| IED ID | IndEnz0010001064 |
| Enzyme Type ID | esterase001064 |
| Protein Name |
Poly ethylene terephthalate hydrolase PET hydrolase PETase EC 3.1.1.101 PET-digesting enzyme |
| Gene Name | ISF6_4831 |
| Organism | Ideonella sakaiensis (strain NBRC 110686 / TISTR 2288 / 201-F6) |
| Taxonomic Lineage | cellular organisms Bacteria Proteobacteria Betaproteobacteria Burkholderiales Burkholderiales genera incertae sedis Ideonella Ideonella sakaiensis (strain NBRC 110686 / TISTR 2288 / 201-F6) |
| Enzyme Sequence | MNFPRASRLMQAAVLGGLMAVSAAATAQTNPYARGPNPTAASLEASAGPFTVRSFTVSRPSGYGAGTVYYPTNAGGTVGAIAIVPGYTARQSSIKWWGPRLASHGFVVITIDTNSTLDQPSSRSSQQMAALRQVASLNGTSSSPIYGKVDTARMGVMGWSMGGGGSLISAANNPSLKAAAPQAPWDSSTNFSSVTVPTLIFACENDSIAPVNSSALPIYDSMSRNAKQFLEINGGSHSCANSGNSNQALIGKKGVAWMKRFMDNDTRYSTFACENPNSTRVSDFRTANCS |
| Enzyme Length | 290 |
| Uniprot Accession Number | A0A0K8P6T7 |
| Absorption | |
| Active Site | ACT_SITE 160; /note="Nucleophile"; /evidence="ECO:0000305|PubMed:29235460, ECO:0000305|PubMed:29374183, ECO:0000305|PubMed:29603535, ECO:0000305|PubMed:29666242"; ACT_SITE 206; /note="Charge relay system"; /evidence="ECO:0000305|PubMed:29235460, ECO:0000305|PubMed:29374183, ECO:0000305|PubMed:29603535, ECO:0000305|PubMed:29666242"; ACT_SITE 237; /note="Charge relay system"; /evidence="ECO:0000305|PubMed:29235460, ECO:0000305|PubMed:29374183, ECO:0000305|PubMed:29603535, ECO:0000305|PubMed:29666242" |
| Activity Regulation | ACTIVITY REGULATION: Salts and glycerol enhance the enzymatic activity in vitro towards pNP-esters, while detergents and organic solvents reduce the enzymatic activity. {ECO:0000269|PubMed:30502092}. |
| Binding Site | BINDING 87; /note="Poly(ethylene terephthalate)"; /evidence="ECO:0000269|PubMed:29235460"; BINDING 161; /note="Poly(ethylene terephthalate)"; /evidence="ECO:0000269|PubMed:29235460"; BINDING 185; /note="Poly(ethylene terephthalate)"; /evidence="ECO:0000305|PubMed:29235460, ECO:0000305|PubMed:29666242" |
| Calcium Binding | |
| catalytic Activity | CATALYTIC ACTIVITY: Reaction=(ethylene terephthalate)(n) + H2O = (ethylene terephthalate)(n-1) + 4-[(2-hydroxyethoxy)carbonyl]benzoate + H(+); Xref=Rhea:RHEA:49528, Rhea:RHEA-COMP:12420, Rhea:RHEA-COMP:12421, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:131701, ChEBI:CHEBI:131704; EC=3.1.1.101; Evidence={ECO:0000269|PubMed:26965627, ECO:0000269|PubMed:29235460, ECO:0000269|PubMed:29374183, ECO:0000269|PubMed:29603535, ECO:0000269|PubMed:29666242, ECO:0000269|PubMed:32269349};PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:49529; Evidence={ECO:0000269|PubMed:26965627}; CATALYTIC ACTIVITY: Reaction=(2,5-ethylene furandicarboxylate)(n) + 2 H2O = (2,5-ethylene furandicarboxylate)(n-1) + 2,5-dicarboxyfuran + ethylene glycol + 2 H(+); Xref=Rhea:RHEA:42648, Rhea:RHEA-COMP:14671, Rhea:RHEA-COMP:14672, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:30742, ChEBI:CHEBI:83389, ChEBI:CHEBI:140646; Evidence={ECO:0000269|PubMed:29666242}; CATALYTIC