Any feedback?
Literature summary for 3.1.3.8 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)
| Cloned (Comment) | Organism |
|---|---|
| gene phy, recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21 | Bacillus subtilis |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| 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 |
| D24G/K111E | site-directed mutagenesis, the mutant shows increased activity compared to wild-type at pH 7.0, 60°C | Bacillus subtilis |
| D24G/K265E | site-directed mutagenesis, the mutant shows increased activity compared to wild-type at pH 7.0, 60°C | Bacillus subtilis |
| 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 |
| D24G/K265N | site-directed mutagenesis, the mutant shows increased activity compared to wild-type at pH 7.0, 60°C | Bacillus subtilis |
| D24G/K70R | site-directed mutagenesis, the mutant shows increased activity compared to wild-type at pH 7.0, 60°C | Bacillus subtilis |
| 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 |
| D24G/N121S | site-directed mutagenesis, the mutant shows increased activity compared to wild-type at pH 7.0, 60°C | Bacillus subtilis |
| D24G/S51A | site-directed mutagenesis, the mutant shows slightly decreased activity compared to wild-type at pH 7.0, 60°C | Bacillus subtilis |
| D24G/S51A/K265E | site-directed mutagenesis, the mutant shows increased activity compared to wild-type at pH 7.0, 60°C | Bacillus subtilis |
| D24G/S51P | site-directed mutagenesis, inactive mutant | Bacillus subtilis |
| K265E | site-directed mutagenesis, the mutant shows increased activity compared to wild-type at pH 7.0, 60°C | Bacillus subtilis |
| 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 |
| 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 |
| 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]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| additional information | - |
additional information | Lineweaver-Burk kinetics | Bacillus subtilis | |
| 0.39 | - |
myo-inositol hexakisphosphate | pH 4.5, 37°C, recombinant mutant D24G/K70R/K111E/N121S | Bacillus subtilis |  |
| 0.49 | - |
myo-inositol hexakisphosphate | pH 4.5, 37°C, recombinant mutant D24G/K265N | Bacillus subtilis | |
| 0.54 | - |
myo-inositol hexakisphosphate | pH 4.5, 37°C, recombinant mutant D24G | Bacillus subtilis | |
| 0.78 | - |
myo-inositol hexakisphosphate | pH 4.5, 37°C, recombinant wild-type enzyme | Bacillus subtilis | |
| 1.4 | - |
myo-inositol hexakisphosphate | pH 7.0, 37°C, recombinant mutant D24G | Bacillus subtilis | |
| 1.5 | - |
myo-inositol hexakisphosphate | pH 7.0, 37°C, recombinant mutant D24G/K70R/K111E/N121S | Bacillus subtilis | |
| 1.77 | - |
myo-inositol hexakisphosphate | pH 7.0, 37°C, recombinant mutant D24G/K265N | Bacillus subtilis | |
| 2.19 | - |
myo-inositol hexakisphosphate | pH 7.0, 37°C, recombinant wild-type enzyme | Bacillus subtilis | |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| myo-inositol hexakisphosphate + H2O | Bacillus subtilis | - |
1D-myo-inositol 1,2,4,5,6-pentakisphosphate + phosphate | - |
? | |
| myo-inositol hexakisphosphate + H2O | Bacillus subtilis 168 | - |
1D-myo-inositol 1,2,4,5,6-pentakisphosphate + phosphate | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Bacillus subtilis | P42094 | - |
- |
| Bacillus subtilis 168 | P42094 | - |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21 by nickel affinity chromatography | Bacillus subtilis |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 1.71 | - |
purified recombinant His-tagged wild-type enzyme, pH 4.5, 37°C | Bacillus subtilis |
| 6.32 | - |
purified recombinant His-tagged wild-type enzyme, pH 7.0, 37°C | Bacillus subtilis |
| 12.81 | - |
purified recombinant His-tagged mutant K265E, pH 7.0, 60°C | Bacillus subtilis |
| 13.82 | - |
purified recombinant His-tagged wild-type enzyme, pH 7.0, 60°C | Bacillus subtilis |
| 15.68 | - |
purified recombinant His-tagged mutant S51A, pH 7.0, 60°C | Bacillus subtilis |
