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Literature summary extracted from
- Enhanced activity of an alkaline phytase from Bacillus subtilis 168 in acidic and neutral environments by directed evolution (2015), Biochem. Eng. J., 98, 137-143 .
No PubMed abstract available
Cloned(Commentary)
| EC Number | Cloned (Comment) | Organism |
|---|---|---|
| 3.1.3.8 | gene phy, recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21 | Bacillus subtilis |
Protein Variants
| EC Number | Protein Variants | Comment | Organism |
|---|---|---|---|
| 3.1.3.8 | D24G | site-directed mutagenesis, the mutant shows 29.7% activity compared to the wild-type at pH 7.0, 60°C, and 76.6% at pH 4.5, 37°C | Bacillus subtilis |
| 3.1.3.8 | D24G/K111E | site-directed mutagenesis, the mutant shows increased activity compared to wild-type at pH 7.0, 60°C | Bacillus subtilis |
| 3.1.3.8 | D24G/K265E | site-directed mutagenesis, the mutant shows increased activity compared to wild-type at pH 7.0, 60°C | Bacillus subtilis |
| 3.1.3.8 | D24G/K265N | site-directed mutagenesis, the mutant shows 42.7% activity compared to the wild-type at pH 7.0, 60°C, and 84.2% at pH 4.5, 37°C | Bacillus subtilis |
| 3.1.3.8 | D24G/K265N | site-directed mutagenesis, the mutant shows increased activity compared to wild-type at pH 7.0, 60°C | Bacillus subtilis |
| 3.1.3.8 | D24G/K70R | site-directed mutagenesis, the mutant shows increased activity compared to wild-type at pH 7.0, 60°C | Bacillus subtilis |
| 3.1.3.8 | D24G/K70R/K111E/N121S | site-directed mutagenesis, the mutant shows 42.8% activity compared to the wild-type at pH 7.0, 60°C, and 121.1% at pH 4.5, 37°C | Bacillus subtilis |
| 3.1.3.8 | D24G/N121S | site-directed mutagenesis, the mutant shows increased activity compared to wild-type at pH 7.0, 60°C | Bacillus subtilis |
| 3.1.3.8 | D24G/S51A | site-directed mutagenesis, the mutant shows slightly decreased activity compared to wild-type at pH 7.0, 60°C | Bacillus subtilis |
| 3.1.3.8 | D24G/S51A/K265E | site-directed mutagenesis, the mutant shows increased activity compared to wild-type at pH 7.0, 60°C | Bacillus subtilis |
| 3.1.3.8 | D24G/S51P | site-directed mutagenesis, inactive mutant | Bacillus subtilis |
| 3.1.3.8 | K265E | site-directed mutagenesis, the mutant shows increased activity compared to wild-type at pH 7.0, 60°C | Bacillus subtilis |
| 3.1.3.8 | additional information | directed evolution and library screening, pH and tempearture profiles of the mutant enzymes compared to the wild-type enzyme, detailed overview | Bacillus subtilis |
| 3.1.3.8 | S51A | site-directed mutagenesis, the mutant shows 13.5% activity compared to the wild-type at pH 7.0, 60°C, and 79.5% at pH 4.5, 37°C | Bacillus subtilis |
| 3.1.3.8 | S51A/K265E | site-directed mutagenesis, the mutant shows increased activity compared to wild-type at pH 7.0, 60°C | Bacillus subtilis |
KM Value [mM]
| EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 3.1.3.8 | additional information | - |
additional information | Lineweaver-Burk kinetics | Bacillus subtilis | |
| 3.1.3.8 | 0.39 | - |
myo-inositol hexakisphosphate | pH 4.5, 37°C, recombinant mutant D24G/K70R/K111E/N121S | Bacillus subtilis |  |
| 3.1.3.8 | 0.49 | - |
myo-inositol hexakisphosphate | pH 4.5, 37°C, recombinant mutant D24G/K265N | Bacillus subtilis | |
| 3.1.3.8 | 0.54 | - |
myo-inositol hexakisphosphate | pH 4.5, 37°C, recombinant mutant D24G | Bacillus subtilis | |
