinarum
1.11.1.6
7.5
-
peroxidase activity
Halobacterium salinarum
pH Range
EC Number
pH Minimum
pH Maximum
Commentary
Organism
1.11.1.21
6
7.5
catalase activity
Halobacterium salinarum
1.11.1.21
6.5
8
peroxidase activity
Halobacterium salinarum
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
1.11.1.6
heme
1.43 hemes per tetramer
Halobacterium salinarum
1.11.1.21
heme
heme content of 1.43 per tetramer
Halobacterium salinarum
Ki Value [mM]
EC Number
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
1.11.1.6
0.03
-
NaN3
-
Halobacterium salinarum
1.11.1.6
0.05
-
KCN
-
Halobacterium salinarum
1.11.1.21
0.03
-
NaN3
apparent value, peroxidase activity, in 50 mM potassium phosphate containing 2 M NaCl, pH 7.5, at 25C
Halobacterium salinarum
1.11.1.21
0.05
-
KCN
apparent value, catalase activity, in 50 mM potassium phosphate containing 2 M NaCl, pH 6.5, at 25C; apparent value, peroxidase activity, in 50 mM potassium phosphate containing 2 M NaCl, pH 7.5, at 25C
Halobacterium salinarum
1.11.1.21
0.0675
-
NaN3
apparent value, catalase activity, in 50 mM potassium phosphate containing 2 M NaCl, pH 6.5, at 25C
Halobacterium salinarum
pI Value
EC Number
Organism
Commentary
pI Value Maximum
pI Value
1.11.1.21
Halobacterium salinarum
isoelectric focusing
-
3.8
IC50 Value
EC Number
IC50 Value
IC50 Value Maximum
Commentary
Organism
Inhibitor
Structure
1.11.1.21
0.4
-
peroxidase activity, in 50 mM potassium phosphate containing 2 M NaCl, pH 7.5, at 25C
Halobacterium salinarum
H2O2
1.11.1.21
4.5
-
catalase activity, in 50 mM potassium phosphate containing 2 M NaCl, pH 6.5, at 25C
Halobacterium salinarum
H2O2
Activating Compound (protein specific)
EC Number
Activating Compound
Commentary
Organism
Structure
1.11.1.6
NaCl
highest activity for catalase at 2 M, for peroxidase at 1 M
Halobacterium salinarum
1.11.1.21
NaCl
peroxidase activity is maximal at NaCl concentrations above 1 M, although catalase activity requires 2 M NaCl for optimal function
Halobacterium salinarum
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
1.11.1.6
heme
1.43 hemes per tetramer
Halobacterium salinarum
1.11.1.21
heme
heme content of 1.43 per tetramer
Halobacterium salinarum
IC50 Value (protein specific)
EC Number
IC50 Value
IC50 Value Maximum
Commentary
Organism
Inhibitor
Structure
1.11.1.21
0.4
-
peroxidase activity, in 50 mM potassium phosphate containing 2 M NaCl, pH 7.5, at 25C
Halobacterium salinarum
H2O2
1.11.1.21
4.5
-
catalase activity, in 50 mM potassium phosphate containing 2 M NaCl, pH 6.5, at 25C
Halobacterium salinarum
H2O2
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
1.11.1.6
3-amino-1H-1,2,4-triazole
not inhibitory
Halobacterium salinarum
1.11.1.6
H2O2
50% inhibition for catalase at 4.5 mM, for peroxidase at 0.4 mM
Halobacterium salinarum
1.11.1.6
KCN
effective inhibition
Halobacterium salinarum
1.11.1.6
NaN3
effective inhibition
Halobacterium salinarum
1.11.1.21
H2O2
-
Halobacterium salinarum
1.11.1.21
KCN
catalase and peroxidase activities are effectively inhibited by KCN
Halobacterium salinarum
1.11.1.21
additional information
catalatic activity of the purified H catalase-peroxidase is not inhibited by incubation for 2.5 h with 20 mM 3 amino-1,2,4-triazole
Halobacterium salinarum
1.11.1.21
NaN3
