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Literature summary extracted from

  • Pal Roy, M.; Mazumdar, D.; Dutta, S.; Saha, S.P.; Ghosh, S.
    Cloning and expression of phytase appA gene from Shigella sp. CD2 in Pichia pastoris and comparison of properties with recombinant enzyme expressed in E. coli (2016), PLoS ONE, 11, e0145745 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.1.3.8 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
3.1.3.26 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
3.1.3.72 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 non-glycosylated 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

EC Number Inhibitors Comment Organism Structure
3.1.3.8 Cu2+
-
 Shigella sp. CD2
3.1.3.8 EDTA
-
 Shigella sp. CD2
3.1.3.8 Fe2+
-
 Shigella sp. CD2
3.1.3.8 additional information the recombinant enzyme shows resistance to trypsin Shigella sp. CD2
3.1.3.8 Zn2+
-
 Shigella sp. CD2
3.1.3.26 Cu2+
-
 Shigella sp. CD2
3.1.3.26 EDTA
-
 Shigella sp. CD2
3.1.3.26 Fe2+
-
 Shigella sp. CD2
3.1.3.26 additional information the recombinant enzyme shows resistance to trypsin Shigella sp. CD2
3.1.3.26 Zn2+
-
 Shigella sp. CD2
3.1.3.72 Cu2+
-
 Shigella sp. CD2
3.1.3.72 EDTA
-
 Shigella sp. CD2
3.1.3.72 Fe2+
-
 Shigella sp. CD2
3.1.3.72 additional information the recombinant enzyme shows resistance to trypsin Shigella sp. CD2
3.1.3.72 Zn2+
-
 Shigella sp. CD2

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.1.3.8 0.18
-
 myo-inositol hexakisphosphate pH 5.5, 37°C, recombinant nonglycosylated enzyme expressed from Escherichia coli Shigella sp. CD2
3.1.3.8 0.22
-
 myo-inositol hexakisphosphate pH 5.5, 37°C, recombinant glycosylated enzyme expressed from Pichia pastoris Shigella sp. CD2
3.1.3.26 0.18
-
 myo-inositol hexakisphosphate pH 5.5, 37°C, recombinant nonglycosylated enzyme expressed from Escherichia coli Shigella sp. CD2
3.1.3.26 0.22
-
 myo-inositol hexakisphosphate pH 5.5, 37°C, recombinant glycosylated enzyme expressed from Pichia pastoris Shigella sp. CD2
3.1.3.72 0.18
-
 myo-inositol hexakisphosphate pH 5.5, 37°C, recombinant non-glycosylated enzyme expressed from Escherichia coli Shigella sp. CD2
3.1.3.72 0.22
-
 myo-inositol hexakisphosphate pH 5.5, 37°C, recombinant glycosylated enzyme expressed from Pichia pastoris Shigella sp. CD2

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
3.1.3.8 extracellular
-
 Shigella sp. CD2
-
-
3.1.3.26 extracellular
-
 Shigella sp. CD2
-
-
3.1.3.72 extracellular
-
 Shigella sp. CD2
-
-

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.1.3.8 Ca2+ activates Shigella sp. CD2
3.1.3.8 Mg2+ activates Shigella sp. CD2
3.1.3.8 Mn2+ activates Shigella sp. CD2
3.1.3.26 Ca2+ activates Shigella sp. CD2
3.1.3.26 Mg2+ activates Shigella sp. CD2
3.1.3.26 Mn2+ activates Shigella sp. CD2
3.1.3.72 Ca2+ activates Shigella sp. CD2
3.1.3.72 Mg2+ activates Shigella sp. CD2
3.1.3.72 Mn2+ activates Shigella sp. CD2

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 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
-
 ?
3.1.3.26 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
-
 ?
3.1.3.72 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

EC Number Organism UniProt Comment Textmining
3.1.3.8 Shigella sp. CD2 F6IQ86 isolated from wheat rhizosphere
-
3.1.3.26 Shigella sp. CD2 F6IQ86 isolated from wheat rhizosphere
-
3.1.3.72 Shigella sp. CD2 F6IQ86 isolated from wheat rhizosphere
-

Posttranslational Modification

EC Number Posttranslational Modification Comment Organism
3.1.3.8 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
3.1.3.26 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
3.1.3.72 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 (Commentary)

EC Number Purification (Comment) Organism
3.1.3.8 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
3.1.3.26 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
3.1.3.72 recombinant non-glycosylated 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 [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.1.3.8 967
-
 purified recombinant glycosylated enzyme expressed from Pichia pastoris, pH 5.5, 37°C Shigella sp. CD2
3.1.3.8 2982
-
 purified recombinant nonglycosylated enzyme expressed from Escherichia coli, pH 5.5, 37°C Shigella sp. CD2
3.1.3.26 967
-
 purified recombinant glycosylated enzyme expressed from Pichia pastoris, pH 5.5, 37°C Shigella sp. CD2
3.1.3.26 2982
-
 purified recombinant nonglycosylated enzyme expressed from Escherichia coli, pH 5.5, 37°C Shigella sp. CD2
3.1.3.72 967
-
 purified recombinant glycosylated enzyme expressed from Pichia pastoris, pH 5.5, 37°C Shigella sp. CD2
3.1.3.72 2982
-
 purified recombinant non-glycosylated enzyme expressed from Escherichia coli, pH 5.5, 37°C Shigella sp. CD2

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.1.3.8 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 ?
-
 ?
3.1.3.8 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
-
 ?
3.1.3.26 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 ?
-
 ?
3.1.3.26 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
-
 ?
3.1.3.72 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 ?
-
 ?
3.1.3.72 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

