Cloned (Comment) | Organism |
---|---|
expression in Pichia pastoris | Aspergillus niger |
gene phyA, expression in Pichia pastoris strain X33, subcloning in Escherichia coli strain JM109 and DH5alpha | Aspergillus niger |
Protein Variants | Comment | Organism |
---|---|---|
C215S | removal of disulfide bridge results in increase in Km, decrease in kcat value, loss of activity at 58°C | Aspergillus niger |
C215S | site-directed mutagenesis, the mutant shows altered temperature optimum compared to the wild-type enzyme | Aspergillus niger |
C264G | removal of disulfide bridge results in decrease in kcat value, loss of activity at 58°C | Aspergillus niger |
C264G | site-directed mutagenesis, the mutant shows altered pH and temperature optima compared to the wild-type enzyme | Aspergillus niger |
C31G | removal of disulfide bridge results in increase in Km, decrease in kcat value, loss of activity at 58°C | Aspergillus niger |
C31G | site-directed mutagenesis, the mutant shows altered pH and temperature optima compared to the wild-type enzyme | Aspergillus niger |
C345G | removal of disulfide bridge results in decrease in kcat value, loss of activity at 58°C | Aspergillus niger |
C435G | site-directed mutagenesis, the mutant shows altered pH and temperature optima compared to the wild-type enzyme | Aspergillus niger |
C71S | site-directed mutagenesis, inactive mutant | Aspergillus niger |
C71S | removal of disulfide bridge results in loss of activity | Aspergillus niger |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.045 | - |
myo-inositol hexakisphosphate | mutant C435G, pH 2.5, 42°C | Aspergillus niger | |
0.045 | - |
myo-inositol-1,2,3,4,5,6-hexakisphosphate | pH 2.5, 42°C, recombinant mutant C435G | Aspergillus niger | |
0.056 | - |
myo-inositol hexakisphosphate | mutant C264G, pH 6.0, 37°C | Aspergillus niger | |
0.056 | - |
myo-inositol-1,2,3,4,5,6-hexakisphosphate | pH 6.0, 37°C, recombinant mutant C264G | Aspergillus niger | |
0.07 | - |
myo-inositol hexakisphosphate | wild-type, pH 5.0, 58°C | Aspergillus niger | |
0.07 | - |
myo-inositol-1,2,3,4,5,6-hexakisphosphate | pH 5.0, 58°C, recombinant wild-type enzyme | Aspergillus niger | |
0.145 | - |
myo-inositol hexakisphosphate | mutant C215S, pH 5.0, 37°C | Aspergillus niger | |
0.145 | - |
myo-inositol-1,2,3,4,5,6-hexakisphosphate | pH 5.0, 37°C, recombinant mutant C215S | Aspergillus niger | |
0.18 | - |
myo-inositol hexakisphosphate | mutant C31G, pH 5.5, 53°C | Aspergillus niger | |
0.18 | - |
myo-inositol-1,2,3,4,5,6-hexakisphosphate | pH 5.5, 53°C, recombinant mutant C31G | Aspergillus niger |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
myo-inositol-1,2,3,4,5,6-hexakisphosphate + H2O | Aspergillus niger | - |
? + phosphate | - |
? | |
myo-inositol-1,2,3,4,5,6-hexakisphosphate + H2O | Aspergillus niger NRRL 3135 | - |
? + phosphate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Aspergillus niger | - |
NRRL 3135 | - |
Aspergillus niger | P34752 | gene phyA | - |
Aspergillus niger NRRL 3135 | - |
NRRL 3135 | - |
Aspergillus niger NRRL 3135 | P34752 | gene phyA | - |
Purification (Comment) | Organism |
---|---|
recombinant wild-type enzyme mutant enzymes from Pichia pastoris strain X33 by ammonium sulfate fractionation, and two different steps of ion exchange chromatography | Aspergillus niger |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
myo-inositol hexakisphosphate + H2O | - |
Aspergillus niger | 1D-myo-inositol 1,2,4,5,6-pentakisphosphate + 1D-myo-inositol-1,2,5,6 tetrakisphosphate + phosphate | - |
? | |
myo-inositol hexakisphosphate + H2O | - |
Aspergillus niger NRRL 3135 | 1D-myo-inositol 1,2,4,5,6-pentakisphosphate + 1D-myo-inositol-1,2,5,6 tetrakisphosphate + phosphate | - |