catalase and peroxidase activities are effectively inhibited by NaN3
Halobacterium salinarum
Ki Value [mM] (protein specific)
EC Number
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
1.11.1.6
0.03
-
NaN3
-
Halobacterium salinarum
1.11.1.6
0.05
-
KCN
-
Halobacterium salinarum
1.11.1.21
0.03
-
NaN3
apparent value, peroxidase activity, in 50 mM potassium phosphate containing 2 M NaCl, pH 7.5, at 25C
Halobacterium salinarum
1.11.1.21
0.05
-
KCN
apparent value, catalase activity, in 50 mM potassium phosphate containing 2 M NaCl, pH 6.5, at 25C; apparent value, peroxidase activity, in 50 mM potassium phosphate containing 2 M NaCl, pH 7.5, at 25C
Halobacterium salinarum
1.11.1.21
0.0675
-
NaN3
apparent value, catalase activity, in 50 mM potassium phosphate containing 2 M NaCl, pH 6.5, at 25C
Halobacterium salinarum
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.11.1.6
0.14
-
H2O2
peroxidase activity
Halobacterium salinarum
1.11.1.6
3.7
-
H2O2
catalase activity
Halobacterium salinarum
1.11.1.21
0.14
-
H2O2
peroxidase activity, in 50 mM potassium phosphate containing 2 M NaCl, pH 7.5, at 25C
Halobacterium salinarum
1.11.1.21
3.7
-
H2O2
catalase activity, in 50 mM potassium phosphate containing 2 M NaCl, pH 6.5, at 25C
Halobacterium salinarum
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
1.11.1.6
60300
-
4 * 60300, SDS-PAGE
Halobacterium salinarum
1.11.1.6
240000
-
gel filtration
Halobacterium salinarum
1.11.1.21
60300
-
4 * 60300, estimated from SDS-PAGE
Halobacterium salinarum
1.11.1.21
240000
-
gel filtration
Halobacterium salinarum
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.11.1.6
H2O2
Halobacterium salinarum
bifunctional catalase-peroxidase
O2 + H2O
-
Halobacterium salinarum
?
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
1.11.1.6
to homogeneity, chromatography steps
Halobacterium salinarum
1.11.1.21
Macrosphere GPC 300 column chromatography, hydroxylapatite column chromatography, and Sephadex G-150 gel filtration
Halobacterium salinarum
Source Tissue (protein specific)
EC Number
Source Tissue
Commentary
Organism
Textmining
1.11.1.6
cell culture
-
Halobacterium salinarum
-
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [mol/min/mg]
Specific Activity Maximum [mol/min/mg]
Commentary
Organism
1.11.1.6
43.2
-
catalase activity
Halobacterium salinarum
1.11.1.6
104.3
-
peroxidase activity
Halobacterium salinarum
1.11.1.21
3.4
-
crude extract, catalase activity, in 50 mM potassium phosphate containing 2 M NaCl, pH 6.5, at 25C
Halobacterium salinarum
1.11.1.21
3.9
-
crude extract, peroxidase activity, in 50 mM potassium phosphate containing 2 M NaCl, pH 7.5, at 25C
Halobacterium salinarum
1.11.1.21
43.2
-
after 12.7fold purification, catalase activity, in 50 mM potassium phosphate containing 2 M NaCl, pH 6.5, at 25C
Halobacterium salinarum
1.11.1.21
104.3
-
after 29.6fold purification, peroxidase activity, in 50 mM potassium phosphate containing 2 M NaCl, pH 7.5, at 25C
Halobacterium salinarum
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.11.1.6
3,3'-diaminobenzidine + H2O2
-
439789
Halobacterium salinarum
? + H2O
-
439789
Halobacterium salinarum
?
1.11.1.6
4-aminoantipyrine + H2O2
-
439789
Halobacterium salinarum
? + H2O
-
439789
Halobacterium salinarum
?