| 3.1.3.8 | 0.78 | - |
myo-inositol hexakisphosphate | pH 4.5, 37°C, recombinant wild-type enzyme | Bacillus subtilis | |
| 3.1.3.8 | 1.4 | - |
myo-inositol hexakisphosphate | pH 7.0, 37°C, recombinant mutant D24G | Bacillus subtilis | |
| 3.1.3.8 | 1.5 | - |
myo-inositol hexakisphosphate | pH 7.0, 37°C, recombinant mutant D24G/K70R/K111E/N121S | Bacillus subtilis | |
| 3.1.3.8 | 1.77 | - |
myo-inositol hexakisphosphate | pH 7.0, 37°C, recombinant mutant D24G/K265N | Bacillus subtilis | |
| 3.1.3.8 | 2.19 | - |
myo-inositol hexakisphosphate | pH 7.0, 37°C, recombinant wild-type enzyme | Bacillus subtilis | |
Natural Substrates/ Products (Substrates)
| EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 3.1.3.8 | myo-inositol hexakisphosphate + H2O | Bacillus subtilis | - |
1D-myo-inositol 1,2,4,5,6-pentakisphosphate + phosphate | - |
? | |
| 3.1.3.8 | myo-inositol hexakisphosphate + H2O | Bacillus subtilis 168 | - |
1D-myo-inositol 1,2,4,5,6-pentakisphosphate + phosphate | - |
? | |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 3.1.3.8 | Bacillus subtilis | P42094 | - |
- |
| 3.1.3.8 | Bacillus subtilis 168 | P42094 | - |
- |
Purification (Commentary)
| EC Number | Purification (Comment) | Organism |
|---|---|---|
| 3.1.3.8 | recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21 by nickel affinity chromatography | Bacillus subtilis |
Specific Activity [micromol/min/mg]
| EC Number | Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|---|
| 3.1.3.8 | 1.71 | - |
purified recombinant His-tagged wild-type enzyme, pH 4.5, 37°C | Bacillus subtilis |
| 3.1.3.8 | 6.32 | - |
purified recombinant His-tagged wild-type enzyme, pH 7.0, 37°C | Bacillus subtilis |
| 3.1.3.8 | 12.81 | - |
purified recombinant His-tagged mutant K265E, pH 7.0, 60°C | Bacillus subtilis |
| 3.1.3.8 | 13.82 | - |
purified recombinant His-tagged wild-type enzyme, pH 7.0, 60°C | Bacillus subtilis |
| 3.1.3.8 | 15.68 | - |
purified recombinant His-tagged mutant S51A, pH 7.0, 60°C | Bacillus subtilis |
| 3.1.3.8 | 17.24 | - |
purified recombinant His-tagged mutant S51A/K265E, pH 7.0, 60°C | Bacillus subtilis |
| 3.1.3.8 | 17.93 | - |
purified recombinant His-tagged mutant D24G, pH 7.0, 60°C | Bacillus subtilis |
| 3.1.3.8 | 18.14 | - |
purified recombinant His-tagged mutant D24G/S51A, pH 7.0, 60°C | Bacillus subtilis |
| 3.1.3.8 | 18.17 | - |
purified recombinant His-tagged mutant D24G/N121S, pH 7.0, 60°C | Bacillus subtilis |
| 3.1.3.8 | 18.88 | - |
purified recombinant His-tagged mutant D24G/K111E, pH 7.0, 60°C | Bacillus subtilis |
| 3.1.3.8 | 19.27 | - |
purified recombinant His-tagged mutant D24G/K265E, pH 7.0, 60°C | Bacillus subtilis |
| 3.1.3.8 | 19.33 | - |
purified recombinant His-tagged mutant D24G/S51A/K265E, pH 7.0, 60°C | Bacillus subtilis |
| 3.1.3.8 | 19.67 | - |
purified recombinant His-tagged mutant D24G/K70R, pH 7.0, 60°C | Bacillus subtilis |
| 3.1.3.8 | 19.72 | - |
purified recombinant His-tagged mutant D24G/K265N, pH 7.0, 60°C | Bacillus subtilis |
| 3.1.3.8 | 19.73 | - |
purified recombinant His-tagged mutant D24G/K70R/K111E/N121S, pH 7.0, 60°C | Bacillus subtilis |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 3.1.3.8 | myo-inositol hexakisphosphate + H2O | - |
Bacillus subtilis | 1D-myo-inositol 1,2,4,5,6-pentakisphosphate + phosphate | - |
? | |
| 3.1.3.8 | myo-inositol hexakisphosphate + H2O | - |
