| IED ID | IndEnz0010001356 |
| Enzyme Type ID | esterase001356 |
| Protein Name |
Leaf-branch compost cutinase LC-cutinase LCC EC 3.1.1.74 PET-digesting enzyme Poly ethylene terephthalate hydrolase PET hydrolase PETase EC 3.1.1.101 |
| Gene Name | |
| Organism | Unknown prokaryotic organism |
| Taxonomic Lineage | cellular organisms Bacteria environmental samples Unknown prokaryotic organism |
| Enzyme Sequence | MDGVLWRVRTAALMAALLALAAWALVWASPSVEAQSNPYQRGPNPTRSALTADGPFSVATYTVSRLSVSGFGGGVIYYPTGTSLTFGGIAMSPGYTADASSLAWLGRRLASHGFVVLVINTNSRFDYPDSRASQLSAALNYLRTSSPSAVRARLDANRLAVAGHSMGGGGTLRIAEQNPSLKAAVPLTPWHTDKTFNTSVPVLIVGAEADTVAPVSQHAIPFYQNLPSTTPKVYVELDNASHFAPNSNNAAISVYTISWMKLWVDNDTRYRQFLCNVNDPALSDFRTNNRHCQ |
| Enzyme Length | 293 |
| Uniprot Accession Number | G9BY57 |
| Absorption | |
| Active Site | ACT_SITE 165; /note="Nucleophile"; /evidence="ECO:0000305|PubMed:24593046, ECO:0000305|PubMed:32269349"; ACT_SITE 210; /note="Charge relay system"; /evidence="ECO:0000305|PubMed:24593046, ECO:0000305|PubMed:32269349"; ACT_SITE 242; /note="Charge relay system"; /evidence="ECO:0000305|PubMed:24593046, ECO:0000305|PubMed:32269349" |
| Activity Regulation | ACTIVITY REGULATION: Is inhibited in vitro by diethyl pNP-phosphate (E600), but not by EDTA. {ECO:0000269|PubMed:22194294}. |
| Binding Site | BINDING 95; /note=Poly(ethylene terephthalate); /evidence=ECO:0000250|UniProtKB:A0A0K8P6T7; BINDING 166; /note=Poly(ethylene terephthalate); /evidence=ECO:0000250|UniProtKB:A0A0K8P6T7; BINDING 190; /note=Poly(ethylene terephthalate); /evidence=ECO:0000250|UniProtKB:A0A0K8P6T7 |
| Calcium Binding | |
| catalytic Activity | CATALYTIC ACTIVITY: Reaction=Cutin + H(2)O = cutin monomers.; EC=3.1.1.74; Evidence={ECO:0000269|PubMed:22194294}; 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:22194294, ECO:0000269|PubMed:32269349}; CATALYTIC ACTIVITY: Reaction=4-[(2-hydroxyethoxy)carbonyl]benzoate + H2O = ethylene glycol + H(+) + terephthalate; Xref=Rhea:RHEA:49532, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:30043, ChEBI:CHEBI:30742, ChEBI:CHEBI:131704; Evidence={ECO:0000269|PubMed:22194294, ECO:0000269|PubMed:32269349}; 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:22194294}; 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:22194294, ECO:0000269|PubMed:24593046}; 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:22194294}; 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:22194294}; |
| DNA Binding | |
| EC Number | 3.1.1.74; 3.1.1.101 |
| Enzyme Function | FUNCTION: Catalyzes the hydrolysis of cutin, a polyester that forms the structure of plant cuticle (PubMed:22194294). Shows esterase activity towards p-nitrophenol-linked aliphatic esters (pNP-aliphatic esters), with a preference for short-chain substrates (C4 substrate at most) (PubMed:22194294, PubMed:24593046). Cannot hydrolyze olive oil (PubMed:22194294). Is also able to degrade poly(ethylene terephthalate), the most abundant polyester plastic in the world (PubMed:22194294, PubMed:32269349). Can also depolymerize poly(epsilon-caprolactone) (PCL), a synthetic aliphatic biodegradable polyester (PubMed:22194294). {ECO:0000269|PubMed:22194294, ECO:0000269|PubMed:24593046, ECO:0000269|PubMed:32269349}. |
| Temperature Dependency | BIOPHYSICOCHEMICAL PROPERTIES: Temperature dependence: Optimum temperature is 50 degrees Celsius with pNP-butanoate as substrate (PubMed:22194294, PubMed:24593046). Optimum temperature using PET as substrate is superior to 70 degrees Celsius (PubMed:22194294). Shows 70% of the maximal activity at 30 and 70 degrees Celsius with pNP-butyrate as substrate. Has half-lives of 5 hours at 50 degrees Celsius, 80 minutes at 60 degrees Celsius, 40 minutes at 70 degrees Celsius and 7 minutes at 80 degrees Celsius (PubMed:22194294). Is thermostable, with a determined melting temperature (Tm) of 84.7 degrees Celsius (PubMed:32269349). {ECO:0000269|PubMed:22194294, ECO:0000269|PubMed:24593046, ECO:0000269|PubMed:32269349}; |
| PH Dependency | BIOPHYSICOCHEMICAL PROPERTIES: pH dependence: Optimum pH is 8.5 with pNP-butyrate as substrate. Shows 70% of the maximal activity at pH 7.0 and pH 9.5. {ECO:0000269|PubMed:22194294}; |
| Pathway | |
| nucleotide Binding | |
| Features | Active site (3); Beta strand (11); Binding site (3); Chain (1); Disulfide bond (1); Helix (12); Mutagenesis (2); Signal peptide (1); Turn (1) |
| Keywords | 3D-structure;Disulfide bond;Hydrolase;Secreted;Serine esterase;Signal |
| Interact With | |
| Induction | |
| Subcellular Location | SUBCELLULAR LOCATION: Secreted {ECO:0000305|PubMed:22194294}. |
| Modified Residue | |
| Post Translational Modification | PTM: The disulfide bond between Cys-275 and Cys-292 contributes not only to the thermodynamic stability but also to the kinetic stability of the enzyme. {ECO:0000269|PubMed:24593046}. |
| Signal Peptide | SIGNAL 1..34; /evidence=ECO:0000255 |
| Structure 3D | X-ray crystallography (3) |
| Cross Reference PDB | 4EB0; 6THS; 6THT; |
| Mapped Pubmed ID | - |
| Motif | |
| Gene Encoded By | |
| Mass | 31,496 |
| Kinetics | BIOPHYSICOCHEMICAL PROPERTIES: Kinetic parameters: KM=0.22 mM for pNP-butanoate (at 30 degrees Celsius and pH 8.0) {ECO:0000269|PubMed:24593046}; KM=0.21 mM for pNP-butanoate (at 50 degrees Celsius and pH 8.0) {ECO:0000269|PubMed:24593046}; KM=0.24 mM for pNP-butanoate (at 70 degrees Celsius and pH 8.0) {ECO:0000269|PubMed:24593046}; Note=kcat is 232 sec(-1) with pNP-butanoate as substrate (at 30 degrees Celsius and pH 8.0). kcat is 343 sec(-1) with pNP-butanoate as substrate (at 50 degrees Celsius and pH 8.0). kcat is 318 sec(-1) with pNP-butanoate as substrate (at 70 degrees Celsius and pH 8.0). {ECO:0000269|PubMed:24593046}; |
| Metal Binding | |
| Rhea ID | RHEA:49528; RHEA:49532; RHEA:12957; RHEA:47348; RHEA:47352; RHEA:47356 |
| Cross Reference Brenda |