1.11.1.6
ascorbate + H2O2
-
439789
Halobacterium salinarum
? + H2O
-
439789
Halobacterium salinarum
?
1.11.1.6
H2O2
bifunctional catalase-peroxidase
439789
Halobacterium salinarum
O2 + H2O
-
439789
Halobacterium salinarum
?
1.11.1.6
NADH + H2O2
-
439789
Halobacterium salinarum
NAD+ + H2O
-
439789
Halobacterium salinarum
?
1.11.1.6
NADPH + H2O2
-
439789
Halobacterium salinarum
NADP+ + H2O
-
439789
Halobacterium salinarum
?
1.11.1.6
o-dianisidine + H2O2
-
439789
Halobacterium salinarum
? + H2O
-
439789
Halobacterium salinarum
?
1.11.1.6
reduced cytochrome c + H2O2
-
439789
Halobacterium salinarum
oxidized cytochrome c + H2O
-
439789
Halobacterium salinarum
?
1.11.1.21
3,3'-diaminobenzidine + H2O2
-
439789
Halobacterium salinarum
oxidized 3,3'-diaminobenzidine + H2O
-
-
-
?
1.11.1.21
4-aminoantipyrine + H2O2
-
439789
Halobacterium salinarum
oxidized 4-aminoantipyrine + H2O
-
-
-
?
1.11.1.21
ascorbate + H2O2
-
439789
Halobacterium salinarum
monodehydroascorbate + H2O
-
-
-
?
1.11.1.21
H2O2
-
439789
Halobacterium salinarum
O2 + H2O
-
-
-
?
1.11.1.21
NADH + H2O2
-
439789
Halobacterium salinarum
?
-
-
-
?
1.11.1.21
NADPH + H2O2
-
439789
Halobacterium salinarum
?
-
-
-
?
1.11.1.21
o-dianisidine + H2O2
-
439789
Halobacterium salinarum
oxidized o-dianisidine + H2O
-
-
-
?
1.11.1.21
reduced cytochrome c + H2O2
-
439789
Halobacterium salinarum
oxidized cytochrome c + H2O
-
-
-
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
1.11.1.6
tetramer
4 * 60300, SDS-PAGE
Halobacterium salinarum
1.11.1.21
homotetramer
4 * 60300, estimated from SDS-PAGE
Halobacterium salinarum
Temperature Optimum [C] (protein specific)
EC Number
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
1.11.1.6
40
-
peroxidase activity
Halobacterium salinarum
1.11.1.6
50
-
catalase activity
Halobacterium salinarum
1.11.1.21
40
-
peroxidase activity decreases rapidly above its maximum at 40C
Halobacterium salinarum
1.11.1.21
50
-
catalase activity
Halobacterium salinarum
Temperature Stability [C] (protein specific)
EC Number
Temperature Stability Minimum [C]
Temperature Stability Maximum [C]
Commentary
Organism
1.11.1.6
70
-
50% of activity retained for peroxidase activity
Halobacterium salinarum
1.11.1.6
90
-
catalase activity stable up to
Halobacterium salinarum
1.11.1.21
25
90
at 90C, the catalatic activity is 20% greater than at 25C. Peroxidatic activity declines after incubation at temperatures higher than 40C, although at 70C nearly 50% of the activity at 25C is retained
Halobacterium salinarum
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.11.1.6
6.5
-
catalase activity
Halobacterium salinarum
1.11.1.6
7.5
-
peroxidase activity
Halobacterium salinarum
pH Range (protein specific)
EC Number
pH Minimum
pH Maximum
Commentary
Organism
1.11.1.21
6
7.5
catalase activity
Halobacterium salinarum
1.11.1.21
6.5
8
peroxidase activity
Halobacterium salinarum
pI Value (protein specific)
EC Number
Organism
Commentary
pI Value Maximum
pI Value
1.11.1.21
Halobacterium salinarum
isoelectric focusing
-
3.8