? | |
myo-inositol-1,2,3,4,5,6-hexakisphosphate + H2O | - |
Aspergillus niger | ? + phosphate | - |
? | |
myo-inositol-1,2,3,4,5,6-hexakisphosphate + H2O | - |
Aspergillus niger NRRL 3135 | ? + phosphate | - |
? |
Synonyms | Comment | Organism |
---|---|---|
PhyA | - |
Aspergillus niger |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
mutants C215S, C264G | Aspergillus niger |
37 | - |
recombinant mutants C215S and C264G | Aspergillus niger |
42 | - |
mutant C435G | Aspergillus niger |
42 | - |
recombinant mutant C435G | Aspergillus niger |
53 | - |
mutant C31G | Aspergillus niger |
53 | - |
recombinant mutant C31G | Aspergillus niger |
58 | - |
wild-type | Aspergillus niger |
58 | - |
recombinant wild-type enzyme | Aspergillus niger |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
3 | 6 | myo-inositol hexakisphosphate | mutant C264G, pH 6.0, 37°C | Aspergillus niger | |
3 | 6 | myo-inositol-1,2,3,4,5,6-hexakisphosphate | pH 6.0, 37°C, recombinant mutant C264G | Aspergillus niger | |
62 | - |
myo-inositol hexakisphosphate | mutant C215S, pH 5.0, 37°C | Aspergillus niger | |
62 | - |
myo-inositol-1,2,3,4,5,6-hexakisphosphate | pH 5.0, 37°C, recombinant mutant C215S | Aspergillus niger | |
90 | - |
myo-inositol hexakisphosphate | mutant C435G, pH 2.5, 42°C | Aspergillus niger | |
90 | - |
myo-inositol-1,2,3,4,5,6-hexakisphosphate | pH 2.5, 42°C, recombinant mutant C435G | Aspergillus niger | |
234 | - |
myo-inositol hexakisphosphate | mutant C31G, pH 5.5, 53°C | Aspergillus niger | |
234 | - |
myo-inositol-1,2,3,4,5,6-hexakisphosphate | pH 5.5, 53°C, recombinant mutant C31G | Aspergillus niger | |
350 | - |
myo-inositol hexakisphosphate | wild-type, pH 5.0, 58°C | Aspergillus niger | |
350 | - |
myo-inositol-1,2,3,4,5,6-hexakisphosphate | pH 5.0, 58°C, recombinant wild-type enzyme | Aspergillus niger |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
2.5 | - |
mutant C435G | Aspergillus niger |
2.5 | - |
recombinant mutant C435G | Aspergillus niger |
5 | - |
wild-type, mutant C215S | Aspergillus niger |
5 | - |
recombinant wild-type enzyme and mutant C215S | Aspergillus niger |
5.5 | - |
mutant C31G | Aspergillus niger |
5.5 | - |
recombinant mutant C31G | Aspergillus niger |
6 | - |
mutant C264G | Aspergillus niger |
6 | - |
recombinant mutant C264G | Aspergillus niger |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
additional information | - |
pH profiles of wild-type and mutant phytases PhyA, overview | Aspergillus niger |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00043 | - |
myo-inositol-1,2,3,4,5,6-hexakisphosphate | pH 5.0, 37°C, recombinant mutant C215S | Aspergillus niger | |
0.00072 | - |
myo-inositol-1,2,3,4,5,6-hexakisphosphate | pH 6.0, 37°C, recombinant mutant C264G | Aspergillus niger | |
0.0013 | - |
myo-inositol-1,2,3,4,5,6-hexakisphosphate | pH 5.5, 53°C, recombinant mutant C31G | Aspergillus niger | |
0.002 | - |
myo-inositol-1,2,3,4,5,6-hexakisphosphate | pH 2.5, 42°C, recombinant mutant C435G | Aspergillus niger | |
0.005 | - |
myo-inositol-1,2,3,4,5,6-hexakisphosphate | pH 5.0, 58°C, recombinant wild-type enzyme | Aspergillus niger | |
430 | - |
myo-inositol hexakisphosphate | mutant C215S, pH 5.0, 37°C | Aspergillus niger | |
720 | - |
myo-inositol hexakisphosphate | mutant C264G, pH 6.0, 37°C | Aspergillus niger | |
1300 | - |
myo-inositol hexakisphosphate | mutant C31G, pH 5.5, 53°C | Aspergillus niger | |
2000 | - |
myo-inositol hexakisphosphate | mutant C435G, pH 2.5, 42°C | Aspergillus niger | |
5000 | - |
myo-inositol hexakisphosphate | wild-type, pH 5.0, 58°C | Aspergillus niger |