ACTIVITY: Reaction=an acetyl ester + H2O = acetate + an aliphatic alcohol + H(+); Xref=Rhea:RHEA:12957, ChEBI:CHEBI:2571, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:30089, ChEBI:CHEBI:47622; Evidence={ECO:0000269|PubMed:26965627, ECO:0000269|PubMed:30502092}; CATALYTIC ACTIVITY: Reaction=a butanoate ester + H2O = an aliphatic alcohol + butanoate + H(+); Xref=Rhea:RHEA:47348, ChEBI:CHEBI:2571, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:17968, ChEBI:CHEBI:50477; Evidence={ECO:0000269|PubMed:26965627, ECO:0000269|PubMed:30502092}; CATALYTIC ACTIVITY: Reaction=a hexanoate ester + H2O = an aliphatic alcohol + H(+) + hexanoate; Xref=Rhea:RHEA:47352, ChEBI:CHEBI:2571, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:17120, ChEBI:CHEBI:87656; Evidence={ECO:0000269|PubMed:26965627, ECO:0000269|PubMed:30502092}; CATALYTIC ACTIVITY: Reaction=an octanoate ester + H2O = an aliphatic alcohol + H(+) + octanoate; Xref=Rhea:RHEA:47356, ChEBI:CHEBI:2571, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:25646, ChEBI:CHEBI:87657; Evidence={ECO:0000269|PubMed:26965627, ECO:0000269|PubMed:30502092}; CATALYTIC ACTIVITY: Reaction=a dodecanoate ester + H2O = an aliphatic alcohol + dodecanoate + H(+); Xref=Rhea:RHEA:47364, ChEBI:CHEBI:2571, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:18262, ChEBI:CHEBI:87659; Evidence={ECO:0000269|PubMed:30502092}; |
| DNA Binding | |
| EC Number | 3.1.1.101 |
| Enzyme Function | FUNCTION: Involved in the degradation and assimilation of the plastic poly(ethylene terephthalate) (PET), which allows I.sakaiensis to use PET as its major energy and carbon source for growth. Likely acts synergistically with MHETase to depolymerize PET (PubMed:26965627). Catalyzes the hydrolysis of PET to produce mono(2-hydroxyethyl) terephthalate (MHET) as the major product (PubMed:26965627, PubMed:32269349, PubMed:29666242, PubMed:29603535, PubMed:29374183, PubMed:29235460). Also depolymerizes another semiaromatic polyester, poly(ethylene-2,5-furandicarboxylate) (PEF), which is an emerging, bioderived PET replacement with improved gas barrier properties (PubMed:29666242). In contrast, PETase does not degrade aliphatic polyesters such as polylactic acid (PLA) and polybutylene succinate (PBS) (PubMed:29666242). Is also able to hydrolyze bis(hydroxyethyl) terephthalate (BHET) to yield MHET with no further decomposition, but terephthalate (TPA) can also be observed (PubMed:26965627, PubMed:29603535, PubMed:29374183). Shows esterase activity towards p-nitrophenol-linked aliphatic esters (pNP-aliphatic esters) in vitro (PubMed:26965627, PubMed:30502092). {ECO:0000269|PubMed:26965627, ECO:0000269|PubMed:29235460, ECO:0000269|PubMed:29374183, ECO:0000269|PubMed:29603535, ECO:0000269|PubMed:29666242, ECO:0000269|PubMed:30502092, ECO:0000269|PubMed:32269349}. |
