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A novel thermostable sulfite oxidase from Thermus thermophilus: characterization of the enzyme, gene cloning and expression in Escherichia coli

Di Salle, A.; DErrico, G.; La Cara, F.; Cannio, R.; Rossi, M.; Extremophiles 10, 587-598 (2006)

Data extracted from this reference:

Activating Compound
EC Number
Activating Compound
Commentary
Organism
Structure
1.8.3.1
EDTA
-
Thermus thermophilus
1.8.3.1
Molybdenum
the addition of molybdenum to culture media at a concentration of 2.07 mM molybdate leads to a 4fold increase in activity
Thermus thermophilus
1.8.3.1
additional information
molybdenum concentrations higher than 2.07 mM do not show increase of enzyme activity
Thermus thermophilus
Cloned(Commentary)
EC Number
Commentary
Organism
1.8.3.1
expressed in Escherichia coli strain TP1000
Thermus thermophilus
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
1.8.3.1
arsenite
7.4 mM, 50% inhibition
Thermus thermophilus
1.8.3.1
Ca2+
-
Thermus thermophilus
1.8.3.1
additional information
inhibition at 400 mM salt concentrations; no inhibition by periodate or 80 mM sulfate
Thermus thermophilus
Localization
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
1.8.3.1
periplasm
-
Thermus thermophilus
-
-
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
1.8.3.1
K+
45-100 mM, 54-88% increase of activity
Thermus thermophilus
1.8.3.1
Molybdenum
the addition of molybdenum to culture media at a concentration of 2.07 mM molybdate leads to a 4fold increase in activity
Thermus thermophilus
1.8.3.1
NH4+
30 mM, 74% increase of activity
Thermus thermophilus
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
1.8.3.1
38000
-
gel filtration
Thermus thermophilus
1.8.3.1
39100
-
SDS-PAGE
Thermus thermophilus
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
1.8.3.1
Thermus thermophilus
-
-
-
1.8.3.1
Thermus thermophilus AT62
-
-
-
Purification (Commentary)
EC Number
Commentary
Organism
1.8.3.1
DEAE 52 column chromatography, Superdex X26 gel filtration and hydroxyapatite column chromatography
Thermus thermophilus
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
1.8.3.1
0.191
-
crude cell extract
Thermus thermophilus
1.8.3.1
56.67
-
after 297fold purification
Thermus thermophilus
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.8.3.1
sulfite + H2O + A
-
668531
Thermus thermophilus
SO42- + AH2
-
-
-
?
1.8.3.1
sulfite + H2O + A
-
668531
Thermus thermophilus AT62
SO42- + AH2
-
-
-
?
Temperature Optimum [°C]
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
1.8.3.1
60
-
-
Thermus thermophilus
Temperature Stability [°C]
EC Number
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
1.8.3.1
50
80
The enzyme retains 100% of residual activity up to 7 h of incubation at 50°C. Half life times of the enzyme at 60, 70 and 80°C are respectively 8, 6.5 and 1.5 h. The enzyme retains 30% of activity at 25°C and 45% at 90°C.
Thermus thermophilus
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1.8.3.1
0.4
-
sulfite
-
Thermus thermophilus
1.8.3.1
53.32
-
sulfite
-
Thermus thermophilus
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.8.3.1
11
-
-
Thermus thermophilus
Ki Value [mM]
EC Number
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
1.8.3.1
9.4
-
Ca2+
-
Thermus thermophilus
pI Value
EC Number
Organism
Commentary
pI Value Maximum
pI Value
1.8.3.1
Thermus thermophilus
-
-
6.8
Activating Compound (protein specific)
EC Number
Activating Compound
Commentary
Organism
Structure
1.8.3.1
EDTA
-
Thermus thermophilus
1.8.3.1
Molybdenum
the addition of molybdenum to culture media at a concentration of 2.07 mM molybdate leads to a 4fold increase in activity
Thermus thermophilus
1.8.3.1
additional information
molybdenum concentrations higher than 2.07 mM do not show increase of enzyme activity
Thermus thermophilus
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
1.8.3.1
expressed in Escherichia coli strain TP1000
Thermus thermophilus
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
1.8.3.1
arsenite
7.4 mM, 50% inhibition
Thermus thermophilus
1.8.3.1
Ca2+
-
Thermus thermophilus
1.8.3.1
additional information
inhibition at 400 mM salt concentrations; no inhibition by periodate or 80 mM sulfate
Thermus thermophilus
Ki Value [mM] (protein specific)
EC Number
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
1.8.3.1
9.4
-
Ca2+
-
Thermus thermophilus
Localization (protein specific)
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
1.8.3.1
periplasm
-
Thermus thermophilus
-
-
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
1.8.3.1
K+
45-100 mM, 54-88% increase of activity
Thermus thermophilus
1.8.3.1
Molybdenum
the addition of molybdenum to culture media at a concentration of 2.07 mM molybdate leads to a 4fold increase in activity
Thermus thermophilus
1.8.3.1
NH4+
30 mM, 74% increase of activity
Thermus thermophilus
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
1.8.3.1
38000
-
gel filtration
Thermus thermophilus
1.8.3.1
39100
-
SDS-PAGE
Thermus thermophilus
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
1.8.3.1
DEAE 52 column chromatography, Superdex X26 gel filtration and hydroxyapatite column chromatography
Thermus thermophilus
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
1.8.3.1
0.191
-
crude cell extract
Thermus thermophilus
1.8.3.1
56.67
-
after 297fold purification
Thermus thermophilus
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.8.3.1
sulfite + H2O + A
-
668531
Thermus thermophilus
SO42- + AH2
-
-
-
?
1.8.3.1
sulfite + H2O + A
-
668531
Thermus thermophilus AT62
SO42- + AH2
-
-
-
?
Temperature Optimum [°C] (protein specific)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
1.8.3.1
60
-
-
Thermus thermophilus
Temperature Stability [°C] (protein specific)
EC Number
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
1.8.3.1
50
80
The enzyme retains 100% of residual activity up to 7 h of incubation at 50°C. Half life times of the enzyme at 60, 70 and 80°C are respectively 8, 6.5 and 1.5 h. The enzyme retains 30% of activity at 25°C and 45% at 90°C.
Thermus thermophilus
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1.8.3.1
0.4
-
sulfite
-
Thermus thermophilus
1.8.3.1
53.32
-
sulfite
-
Thermus thermophilus
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.8.3.1
11
-
-
Thermus thermophilus
pI Value (protein specific)
EC Number
Organism
Commentary
pI Value Maximum
pI Value
1.8.3.1
Thermus thermophilus
-
-
6.8