Cloned (Comment) | Organism |
---|---|
recombinant expression of His-tagged soluble chloroplastic isoform CuZn-SOD in its enzymatically active and stable form in Escherichia coli, and optimization of culture conditions, best at 18°C and upon isopropyl beta-D-1-thiogalactopyranoside induction, recombinant enzyme production process optimization | Pisum sativum |
Protein Variants | Comment | Organism |
---|---|---|
additional information | the recombinant pea chloroplastic SOD possesses nearly 6fold higher superoxide dismutase activity and 5fold higher peroxidase activity as compared to commercially available CuZn-superoxide dismutase from Bos taurus. The recombinant protein harbors all the characteristics features of this class of enzyme. The recombinant enzyme is exceptionally stable concerning pH and temperature and maintains its activity upon prolonged storage | Pisum sativum |
General Stability | Organism |
---|---|
the purified recombinant enzyme is stable to proteolytic digestion | Pisum sativum |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
diethyldithiocarbamate | complete inhibition at 1.4 mM, a specific inhibitor for CuZn-SODs | Pisum sativum | |
H2O2 | addition of 5 mM of hydrogen peroxide (H2O2) completely inhibits of the enzyme | Pisum sativum | |
KCN | complete inhibition at 0.16 mM, a specific inhibitor for CuZn-SODs | Pisum sativum | |
NaN3 | low inhibition by sodium azide, 40% inhibition at 50 mM | Pisum sativum |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
chloroplast | - |
Pisum sativum | 9507 | - |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Cu2+ | a CuZn-superoxide dismutase, the prosthetic copper and zinc are essential component of CuZn-SODs, 1.80-1.84 Cu2+ per enzyme dimer | Pisum sativum | |
additional information | measurement of Cu and Zn content of PschSOD by ICP-MS, zincon, and bathocuproeine protocols | Pisum sativum | |
Zn2+ | a CuZn-superoxide dismutase, the prosthetic copper and zinc are essential component of CuZn-SODs, 1.6 Zn2+ per enzyme dimer | Pisum sativum |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
36000 | 38000 | recombinant His-tagged enzyme, gel filtration | Pisum sativum |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
2 superoxide + 2 H+ | Pisum sativum | - |
O2 + H2O2 | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pisum sativum | P11964 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged soluble chloroplastic isoform CuZn-SOD from Escherichia coli by nickel affinity chromatography | Pisum sativum |
Storage Stability | Organism |
---|---|
25°C, the purified recombinant enzyme retains 50% of its original activity after storing for 180 days at room temperature | Pisum sativum |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2 superoxide + 2 H+ | - |
Pisum sativum | O2 + H2O2 | - |
? | |
additional information | enzyme PschSOD exhibits superoxide dismutase and peroxidase activities. The enzyme utilizes its own dismutation product, the H2O2 in presence of bicarbonate | Pisum sativum | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
homodimer | 2 * 18000, recombinant His-tagged enzyme, SDS-PAGE | Pisum sativum |
More | circular dichroism spectral analysis of the enzyme's secondary structure. The enzyme conserves the beta-barrel structure of the CuZn-SODs in its recombinant form | Pisum sativum |
Synonyms | Comment | Organism |
---|---|---|
CuZn-SOD | - |
Pisum sativum |
CuZn-superoxide dismutase | - |
Pisum sativum |
PschSOD | - |
Pisum sativum |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Pisum sativum |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
45 | - |
purified recombinant His-tagged enzyme, completely stable at, 60 min | Pisum sativum |
62 | - |
t1/2, purified recombinant His-tagged enzyme, 20-60 min | Pisum sativum |
75 | - |
purified recombinant His-tagged enzyme, 20 min, inactivation | Pisum sativum |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6 | 8 | recombinant His-tagged enzyme | Pisum sativum |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
additional information | - |
the decrease of the enzyme activity at acidic pH might be due to the dissociation of dimer into monomer, as the acidic pH favors the monomer formation, whereas the alkaline pH favors the dimer formation. Charge interaction between the subunits are postulated to be a major determinant of CuZn-SOD activity at varying pHs | Pisum sativum |
5 | 10 | about 70% of maximal activity within this range, 10% at pH 4.0, 40% at pH 11.0 | Pisum sativum |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
additional information | - |
the purified recombinant His-tagged enzyme's activity shows a broad range of pH sustainability | Pisum sativum |
General Information | Comment | Organism |
---|---|---|
additional information | circular dichroism spectral analysis of the enzyme's secondary structure | Pisum sativum |