| IED ID | IndEnz0010001404 |
| Enzyme Type ID | esterase001404 |
| Protein Name |
Cutinase cut2 TfCut2 EC 3.1.1.74 Acetylxylan esterase BTA-hydrolase 1 Poly ethylene terephthalate hydrolase PET hydrolase PETase EC 3.1.1.101 |
| Gene Name | cut2 bta1 cut1 TfH |
| Organism | Thermobifida fusca (Thermomonospora fusca) |
| Taxonomic Lineage | cellular organisms Bacteria Terrabacteria group Actinobacteria Actinomycetia (high G+C Gram-positive bacteria) Streptosporangiales Nocardiopsaceae Thermobifida Thermobifida fusca (Thermomonospora fusca) |
| Enzyme Sequence | MAVMTPRRERSSLLSRALQVTAAAATALVTAVSLAAPAHAANPYERGPNPTDALLEASSGPFSVSEENVSRLSASGFGGGTIYYPRENNTYGAVAISPGYTGTEASIAWLGERIASHGFVVITIDTITTLDQPDSRAEQLNAALNHMINRASSTVRSRIDSSRLAVMGHSMGGGGTLRLASQRPDLKAAIPLTPWHLNKNWSSVTVPTLIIGADLDTIAPVATHAKPFYNSLPSSISKAYLELDGATHFAPNIPNKIIGKYSVAWLKRFVDNDTRYTQFLCPGPRDGLFGEVEEYRSTCPF |
| Enzyme Length | 301 |
| Uniprot Accession Number | Q6A0I4 |
| Absorption | |
| Active Site | ACT_SITE 170; /note="Nucleophile"; /evidence="ECO:0000269|PubMed:24728714, ECO:0007744|PDB:4CG2"; ACT_SITE 216; /note="Charge relay system"; /evidence="ECO:0000250|UniProtKB:A0A0K8P6T7"; ACT_SITE 248; /note="Charge relay system"; /evidence="ECO:0000250|UniProtKB:A0A0K8P6T7" |
| Activity Regulation | ACTIVITY REGULATION: Activated by magnesium ions (PubMed:25545638). Activated by calcium ions (PubMed:25545638). Inhibited by the serine hydrolase inhibitor phenylmethanesulfonyl fluoride (PMSF) (Probable). {ECO:0000269|PubMed:25545638, ECO:0000305|PubMed:24728714}. |
| Binding Site | BINDING 100; /note="Poly(ethylene terephthalate)"; /evidence="ECO:0000250|UniProtKB:A0A0K8P6T7, ECO:0000305|PubMed:24728714"; BINDING 171; /note="Poly(ethylene terephthalate)"; /evidence="ECO:0000250|UniProtKB:A0A0K8P6T7, ECO:0000305|PubMed:24728714"; BINDING 195; /note="Poly(ethylene terephthalate)"; /evidence="ECO:0000250|UniProtKB:A0A0K8P6T7" |
| Calcium Binding | |
| catalytic Activity | CATALYTIC ACTIVITY: Reaction=H2O + pentanoate ester = an aliphatic alcohol + H(+) + pentanoate; Xref=Rhea:RHEA:48436, ChEBI:CHEBI:2571, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:31011, ChEBI:CHEBI:50871; Evidence={ECO:0000269|PubMed:23604968};PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:48437; Evidence={ECO:0000269|PubMed:23604968}; 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:23604968}; CATALYTIC ACTIVITY: Reaction=decanoate ester + H2O = an aliphatic alcohol + decanoate + H(+); Xref=Rhea:RHEA:47360, ChEBI:CHEBI:2571, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:27689, ChEBI:CHEBI:87658; Evidence={ECO:0000269|PubMed:23604968};PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:47361; Evidence={ECO:0000269|PubMed:23604968}; 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:23604968};PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:47365; Evidence={ECO:0000269|PubMed:23604968}; CATALYTIC ACTIVITY: Reaction=a tetradecanoate ester + H2O = an aliphatic alcohol + H(+) + tetradecanoate; Xref=Rhea:RHEA:47388, ChEBI:CHEBI:2571, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:30807, ChEBI:CHEBI:87691; Evidence={ECO:0000269|PubMed:23604968};PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:47389; Evidence={ECO:0000269|PubMed:23604968}; CATALYTIC ACTIVITY: Reaction=H2O + hexadecanoate ester = an aliphatic alcohol + H(+) + hexadecanoate; Xref=Rhea:RHEA:47392, ChEBI:CHEBI:2571, ChEBI:CHEBI:7896, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:25835; Evidence={ECO:0000269|PubMed:23604968};PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:47393; Evidence={ECO:0000269|PubMed:23604968}; CATALYTIC ACTIVITY: Reaction=Cutin + H(2)O = cutin monomers.; EC=3.1.1.74; Evidence={ECO:0000305|PubMed:23604968}; 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:23604968, ECO:0000269|PubMed:24728714, ECO:0000269|Ref.4};PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:12958; Evidence={ECO:0000269|PubMed:23604968, ECO:0000269|PubMed:24728714, ECO:0000269|Ref.4}; 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:25545638, ECO:0000269|PubMed:31690819, ECO:0000269|PubMed:32269349, ECO:0000269|Ref.4};PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:49529; Evidence={ECO:0000269|PubMed:25545638, ECO:0000269|PubMed:31690819, ECO:0000269|PubMed:32269349, ECO:0000269|Ref.4}; 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:15638529, ECO:0000269|PubMed:23604968, ECO:0000269|PubMed:24728714, ECO:0000269|PubMed:25545638, ECO:0000269|PubMed:31690819, ECO:0000269|Ref.4};PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:47349; Evidence={ECO:0000269|PubMed:15638529, ECO:0000269|PubMed:23604968, ECO:0000269|PubMed:24728714, ECO:0000269|PubMed:25545638, ECO:0000269|PubMed:31690819, ECO:0000269|Ref.4}; |