| Temperature Dependency | BIOPHYSICOCHEMICAL PROPERTIES: Temperature dependence: Optimum temperature is 40 degrees Celsius for PET film hydrolysis (PubMed:26965627). Optimum temperature is 30 degrees Celsius for PET (commercial drinking bottle) hydrolysis and BHET hydrolysis (PubMed:29603535). Optimum temperature is 35-45 degrees Celsius for the hydrolysis of pNP-esters. Remains active even at 65 degrees Celsius (about 60% of maximum activity) (PubMed:30502092). {ECO:0000269|PubMed:26965627, ECO:0000269|PubMed:29603535, ECO:0000269|PubMed:30502092}; |
| PH Dependency | BIOPHYSICOCHEMICAL PROPERTIES: pH dependence: Optimum pH is 9 for PET film hydrolysis (PubMed:26965627). Optimum pH is 9 for PET (commercial drinking bottle) hydrolysis. Optimum pH is 6.5-8.0 for BHET hydrolysis (PubMed:29603535). Optimum pH is 8.0 for the hydrolysis of pNP-esters. The enzyme is active at pH 6-10, has an optimal pH range of 7-9 and it is rapidly inactivated below pH 7.0 or above pH 9.0 (PubMed:30502092). {ECO:0000269|PubMed:26965627, ECO:0000269|PubMed:29603535, ECO:0000269|PubMed:30502092}; |
| Pathway | PATHWAY: Xenobiotic degradation. {ECO:0000305|PubMed:26965627}. |
| nucleotide Binding | |
| Features | Active site (3); Beta strand (11); Binding site (3); Chain (1); Disulfide bond (2); Helix (9); Mutagenesis (20); Signal peptide (1); Turn (4) |
| Keywords | 3D-structure;Disulfide bond;Hydrolase;Reference proteome;Secreted;Serine esterase;Signal |
| Interact With | |
| Induction | INDUCTION: Highly up-regulated during growth on PET film. {ECO:0000269|PubMed:26965627}. |
| Subcellular Location | SUBCELLULAR LOCATION: Secreted {ECO:0000305|PubMed:26965627}. |
| Modified Residue | |
| Post Translational Modification | |
| Signal Peptide | SIGNAL 1..27; /evidence=ECO:0000255 |
| Structure 3D | X-ray crystallography (28) |
| Cross Reference PDB | 5XFY; 5XFZ; 5XG0; 5XH2; 5XH3; 5XJH; 5YFE; 5YNS; 6ANE; 6EQD; 6EQE; 6EQF; 6EQG; 6EQH; 6IJ3; 6IJ4; 6IJ5; 6IJ6; 6ILW; 6ILX; 6KUO; 6KUQ; 6KUS; 6KY5; 6QGC; 7CQB; 7CY0; 7OSB; |
| Mapped Pubmed ID | 34587366; |
| Motif | |
| Gene Encoded By | |
| Mass | 30,247 |
| Kinetics | BIOPHYSICOCHEMICAL PROPERTIES: Kinetic parameters: KM=0.431 mM for pNP-acetate {ECO:0000269|PubMed:30502092}; KM=0.315 mM for pNP-butanoate {ECO:0000269|PubMed:30502092}; KM=0.053 mM for pNP-hexanoate {ECO:0000269|PubMed:30502092}; KM=0.048 mM for pNP-octanoate {ECO:0000269|PubMed:30502092}; KM=2.283 mM for pNP-dodecanoate {ECO:0000269|PubMed:30502092}; Note=kcat is 1590 sec(-1) for the hydrolysis of pNP-acetate. kcat is 1353 sec(-1) for the hydrolysis of pNP-butanoate. kcat is 1345 sec(-1) for the hydrolysis of pNP-hexanoate. kcat is 519 sec(-1) for the hydrolysis of pNP-octanoate. kcat is 1531 sec(-1) for the hydrolysis of pNP-dodecanoate. {ECO:0000269|PubMed:30502092}; |
| Metal Binding | |
| Rhea ID | RHEA:49528; RHEA:49529; RHEA:42648; RHEA:12957; RHEA:47348; RHEA:47352; RHEA:47356; RHEA:47364 |
| Cross Reference Brenda | 3.1.1.101; |