| IED ID | IndEnz0002011709 |
| Enzyme Type ID | protease011709 |
| Protein Name |
Phytase AppA EC 3.1.3.- 6-phytase Histidine acid phosphatase phytase HAP phytase Myo-inositol hexakisphosphate phosphohydrolase Phosphoanhydride phosphatase EC 3.6.1.- pH 2.5 acid phosphatase Acid phosphatase EC 3.1.3.- |
| Gene Name | appA b0980 JW0963 |
| Organism | Escherichia coli (strain K12) |
| Taxonomic Lineage | cellular organisms Bacteria Proteobacteria Gammaproteobacteria Enterobacterales Enterobacteriaceae Escherichia Escherichia coli Escherichia coli (strain K12) |
| Enzyme Sequence | MKAILIPFLSLLIPLTPQSAFAQSEPELKLESVVIVSRHGVRAPTKATQLMQDVTPDAWPTWPVKLGWLTPRGGELIAYLGHYQRQRLVADGLLAKKGCPQSGQVAIIADVDERTRKTGEAFAAGLAPDCAITVHTQADTSSPDPLFNPLKTGVCQLDNANVTDAILSRAGGSIADFTGHRQTAFRELERVLNFPQSNLCLKREKQDESCSLTQALPSELKVSADNVSLTGAVSLASMLTEIFLLQQAQGMPEPGWGRITDSHQWNTLLSLHNAQFYLLQRTPEVARSRATPLLDLIKTALTPHPPQKQAYGVTLPTSVLFIAGHDTNLANLGGALELNWTLPGQPDNTPPGGELVFERWRRLSDNSQWIQVSLVFQTLQQMRDKTPLSLNTPPGEVKLTLAGCEERNAQGMCSLAGFTQIVNEARIPACSL |
| Enzyme Length | 432 |
| Uniprot Accession Number | P07102 |
| Absorption | |
| Active Site | ACT_SITE 39; /note="Nucleophile"; /evidence="ECO:0000305|PubMed:10655611, ECO:0000305|PubMed:1429631"; ACT_SITE 326; /note="Proton donor"; /evidence="ECO:0000305|PubMed:10655611, ECO:0000305|PubMed:8407904" |
| Activity Regulation | ACTIVITY REGULATION: Contains three consecutive and one non-consecutive disulfide bonds and shows a strong dependence on DsbC for its full activity (PubMed:15642731). Competitively inhibited by tartaric acid and by sodium fluorid (PubMed:6282821, PubMed:8387749). {ECO:0000269|PubMed:15642731, ECO:0000269|PubMed:6282821, ECO:0000269|PubMed:8387749}. |
| Binding Site | BINDING 38; /note="Phytate"; /evidence="ECO:0000269|PubMed:10655611, ECO:0000305|Ref.18, ECO:0007744|PDB:1DKP, ECO:0007744|PDB:1DKQ"; BINDING 114; /note="Phytate"; /evidence="ECO:0000269|PubMed:10655611, ECO:0000305|Ref.18, ECO:0007744|PDB:1DKP, ECO:0007744|PDB:1DKQ"; BINDING 289; /note="Phytate"; /evidence="ECO:0000269|PubMed:10655611, ECO:0007744|PDB:1DKP, ECO:0007744|PDB:1DKQ" |
| Calcium Binding | |
| catalytic Activity | CATALYTIC ACTIVITY: Reaction=1D-myo-inositol hexakisphosphate + H2O = 1D-myo-inositol 1,2,3,4,5-pentakisphosphate + phosphate; Xref=Rhea:RHEA:68308, ChEBI:CHEBI:15377, ChEBI:CHEBI:43474, ChEBI:CHEBI:58130, ChEBI:CHEBI:177294; Evidence={ECO:0000269|PubMed:10696472, ECO:0000269|PubMed:11035187, ECO:0000269|PubMed:30712472, ECO:0000269|PubMed:8387749, ECO:0000269|Ref.8};PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:68309; Evidence={ECO:0000269|PubMed:10696472, ECO:0000269|PubMed:11035187, ECO:0000269|PubMed:30712472, ECO:0000269|PubMed:8387749, ECO:0000269|Ref.8}; CATALYTIC ACTIVITY: Reaction=1D-myo-inositol 1,2,3,4,5-pentakisphosphate + H2O = 1D-myo-inositol 2,3,4,5-tetrakisphosphate + phosphate; Xref=Rhea:RHEA:68312, ChEBI:CHEBI:15377, ChEBI:CHEBI:43474, ChEBI:CHEBI:177294, ChEBI:CHEBI:177295; Evidence={ECO:0000269|PubMed:11035187, ECO:0000269|PubMed:8387749};PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:68313; Evidence={ECO:0000269|PubMed:11035187, ECO:0000269|PubMed:8387749}; CATALYTIC ACTIVITY: Reaction=1D-myo-inositol 2,3,4,5-tetrakisphosphate + H2O = 1D-myo-inositol 2,4,5-triphosphate + phosphate; Xref=Rhea:RHEA:68316, ChEBI:CHEBI:15377, ChEBI:CHEBI:43474, ChEBI:CHEBI:177295, ChEBI:CHEBI:177296; Evidence={ECO:0000269|PubMed:11035187, ECO:0000269|PubMed:8387749};PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:68317; Evidence={ECO:0000269|PubMed:11035187, ECO:0000269|PubMed:8387749}; CATALYTIC ACTIVITY: Reaction=1D-myo-inositol 2,4,5-triphosphate + H2O = 1D-myo-inositol 2,5-bisphosphate + phosphate; Xref=Rhea:RHEA:68320, ChEBI:CHEBI:15377, ChEBI:CHEBI:43474, ChEBI:CHEBI:177296, ChEBI:CHEBI:177297; Evidence={ECO:0000269|PubMed:11035187};PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:68321; Evidence={ECO:0000269|PubMed:11035187}; CATALYTIC ACTIVITY: Reaction=1D-myo-inositol 2,5-bisphosphate + H2O = 1D-myo-inositol 2-phosphate + phosphate; Xref=Rhea:RHEA:68324, ChEBI:CHEBI:15377, ChEBI:CHEBI:43474, ChEBI:CHEBI:84142, ChEBI:CHEBI:177297; Evidence={ECO:0000269|PubMed:11035187};PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:68325; Evidence={ECO:0000269|PubMed:11035187}; CATALYTIC ACTIVITY: Reaction=GTP + H2O = GDP + H(+) + phosphate; Xref=Rhea:RHEA:19669, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:37565, ChEBI:CHEBI:43474, ChEBI:CHEBI:58189; Evidence={ECO:0000269|PubMed:6282821};PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:19670; Evidence={ECO:0000269|PubMed:6282821}; |
| DNA Binding | |
| EC Number | 3.1.3.-; 3.6.1.- |
| Enzyme Function | FUNCTION: Catalyzes the hydrolysis of phytate (or myo-inositol hexakisphosphate, an indigestible organic form of phosphorus that is found in many plant tissues) to myo-inositol and inorganic phosphate (PubMed:8387749, Ref.8, PubMed:10696472, PubMed:11035187, PubMed:30712472). Dephosphorylates phytate in a stereospecific way by sequential removal of phosphate groups to produce myo-inositol 2-monophosphate (PubMed:11035187). Also shows phosphoanhydride phosphatase activity and hydrolyzes the distal phosphoryl residues of GTP, the 5'-beta-phosphoryl residue of the regulatory nucleotide ppGpp and tripolyphosphates (PubMed:6282821, PubMed:1429631, PubMed:8407904). Does not split most phosphomonoesters with the exception of the synthetic substrate p-nitrophenyl phosphate (pNPP), 2,3-bisphosphoglycerate and fructose 1,6-bisphosphate (PubMed:6282821, Ref.8, PubMed:1429631, PubMed:8387749, PubMed:8407904, PubMed:10696472). {ECO:0000269|PubMed:10696472, ECO:0000269|PubMed:11035187, ECO:0000269|PubMed:1429631, ECO:0000269|PubMed:30712472, ECO:0000269|PubMed:6282821, ECO:0000269|PubMed:8387749, ECO:0000269|PubMed:8407904, ECO:0000269|Ref.8}. |
| Temperature Dependency | BIOPHYSICOCHEMICAL PROPERTIES: Temperature dependence: Optimum temperature is 55 degrees Celsius for phytase activity (PubMed:8387749). Optimum temperature is 60 degrees Celsius for phytase activity (PubMed:10696472). {ECO:0000269|PubMed:10696472, ECO:0000269|PubMed:8387749}; |
| PH Dependency | BIOPHYSICOCHEMICAL PROPERTIES: pH dependence: Optimum pH is 4.5 for phytase activity (PubMed:8387749, PubMed:10696472). Optimum pH is 2.5 for acid phosphatase activity (PubMed:6282821). Optimum pH is 2.5-3.0 for acid phosphatase activity (PubMed:10696472). {ECO:0000269|PubMed:10696472, ECO:0000269|PubMed:6282821, ECO:0000269|PubMed:8387749}; |
| Pathway | |
| nucleotide Binding | |
| Features | Active site (2); Beta strand (14); Binding site (3); Chain (1); Disulfide bond (4); Helix (19); Mutagenesis (31); Region (2); Sequence conflict (3); Signal peptide (1); Turn (6) |