| 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 (Ref.4, PubMed:31690819, PubMed:24728714, PubMed:23604968). Shows esterase activity towards p-nitrophenol-linked aliphatic esters (pNP-aliphatic esters) (Ref.4, PubMed:31690819, PubMed:24728714, PubMed:23604968, PubMed:15638529, PubMed:25545638). Also hydrolyzes the triglycerides triacetin, tributyrin, tricaprin, and trilaurin, with a preference for short-chain substrates (PubMed:15638529). Hydrolyzes the hemicellulose xylan (PubMed:20816933). Capable of degrading the plastic poly(ethylene terephthalate) (PET), the most abundant polyester plastic in the world (Ref.4, PubMed:25545638, PubMed:31690819, PubMed:32269349). Can also depolymerize poly(epsilon-caprolactone) (PCL), a synthetic aliphatic biodegradable polyester (PubMed:15638529). Hydrolyzes polyoxyethylenesorbate esters with a preference for shorter chain lengths (PubMed:20816933). {ECO:0000269|PubMed:15638529, ECO:0000269|PubMed:20816933, ECO:0000269|PubMed:23604968, ECO:0000269|PubMed:24728714, ECO:0000269|PubMed:25545638, ECO:0000269|PubMed:31690819, ECO:0000269|PubMed:32269349, ECO:0000269|Ref.4}. |
| Temperature Dependency | BIOPHYSICOCHEMICAL PROPERTIES: Temperature dependence: Optimum temperature is 25-50 degrees Celsius (PubMed:24728714). Optimum temperature is 55 degrees Celsius (PubMed:23604968). Optimum temperature is 60 degrees Celsius (PubMed:20816933). Optimum temperature is 65 degrees Celsius (PubMed:15638529, PubMed:31690819, PubMed:32269349). {ECO:0000269|PubMed:15638529, ECO:0000269|PubMed:20816933, ECO:0000269|PubMed:23604968, ECO:0000269|PubMed:24728714, ECO:0000269|PubMed:31690819, ECO:0000269|PubMed:32269349}; |
| PH Dependency | BIOPHYSICOCHEMICAL PROPERTIES: pH dependence: Optimum pH is 8 (PubMed:23604968). Optimum pH is 6.0-6.5 (PubMed:15638529). Optimum pH is 7.5 (PubMed:20816933). {ECO:0000269|PubMed:15638529, ECO:0000269|PubMed:20816933, ECO:0000269|PubMed:23604968}; |
| Pathway | |
| nucleotide Binding | |
| Features | Active site (3); Binding site (3); Chain (1); Disulfide bond (1); Mutagenesis (7); Sequence conflict (11); Signal peptide (1) |
| Keywords | 3D-structure;Disulfide bond;Hydrolase;Periplasm;Secreted;Serine esterase;Signal |
| Interact With | |
| Induction | |
| Subcellular Location | SUBCELLULAR LOCATION: Secreted {ECO:0000269|PubMed:23604968}. Periplasm {ECO:0000269|PubMed:23604968}. |
| Modified Residue | |
| Post Translational Modification | |
| Signal Peptide | SIGNAL 1..40; /evidence=ECO:0000255 |
| Structure 3D | X-ray crystallography (4) |
| Cross Reference PDB | 4CG1; 4CG2; 4CG3; 5ZOA; |
| Mapped Pubmed ID | - |
| Motif | |
| Gene Encoded By | |
| Mass | 32,218 |
| Kinetics | BIOPHYSICOCHEMICAL PROPERTIES: Kinetic parameters: KM=167 uM for pNP-acetate (at 25 degrees Celsius and pH 7) {ECO:0000269|Ref.4}; KM=89 uM for pNP-butanoate (at 50 degrees Celsius and pH 8) {ECO:0000269|PubMed:23604968}; KM=2100 uM for pNP-butanoate (at 25 degrees Celsius and pH 7) {ECO:0000269|Ref.4}; Note=kcat is 39.5 sec(-1) with pNP-acetate as substrate (at 25 degrees Celsius and pH 7) (Ref.4). kcat is 253 sec(-1) with pNP-butanoate as substrate (at 50 degrees Celsius and pH 8) (PubMed:23604968). kcat is 30.9 sec(-1) with pNP-butanoate as substrate (at 25 degrees Celsius and pH 7) (Ref.4). {ECO:0000269|PubMed:23604968, ECO:0000269|Ref.4}; |
| Metal Binding | |
| Rhea ID | RHEA:48436; RHEA:48437; RHEA:47356; RHEA:47360; RHEA:47361; RHEA:47364; RHEA:47365; RHEA:47388; RHEA:47389; RHEA:47392; RHEA:47393; RHEA:12957; RHEA:12958; RHEA:49528; RHEA:49529; RHEA:47348; RHEA:47349 |
| Cross Reference Brenda | 3.1.1.74; |