Cloned (Comment) | Organism |
---|---|
gene appA or phyS, cloning from library, DNA and amino acid sequence determination and analysis, sequence comparisons and phylogenetic analysis, recombinant expression of extracellular His-tagged enzyme in Escherichia coli strain BL21(DE3) as nonglycosylated protein and of extracellular nontagged enzyme in Pichia pastoris strain GS115(his4) under control of the AOX1 promoter as glycosylated protein | Shigella sp. CD2 |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Cu2+ | - |
Shigella sp. CD2 | |
EDTA | - |
Shigella sp. CD2 | |
Fe2+ | - |
Shigella sp. CD2 | |
additional information | the recombinant enzyme shows resistance to trypsin | Shigella sp. CD2 | |
Zn2+ | - |
Shigella sp. CD2 |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.18 | - |
myo-inositol hexakisphosphate | pH 5.5, 37°C, recombinant nonglycosylated enzyme expressed from Escherichia coli | Shigella sp. CD2 | |
0.22 | - |
myo-inositol hexakisphosphate | pH 5.5, 37°C, recombinant glycosylated enzyme expressed from Pichia pastoris | Shigella sp. CD2 |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
extracellular | - |
Shigella sp. CD2 | - |
- |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Ca2+ | activates | Shigella sp. CD2 | |
Mg2+ | activates | Shigella sp. CD2 | |
Mn2+ | activates | Shigella sp. CD2 |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
myo-inositol hexakisphosphate + H2O | Shigella sp. CD2 | phosphate cleavage position is not determined, cf. EC 3.1.3.8, 3.1.3.26, and 3.1.3.72 | 1D-myo-inositol pentakisphosphate + phosphate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Shigella sp. CD2 | F6IQ86 | isolated from wheat rhizosphere | - |
Posttranslational Modification | Comment | Organism |
---|---|---|
glycoprotein | enzyme AppAS contains three potential sites of N-glycosylation, recombinant nontagged enzyme from Pichia pastoris strain GS115(his4) is glycosylated, deglycosylation of rAppAP is carried out using Endo H deglycosylase | Shigella sp. CD2 |
Purification (Comment) | Organism |
---|---|
recombinant nonglycosylated His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, recombinant glycosylated nontagged enzyme from Pichia pastoris strain GS115(his4) by cation exchange chromatography | Shigella sp. CD2 |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
967 | - |
purified recombinant glycosylated enzyme expressed from Pichia pastoris, pH 5.5, 37°C | Shigella sp. CD2 |
2982 | - |
purified recombinant nonglycosylated enzyme expressed from Escherichia coli, pH 5.5, 37°C | Shigella sp. CD2 |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | the enzyme shows no or poor activity with ATP, ADP, 4-nitrophenyl phosphate, diphosphate, glucose 6-phosphate, and fructose 6-phosphate | Shigella sp. CD2 | ? | - |
? | |
myo-inositol hexakisphosphate + H2O | phosphate cleavage position is not determined, cf. EC 3.1.3.8, 3.1.3.26, and 3.1.3.72 | Shigella sp. CD2 | 1D-myo-inositol pentakisphosphate + phosphate | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 59000-65000, recombinant glycosylated enzyme from Pichia pastoris, SDS-PAGE, x * 45000, recombinant deglycosylated enzyme from Pichia pastoris, SDS-PAGE | Shigella sp. CD2 |
More | AppAS contains three potential sites of N-glycosylation, a putative signal peptide of 22 amino acids at N-terminal end and 8 cysteine residues among which 99-130, 200-210, 404-413 are the most possible disulfide bond pairs. The calculated molecular mass of the protein with and without the signal sequence are about 47 and 45 kDa, respectively | Shigella sp. CD2 |
Synonyms | Comment | Organism |
---|---|---|
AppA | - |
Shigella sp. CD2 |
AppAS | - |
Shigella sp. CD2 |
More | cf. EC 3.1.3.8 and 3.1.3.72 | Shigella sp. CD2 |
phyS | - |
Shigella sp. CD2 |
phytase | - |
Shigella sp. CD2 |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
60 | - |
recombinant enzyme | Shigella sp. CD2 |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
10 | 80 | and below, activity profile overview, recombinant enzymes | Shigella sp. CD2 |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
720 | - |
myo-inositol hexakisphosphate | pH 5.5, 37°C, recombinant glycosylated enzyme expressed from Pichia pastoris | Shigella sp. CD2 | |
2230 | - |
myo-inositol hexakisphosphate | pH 5.5, 37°C, recombinant nonglycosylated enzyme expressed from Escherichia coli | Shigella sp. CD2 |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
5.5 | - |
recombinant enzyme | Shigella sp. CD2 |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
2.5 | 8.5 | activity range, profile overview, over 50% of activity at pH 3.5-6.5 | Shigella sp. CD2 |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
3272.7 | - |
myo-inositol hexakisphosphate | pH 5.5, 37°C, recombinant glycosylated enzyme expressed from Pichia pastoris | Shigella sp. CD2 | |
12389 | - |
myo-inositol hexakisphosphate | pH 5.5, 37°C, recombinant nonglycosylated enzyme expressed from Escherichia coli | Shigella sp. CD2 |