Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli, the presence of 3% NaCl as well as beta-D-thiogalactopyranoside is needed for the expression | Chlamydomonas sp. |
General Stability | Organism |
---|---|
the enzyme is relatively stable in ascorbate-depleted medium | Chlamydomonas sp. |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
KCN | complete inhibition at 0.1 mM | Chlamydomonas sp. | |
NaN3 | complete inhibition at 4 mM | Chlamydomonas sp. | |
p-chloromercuribenzoate | 84% inhibition at 0.2 mM for 5 min | Chlamydomonas sp. |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.05 | - |
H2O2 | native and recombinant enzyme | Chlamydomonas sp. | |
0.34 | - |
ascorbate | native and recombinant enzyme | Chlamydomonas sp. |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
chloroplast stroma | - |
Chlamydomonas sp. | 9570 | - |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
30030 | - |
calculated from the cDNA clone encoding a mature protein of 282 amino acids | Chlamydomonas sp. |
31000 | - |
gel filtration, native and recombinant enzyme | Chlamydomonas sp. |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Chlamydomonas sp. | Q9SXL5 | W80 strain | - |
Purification (Comment) | Organism |
---|---|
of the native enzyme, using ammonium sulfate precipitation, column chromatography on Phenyl-Sepharose, second ammonium sulfate precipitation, and column chromatography on Superdex 200 and Q Sepharose | Chlamydomonas sp. |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
1.3 | - |
recombinant enzyme, soluble fraction, addition of 3% NaCl | Chlamydomonas sp. |
580 | - |
purified recombinant enzyme | Chlamydomonas sp. |
636 | - |
purified native enzyme | Chlamydomonas sp. |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
cytochrome c + H2O2 | no activity | Chlamydomonas sp. | ? + H2O | - |
? | |
D-iso-ascorbate + H2O2 | native enzyme: the activity with D-isoascorbate corresponds to 131% of that found with ascorbate, recombinant enzyme: the activity with D-isoascorbate corresponds to 129% of that found with ascorbate | Chlamydomonas sp. | dehydroascorbate + H2O | - |
? | |
glutathione + H2O2 | no activity | Chlamydomonas sp. | ? + H2O | - |
? | |
guaiacol + H2O2 | native enzyme: the activity corresponds to 7.2% of that found with L-ascorbate, recombinant enzyme: the activity corresponds to 8% of that found with L-ascorbate | Chlamydomonas sp. | ? | - |
? | |
L-ascorbate + H2O2 | native and recombinant enzyme, no activation is observed, when the enzyme is incubated with H2O2 under anaerobic conditions, thus one of the reasons for the stability mechanism in the enzyme may be the insusceptibility of compound I to H2O2 | Chlamydomonas sp. | dehydroascorbate + H2O | - |
? | |
L-ascorbic acid + cumene hydroperoxide | no activity | Chlamydomonas sp. | dehydroascorbate + 1,1-dimethylbenzylalcohol + H2O | - |
? | |
L-ascorbic acid + tert-butylhydroperoxide | no activity | Chlamydomonas sp. | dehydroascorbate + tert-butylalcohol | - |
? | |
additional information | native and recombinant enzyme, no activity with: glutathione, NADPH and cytochrome c | Chlamydomonas sp. | ? | - |
? | |
NADPH + H2O2 | no activity | Chlamydomonas sp. | ? + H2O | - |
? | |
pyrogallol + H2O2 | native enzyme: the activity corresponds to 121% of that found with L-ascorbate, recombinant enzyme: the activity corresponds to 130% of that found with L-ascorbate | Chlamydomonas sp. | 3-hydroxybenzo-1,2-quinone + H2O | - |
? |
Subunits | Comment | Organism |
---|---|---|
monomer | 1 * 31000, SDS-PAGE, native and recombinant enzyme | Chlamydomonas sp. |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
42 | - |
- |
Chlamydomonas sp. |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6.8 | - |
- |
Chlamydomonas sp. |
7 | - |
assay at | Chlamydomonas sp. |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
heme | - |
Chlamydomonas sp. |