Bacillus subtilis 168 | 1D-myo-inositol 1,2,4,5,6-pentakisphosphate + phosphate | - |
? | |
Synonyms
| EC Number | Synonyms | Comment | Organism |
|---|---|---|---|
| 3.1.3.8 | alkaline phytase | - |
Bacillus subtilis |
| 3.1.3.8 | Phy | - |
Bacillus subtilis |
| 3.1.3.8 | phy168 | - |
Bacillus subtilis |
Temperature Optimum [°C]
| EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 3.1.3.8 | 60 | - |
- |
Bacillus subtilis |
Turnover Number [1/s]
| EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 3.1.3.8 | 1837.8 | - |
myo-inositol hexakisphosphate | pH 4.5, 37°C, recombinant wild-type enzyme | Bacillus subtilis | |
| 3.1.3.8 | 2450.5 | - |
myo-inositol hexakisphosphate | pH 4.5, 37°C, recombinant mutant D24G/K70R/K111E/N121S | Bacillus subtilis | |
| 3.1.3.8 | 2510.3 | - |
myo-inositol hexakisphosphate | pH 4.5, 37°C, recombinant mutant D24G/K265N | Bacillus subtilis | |
| 3.1.3.8 | 2527.8 | - |
myo-inositol hexakisphosphate | pH 4.5, 37°C, recombinant mutant D24G | Bacillus subtilis | |
| 3.1.3.8 | 4965.7 | - |
myo-inositol hexakisphosphate | pH 7.0, 37°C, recombinant wild-type enzyme | Bacillus subtilis | |
| 3.1.3.8 | 7367.6 | - |
myo-inositol hexakisphosphate | pH 7.0, 37°C, recombinant mutant D24G | Bacillus subtilis | |
| 3.1.3.8 | 7860 | - |
myo-inositol hexakisphosphate | pH 7.0, 37°C, recombinant mutant D24G/K70R/K111E/N121S | Bacillus subtilis | |
| 3.1.3.8 | 8430.1 | - |
myo-inositol hexakisphosphate | pH 7.0, 37°C, recombinant mutant D24G/K265N | Bacillus subtilis | |
pH Optimum
| EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|---|
| 3.1.3.8 | 7 | - |
- |
Bacillus subtilis |
General Information
| EC Number | General Information | Comment | Organism |
|---|---|---|---|
| 3.1.3.8 | additional information | molecular docking of wild-type and mutant enzymes, homology modelling using the enzyme structure of the phytase from Bacillus amyloliquefaciens (PDB ID 2POO) as the template, overview. Residues D24, S51 and K70 have a relatively large influence on the catalytic activity probably because they are adjacent to the active site cleft. The enzyme is composed of six beta-sheets and possesses six calcium binding sites. The 25 amino acid residues except V60, involved in calcium binding, are highly conserved | Bacillus subtilis |
kcat/KM [mM/s]
| EC Number | kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 3.1.3.8 | 2267.5 | - |
myo-inositol hexakisphosphate | pH 7.0, 37°C, recombinant wild-type enzyme | Bacillus subtilis | |
| 3.1.3.8 | 2368.3 | - |
myo-inositol hexakisphosphate | pH 4.5, 37°C, recombinant wild-type enzyme | Bacillus subtilis | |
| 3.1.3.8 | 4695.6 | - |
myo-inositol hexakisphosphate | pH 4.5, 37°C, recombinant mutant D24G | Bacillus subtilis | |
| 3.1.3.8 | 4767.2 | - |
myo-inositol hexakisphosphate | pH 7.0, 37°C, recombinant mutant D24G/K265N | Bacillus subtilis | |
| 3.1.3.8 | 5080.3 | - |
myo-inositol hexakisphosphate | pH 4.5, 37°C, recombinant mutant D24G/K265N | Bacillus subtilis | |
| 3.1.3.8 | 5231.8 | - |
myo-inositol hexakisphosphate | pH 7.0, 37°C, recombinant mutant D24G/K70R/K111E/N121S | Bacillus subtilis | |
| 3.1.3.8 | 5258.8 | - |
myo-inositol hexakisphosphate | pH 7.0, 37°C, recombinant mutant D24G | Bacillus subtilis | |
| 3.1.3.8 | 6235.5 | - |
myo-inositol hexakisphosphate | pH 4.5, 37°C, recombinant mutant D24G/K70R/K111E/N121S | Bacillus subtilis | |
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