Cloned (Comment) | Organism |
---|---|
expressed in Pichia pastoris X-33 | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
D144N/V227A | 38% higher specific activity than wild-type, lower efficiency for soy phytate hydrolysis compared to wild-type enzyme, mutant shows higher thermostability compared to wild-type, Km (myo-inositol hexakisphosphate) lower compared to wild-type, kcat similar to wild-type | Escherichia coli |
D144N/V227A/G344D | mutant releases 25% more inorganic phosphorus from soy phytate than the wild-type enzyme, mutant shows higher thermostability compared to wild-type, Km (myo-inositol hexakisphosphate) lower compared to wild-type, kcat similar to wild-type | Escherichia coli |
D144N/V227A/G344D/D112N | 7% higher specific activity than wild-type, mutant shows higher thermostability compared to wild-type | Escherichia coli |
D144N/V227A/G344D/D112N/K46E | lower efficiency for soy phytate hydrolysis compared to wild-type enzyme, mutant shows higher thermostability compared to wild-type | Escherichia coli |
D144N/V227A/G344D/D112N/K46E/G103S | lower efficiency for soy phytate hydrolysis compared to wild-type enzyme, mutant shows higher thermostability compared to wild-type | Escherichia coli |
D144N/V227A/G344D/D112N/K46E/G103S/S209G | 23% higher specific activity than wild-type, lower efficiency for soy phytate hydrolysis compared to wild-type enzyme | Escherichia coli |
D144N/V227A/G344D/K46E | similar efficiency for soy phytate hydrolysis as compared to wild-type enzyme, mutant shows higher thermostability compared to wild-type | Escherichia coli |
D144N/V227A/G344D/K65E | lower efficiency for soy phytate hydrolysis compared to wild-type enzyme, mutant shows higher thermostability compared to wild-type | Escherichia coli |
additional information | no further improvement in thermostability is observed by adding other mutations to D144N/V227A/G344D, which might result from unfavorable electrostatic interactions or structural perturbation | Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0318 | - |
myo-inositol hexakisphosphate | mutant D144N/V227A/G344D | Escherichia coli | |
0.04875 | - |
myo-inositol hexakisphosphate | mutant D144N/V227A | Escherichia coli | |
0.1235 | - |
myo-inositol hexakisphosphate | wild-type | Escherichia coli |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | Q6RK08 | the enzyme may be a 3-phytase, EC 3.1.3.8, or a 4-phytase (synonym 6-phytase, EC 3.1.3.26). The product of the hydrolysis of myo-inositol hexakisphosphate to 1D-myo-inositol 1,2,4,5,6-pentakisphosphate (3-phytase) or 1D-myo-inositol 1,2,3,5,6-pentakisphosphate (4-phytase) (i.e. 1L-myo-inositol 1,2,3,4,5-pentakisphosphate if 1L numbering is applied) has not been analyzed. The reaction was monitored by analyzing the released phosphate | - |
Purification (Comment) | Organism |
---|---|
purified by ultrafiltration and Macro-Prep high S-cation exchange chromatography | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
myo-inositol hexakisphosphate + H2O | The enzyme may be a 3-phytase, EC 3.1.3.8, or a 4-phytase (synonym 6-phytase, EC 3.1.3.26). The product of the hydrolysis of myo-inositol hexakisphosphate to 1D-myo-inositol 1,2,4,5,6-pentakisphosphate (3-phytase) or 1D-myo-inositol 1,2,3,5,6-pentakisphosphate (4-phytase) (i.e. 1L-myo-inositol 1,2,3,4,5-pentakisphosphate if 1L numbering is applied) has not been analyzed. The reaction was monitored by analyzing the released phosphate | Escherichia coli | ? + phosphate | - |
? |
Synonyms | Comment | Organism |
---|---|---|
AppA2 phytase | - |
Escherichia coli |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Escherichia coli |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.87 | - |
myo-inositol hexakisphosphate | mutant D144N/V227A/G344D | Escherichia coli | |
0.94 | - |
myo-inositol hexakisphosphate | wild-type | Escherichia coli | |
0.97 | - |
myo-inositol hexakisphosphate | mutant D144N/V227A | Escherichia coli |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
3.5 | - |
assay at | Escherichia coli |