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Literature summary for 3.1.3.8 extracted from
- Comparative study on different expression hosts for alkaline phytase engineered in Escherichia coli (2016), Appl. Biochem. Biotechnol., 179, 997-1010 .
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| gene phy168, recombinant expression of wild-type and mutant enzymes in Bacillus subtilis strain WB800N, Escherichia coli TOP10 cells or strain BL21, and Pichia pastoris strain GS115, subcloning in Escherichia coli, evaluation of the best expression systems, overview. High glycosylation degree of the proteins when expressed in Pichia pastoris strain GS115, when the expression host is changed from Escherichia coli to Pichia pastoris, deglycosylation is conducted. The enzyme expressed in Pichia pastoris is secreted. Wild-type enzyme and the mutants D24G, D24G/K70R/K111E/N121S secreted by Pichia pastoris show decreased thermostability compared with those expressed in Escherichia coli, correspondingly. This reduction in residual activity might be ascribed to the N-glycosylation of phy168 in Pichia pastoris | Bacillus subtilis |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| D24G | site-directed mutagenesis, the Bacillus subtilis-expressed variant D24G shows 62.7, 68.3, and 17.9% higher specific activity than those expressed in Escherichia coli at pH 7.0, 60°C, pH 7.0, 37°C, and pH 4.5, 37°C, respectively. The specific activity of the Escherichia coli-expressed D24G at pH 7.0 and 37°C is improved by 40.3 % as compared with that of the wild-type, while the improvement is 91.9% in Bacillus subtilis and 82.2% in Pichia pastoris, respectively. Overall, the mutant's activity is increased compared to the wild-type | Bacillus subtilis |
| D24G/K70R/K111E/N121S | site-directed mutagenesis, the Bacillus subtilis-expressed variant D24G/K70R/K111E/N121S shows 54.1, 42.6, and 10.8% higher specific activity than those expressed in Escherichia coli at pH 7.0, 60°C, pH 7.0, 37°C, and pH 4.5, 37°C, respectively. Overall, the mutant's activity is significantly increased compared to the wild-type | Bacillus subtilis |
| K265E | site-directed mutagenesis, the mutant shows similar activity compared to wild-type | Bacillus subtilis |
| additional information | the specific activities of the Bacillus subtilis-expressed phy168 proteins are mostly higher than those of the corresponding phy168 enzymes expressed in Escherichia coli. The activties of wild-type and mutant enzymes vary dependent on conditions and expression system, overview | Bacillus subtilis |
| S51A | site-directed mutagenesis. Overall, the mutant's activity is increased compared to the wild-type | Bacillus subtilis |
| S51A/K265E | site-directed mutagenesis, overall, the mutant's activity is increased compared to the wild-type | 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 | - |
- |
Posttranslational Modification
| Posttranslational Modification | Comment | Organism |
|---|---|---|
| glycoprotein | two potential N-glycosylated sites within the Asn-Xaa-Ser/Thr sequence are found in the phy168 phytase. High glycosylation degree of the proteins when expressed in Pichia pastoris strain GS115 | Bacillus subtilis |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 19.7 | - |
purified recombinant enzyme mutant D24G/K70R/K111E/N121S, expressed in Escherichia coli, pH 7.0, 60°C | Bacillus subtilis |
| 22.7 | - |
purified recombinant enzyme mutant D24G/K70R/K111E/N121S, expressed in Pichia pastoris, pH 7.0, 60°C | Bacillus subtilis |
| 30.4 | - |
purified recombinant enzyme mutant D24G/K70R/K111E/N121S, expressed in Bacillus subtilis, 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 | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| ? | x * 45000, Pichia pastoris-expressed Phy168 after deglycosylation, SDS-PAGE | Bacillus subtilis |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| phy168 | - |
Bacillus subtilis |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 60 | - |
recombinant wild-type enzyme | Bacillus subtilis |
Temperature Range [°C]
| Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
|---|---|---|---|
| additional information | - |
the highest activities of wild-type and D24G expressed in Pichia pastoris are achieved at 50°C and 55°C, decreased by 10°C and 5°C compared to those expressed in Escherichia coli, respectively. Temperature profiles of the mutants expressed in Pichia pastoris and Bacillus subtilis, overview | Bacillus subtilis |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| additional information | - |
temperature profiles of the mutants expressed in Pichia pastoris and Bacillus subtilis | Bacillus subtilis |
| 80 | - |
pH 7.0, 37°C, 70% activity remains for the purified recombinant enzyme mutant expressed from Bacillus subtilis, 25% activity remains for the purified recombinant enzyme mutant expressed from Pichia pastoris, and 50% activity remains for the purified recombinant enzyme mutant expressed from Escherichia coli | Bacillus subtilis |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7 | - |
recombinant wild-type enzyme | Bacillus subtilis |
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