| Keywords | 3D-structure;Direct protein sequencing;Disulfide bond;Hydrolase;Periplasm;Reference proteome;Signal |
| Interact With | |
| Induction | INDUCTION: Induced by phosphate starvation, anaerobiosis and entry into stationary phase (PubMed:6282821, PubMed:8387749, PubMed:8071219). Regulated by the HTH-type transcriptional regulator AppY (PubMed:8071219). Negatively controlled by the cAMP-cAMP receptor (CAP) complex (PubMed:6282821, PubMed:3038201). {ECO:0000269|PubMed:3038201, ECO:0000269|PubMed:6282821, ECO:0000269|PubMed:8071219, ECO:0000269|PubMed:8387749}. |
| Subcellular Location | SUBCELLULAR LOCATION: Periplasm {ECO:0000269|PubMed:10696472, ECO:0000269|PubMed:1429631, ECO:0000269|PubMed:8387749, ECO:0000269|PubMed:8407904}. |
| Modified Residue | |
| Post Translational Modification | |
| Signal Peptide | SIGNAL 1..22; /evidence="ECO:0000269|PubMed:2168385, ECO:0000269|PubMed:8387749, ECO:0000269|Ref.8" |
| Structure 3D | X-ray crystallography (7) |
| Cross Reference PDB | 1DKL; 1DKM; 1DKN; 1DKO; 1DKP; 1DKQ; 4TSR; |
| Mapped Pubmed ID | 16606699; |
| Motif | |
| Gene Encoded By | |
| Mass | 47,057 |
| Kinetics | BIOPHYSICOCHEMICAL PROPERTIES: Kinetic parameters: KM=0.13 mM for phytate {ECO:0000269|Ref.8}; KM=0.63 mM for phytate {ECO:0000269|PubMed:10696472}; KM=15 uM for D-Ins(1,2,3,4,5)P5 {ECO:0000269|PubMed:11035187}; KM=0.35 mM for GTP {ECO:0000269|PubMed:6282821}; KM=1.8 mM for ppGpp {ECO:0000269|PubMed:6282821}; KM=0.15 mM for tripolyphosphate {ECO:0000269|PubMed:1429631}; KM=6.5 mM for GDP {ECO:0000269|PubMed:6282821}; KM=20 mM for ATP {ECO:0000269|PubMed:6282821}; KM=2.77 mM for pNPP {ECO:0000269|PubMed:6282821}; KM=2.6 mM for pNPP {ECO:0000269|Ref.8}; KM=2.8 mM for pNPP {ECO:0000269|PubMed:1429631}; KM=5 mM for 2,3-bisphosphoglycerate {ECO:0000269|PubMed:6282821}; KM=5 mM for fructose 1,6-bisphosphate {ECO:0000269|PubMed:6282821}; KM=3.0 mM for fructose 1,6-bisphosphate {ECO:0000269|PubMed:1429631}; Vmax=2326 umol/min/mg enzyme with phytate as substrate {ECO:0000269|PubMed:10696472}; Vmax=9235 umol/min/mg enzyme with D-Ins(1,2,3,4,5)P5 as substrate {ECO:0000269|PubMed:11035187}; Vmax=22 nmol/min/mg enzyme with GTP as substrate {ECO:0000269|PubMed:6282821}; Vmax=40 nmol/min/mg enzyme with ppGpp as substrate {ECO:0000269|PubMed:6282821}; Vmax=440 umol/min/mg enzyme with tripolyphosphate as substrate {ECO:0000269|PubMed:1429631}; Vmax=5 nmol/min/mg enzyme with GDP as substrate {ECO:0000269|PubMed:6282821}; Vmax=62 nmol/min/mg enzyme with ATP as substrate {ECO:0000269|PubMed:6282821}; Vmax=208 nmol/min/mg enzyme with pNPP as substrate {ECO:0000269|PubMed:6282821}; Vmax=530 umol/min/mg enzyme with pNPP as substrate {ECO:0000269|PubMed:1429631}; Vmax=625 nmol/min/mg enzyme with 2,3-bisphosphoglycerate as substrate {ECO:0000269|PubMed:6282821}; Vmax=384 nmol/min/mg enzyme with fructose 1,6-bisphosphate as substrate {ECO:0000269|PubMed:6282821}; Vmax=764 umol/min/mg enzyme with fructose 1,6-bisphosphate as substrate {ECO:0000269|PubMed:1429631}; |
| Metal Binding | |
| Rhea ID | RHEA:68308; RHEA:68309; RHEA:68312; RHEA:68313; RHEA:68316; RHEA:68317; RHEA:68320; RHEA:68321; RHEA:68324; RHEA:68325; RHEA:19669; RHEA:19670 |
| Cross Reference Brenda | 3.1.3.26; |