EC Number Subunits Comment Organism
3.1.3.8 ? x * 59000-65000, recombinant glycosylated enzyme from Pichia pastoris, SDS-PAGE, x * 45000, recombinant deglycosylated enzyme from Pichia pastoris, SDS-PAGE Shigella sp. CD2
3.1.3.8 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
3.1.3.26 ? x * 59000-65000, recombinant glycosylated enzyme from Pichia pastoris, SDS-PAGE, x * 45000, recombinant deglycosylated enzyme from Pichia pastoris, SDS-PAGE Shigella sp. CD2
3.1.3.26 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
3.1.3.72 ? x * 59000-65000, recombinant glycosylated enzyme from Pichia pastoris, SDS-PAGE, x * 45000, recombinant deglycosylated enzyme from Pichia pastoris, SDS-PAGE Shigella sp. CD2
3.1.3.72 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

EC Number Synonyms Comment Organism
3.1.3.8 AppA
-
 Shigella sp. CD2
3.1.3.8 AppAS
-
 Shigella sp. CD2
3.1.3.8 More cf. EC 3.1.3.26 and 3.1.3.72 Shigella sp. CD2
3.1.3.8 phyS
-
 Shigella sp. CD2
3.1.3.8 phytase
-
 Shigella sp. CD2
3.1.3.26 AppA
-
 Shigella sp. CD2
3.1.3.26 AppAS
-
 Shigella sp. CD2
3.1.3.26 More cf. EC 3.1.3.8 and 3.1.3.72 Shigella sp. CD2
3.1.3.26 phyS
-
 Shigella sp. CD2
3.1.3.26 phytase
-
 Shigella sp. CD2
3.1.3.72 AppA
-
 Shigella sp. CD2
3.1.3.72 AppAS
-
 Shigella sp. CD2
3.1.3.72 More cf. EC 3.1.3.8 and 3.1.3.26 Shigella sp. CD2
3.1.3.72 phyS
-
 Shigella sp. CD2
3.1.3.72 phytase
-
 Shigella sp. CD2

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.1.3.8 60
-
 recombinant enzyme Shigella sp. CD2
3.1.3.26 60
-
 recombinant enzyme Shigella sp. CD2
3.1.3.72 60
-
 recombinant enzyme Shigella sp. CD2

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
3.1.3.8 10 80 and below, activity profile overview, recombinant enzymes Shigella sp. CD2
3.1.3.26 10 80 and below, activity profile overview, recombinant enzymes Shigella sp. CD2
3.1.3.72 10 80 and below, activity profile overview, recombinant enzymes Shigella sp. CD2

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.1.3.8 720
-
 myo-inositol hexakisphosphate pH 5.5, 37°C, recombinant glycosylated enzyme expressed from Pichia pastoris Shigella sp. CD2
3.1.3.8 2230
-
 myo-inositol hexakisphosphate pH 5.5, 37°C, recombinant nonglycosylated enzyme expressed from Escherichia coli Shigella sp. CD2
3.1.3.26 720
-
 myo-inositol hexakisphosphate pH 5.5, 37°C, recombinant glycosylated enzyme expressed from Pichia pastoris Shigella sp. CD2
3.1.3.26 2230
-
 myo-inositol hexakisphosphate pH 5.5, 37°C, recombinant nonglycosylated enzyme expressed from Escherichia coli Shigella sp. CD2
3.1.3.72 720
-
 myo-inositol hexakisphosphate pH 5.5, 37°C, recombinant glycosylated enzyme expressed from Pichia pastoris Shigella sp. CD2
3.1.3.72 2230
-
 myo-inositol hexakisphosphate pH 5.5, 37°C, recombinant non-glycosylated enzyme expressed from Escherichia coli Shigella sp. CD2

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.1.3.8 5.5
-
 recombinant enzyme Shigella sp. CD2
3.1.3.26 5.5
-
 recombinant enzyme Shigella sp. CD2
3.1.3.72 5.5
-
 recombinant enzyme Shigella sp. CD2

pH Range

EC Number pH Minimum pH Maximum Comment Organism
3.1.3.8 2.5 8.5 activity range, profile overview, over 50% of activity at pH 3.5-6.5 Shigella sp. CD2
3.1.3.26 2.5 8.5 activity range, profile overview, over 50% of activity at pH 3.5-6.5 Shigella sp. CD2
3.1.3.72 2.5 8.5 activity range, profile overview, over 50% of activity at pH 3.5-6.5 Shigella sp. CD2

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 3272.7
-
 myo-inositol hexakisphosphate pH 5.5, 37°C, recombinant glycosylated enzyme expressed from Pichia pastoris Shigella sp. CD2
3.1.3.8 12389
-
 myo-inositol hexakisphosphate pH 5.5, 37°C, recombinant nonglycosylated enzyme expressed from Escherichia coli Shigella sp. CD2
3.1.3.26 3272.7
-
 myo-inositol hexakisphosphate pH 5.5, 37°C, recombinant glycosylated enzyme expressed from Pichia pastoris Shigella sp. CD2
3.1.3.26 12389
-
 myo-inositol hexakisphosphate pH 5.5, 37°C, recombinant nonglycosylated enzyme expressed from Escherichia coli Shigella sp. CD2
3.1.3.72 3272.7
-
 myo-inositol hexakisphosphate pH 5.5, 37°C, recombinant glycosylated enzyme expressed from Pichia pastoris Shigella sp. CD2
3.1.3.72 12389
-
 myo-inositol hexakisphosphate pH 5.5, 37°C, recombinant non-glycosylated enzyme expressed from Escherichia coli Shigella sp. CD2