| 17.24 | - |
purified recombinant His-tagged mutant S51A/K265E, pH 7.0, 60°C | Bacillus subtilis |
| 17.93 | - |
purified recombinant His-tagged mutant D24G, pH 7.0, 60°C | Bacillus subtilis |
| 18.14 | - |
purified recombinant His-tagged mutant D24G/S51A, pH 7.0, 60°C | Bacillus subtilis |
| 18.17 | - |
purified recombinant His-tagged mutant D24G/N121S, pH 7.0, 60°C | Bacillus subtilis |
| 18.88 | - |
purified recombinant His-tagged mutant D24G/K111E, pH 7.0, 60°C | Bacillus subtilis |
| 19.27 | - |
purified recombinant His-tagged mutant D24G/K265E, pH 7.0, 60°C | Bacillus subtilis |
| 19.33 | - |
purified recombinant His-tagged mutant D24G/S51A/K265E, pH 7.0, 60°C | Bacillus subtilis |
| 19.67 | - |
purified recombinant His-tagged mutant D24G/K70R, pH 7.0, 60°C | Bacillus subtilis |
| 19.72 | - |
purified recombinant His-tagged mutant D24G/K265N, pH 7.0, 60°C | Bacillus subtilis |
| 19.73 | - |
purified recombinant His-tagged mutant D24G/K70R/K111E/N121S, pH 7.0, 60°C | Bacillus subtilis |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| myo-inositol hexakisphosphate + H2O | - |
Bacillus subtilis | 1D-myo-inositol 1,2,4,5,6-pentakisphosphate + phosphate | - |
? | |
| myo-inositol hexakisphosphate + H2O | - |
Bacillus subtilis 168 | 1D-myo-inositol 1,2,4,5,6-pentakisphosphate + phosphate | - |
? | |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| alkaline phytase | - |
Bacillus subtilis |
| Phy | - |
Bacillus subtilis |
| phy168 | - |
Bacillus subtilis |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 60 | - |
- |
Bacillus subtilis |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 1837.8 | - |
myo-inositol hexakisphosphate | pH 4.5, 37°C, recombinant wild-type enzyme | Bacillus subtilis | |
| 2450.5 | - |
myo-inositol hexakisphosphate | pH 4.5, 37°C, recombinant mutant D24G/K70R/K111E/N121S | Bacillus subtilis | |
| 2510.3 | - |
myo-inositol hexakisphosphate | pH 4.5, 37°C, recombinant mutant D24G/K265N | Bacillus subtilis | |
| 2527.8 | - |
myo-inositol hexakisphosphate | pH 4.5, 37°C, recombinant mutant D24G | Bacillus subtilis | |
| 4965.7 | - |
myo-inositol hexakisphosphate | pH 7.0, 37°C, recombinant wild-type enzyme | Bacillus subtilis | |
| 7367.6 | - |
myo-inositol hexakisphosphate | pH 7.0, 37°C, recombinant mutant D24G | Bacillus subtilis | |
| 7860 | - |
myo-inositol hexakisphosphate | pH 7.0, 37°C, recombinant mutant D24G/K70R/K111E/N121S | Bacillus subtilis | |
| 8430.1 | - |
myo-inositol hexakisphosphate | pH 7.0, 37°C, recombinant mutant D24G/K265N | Bacillus subtilis | |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7 | - |
- |
Bacillus subtilis |
General Information
| General Information | Comment | Organism |
|---|---|---|
| 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]
| kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 2267.5 | - |
myo-inositol hexakisphosphate | pH 7.0, 37°C, recombinant wild-type enzyme | Bacillus subtilis | |
| 2368.3 | - |
myo-inositol hexakisphosphate | pH 4.5, 37°C, recombinant wild-type enzyme | Bacillus subtilis | |
| 4695.6 | - |
myo-inositol hexakisphosphate | pH 4.5, 37°C, recombinant mutant D24G | Bacillus subtilis | |
| 4767.2 | - |
myo-inositol hexakisphosphate | pH 7.0, 37°C, recombinant mutant D24G/K265N | Bacillus subtilis | |
| 5080.3 | - |
myo-inositol hexakisphosphate | pH 4.5, 37°C, recombinant mutant D24G/K265N | Bacillus subtilis | |
| 5231.8 | - |
myo-inositol hexakisphosphate | pH 7.0, 37°C, recombinant mutant D24G/K70R/K111E/N121S | Bacillus subtilis | |
| 5258.8 | - |
myo-inositol hexakisphosphate | pH 7.0, 37°C, recombinant mutant D24G | Bacillus subtilis | |
| 6235.5 | - |
myo-inositol hexakisphosphate | pH 4.5, 37°C, recombinant mutant D24G/K70R/K111E/N121S | Bacillus subtilis | |
Use of this online version of BRENDA is free under the CC BY 4.0 license. See terms of use for full details.
Release 2023.1 (January 2023)
Legal
Curated at
Member of
