PETH2_THEFY
ID PETH2_THEFY Reviewed; 301 AA.
AC Q47RJ6;
DT 25-MAY-2022, integrated into UniProtKB/Swiss-Prot.
DT 13-SEP-2005, sequence version 1.
DT 03-AUG-2022, entry version 104.
DE RecName: Full=Cutinase;
DE EC=3.1.1.74 {ECO:0000269|PubMed:18658138, ECO:0000269|PubMed:20729325};
DE AltName: Full=Poly(ethylene terephthalate) hydrolase {ECO:0000305};
DE Short=PET hydrolase {ECO:0000305};
DE Short=PETase {ECO:0000305};
DE EC=3.1.1.101 {ECO:0000269|PubMed:21751386, ECO:0000269|PubMed:25545638};
DE Flags: Precursor;
GN Name=TfH {ECO:0000303|PubMed:25545638};
GN OrderedLocusNames=Tfu_0883 {ECO:0000312|EMBL:AAZ54921.1};
OS Thermobifida fusca (strain YX).
OC Bacteria; Actinobacteria; Streptosporangiales; Nocardiopsaceae;
OC Thermobifida.
OX NCBI_TaxID=269800;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=YX;
RX PubMed=17209016; DOI=10.1128/jb.01899-06;
RA Lykidis A., Mavromatis K., Ivanova N., Anderson I., Land M., DiBartolo G.,
RA Martinez M., Lapidus A., Lucas S., Copeland A., Richardson P., Wilson D.B.,
RA Kyrpides N.;
RT "Genome sequence and analysis of the soil cellulolytic actinomycete
RT Thermobifida fusca YX.";
RL J. Bacteriol. 189:2477-2486(2007).
RN [2] {ECO:0000305}
RP FUNCTION, CATALYTIC ACTIVITY, ACTIVITY REGULATION, BIOPHYSICOCHEMICAL
RP PROPERTIES, AND BIOTECHNOLOGY.
RX PubMed=18658138; DOI=10.1074/jbc.m800848200;
RA Chen S., Tong X., Woodard R.W., Du G., Wu J., Chen J.;
RT "Identification and characterization of bacterial cutinase.";
RL J. Biol. Chem. 283:25854-25862(2008).
RN [3] {ECO:0000305}
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, AND
RP BIOTECHNOLOGY.
RX PubMed=20729325; DOI=10.1128/aem.00896-10;
RA Zhang Y., Chen S., Xu M., Cavaco-Paulo A., Cavoco-Paulo A., Wu J., Chen J.;
RT "Characterization of Thermobifida fusca cutinase-carbohydrate-binding
RT module fusion proteins and their potential application in bioscouring.";
RL Appl. Environ. Microbiol. 76:6870-6876(2010).
RN [4] {ECO:0000305}
RP FUNCTION, AND CATALYTIC ACTIVITY.
RX PubMed=21594592; DOI=10.1007/s12010-011-9286-z;
RA Chen S., Liu Z., Chen J., Wu J.;
RT "Study on improvement of extracellular production of recombinant
RT Thermobifida fusca cutinase by Escherichia coli.";
RL Appl. Biochem. Biotechnol. 165:666-675(2011).
RN [5] {ECO:0000305}
RP FUNCTION, CATALYTIC ACTIVITY, BIOTECHNOLOGY, AND MUTAGENESIS OF THR-101;
RP GLN-132 AND ILE-218.
RX PubMed=21751386; DOI=10.1002/biot.201000391;
RA Silva C., Da S., Silva N., Matama T., Araujo R., Martins M., Chen S.,
RA Chen J., Wu J., Casal M., Cavaco-Paulo A.;
RT "Engineered Thermobifida fusca cutinase with increased activity on
RT polyester substrates.";
RL Biotechnol. J. 6:1230-1239(2011).
RN [6] {ECO:0000305}
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, AND
RP MUTAGENESIS OF SER-170.
RX PubMed=23603671; DOI=10.1128/aem.00239-13;
RA Su L., Woodard R.W., Chen J., Wu J.;
RT "Extracellular location of Thermobifida fusca cutinase expressed in
RT Escherichia coli BL21(DE3) without mediation of a signal peptide.";
RL Appl. Environ. Microbiol. 79:4192-4198(2013).
RN [7] {ECO:0000305}
RP FUNCTION, CATALYTIC ACTIVITY, ACTIVITY REGULATION, AND BIOTECHNOLOGY.
RX PubMed=25545638; DOI=10.1002/biot.201400620;
RA Then J., Wei R., Oeser T., Barth M., Belisario-Ferrari M.R., Schmidt J.,
RA Zimmermann W.;
RT "Ca2+ and Mg2+ binding site engineering increases the degradation of
RT polyethylene terephthalate films by polyester hydrolases from Thermobifida
RT fusca.";
RL Biotechnol. J. 10:592-598(2015).
RN [8] {ECO:0000305}
RP BIOTECHNOLOGY.
RX PubMed=27041308; DOI=10.1016/j.foodchem.2016.03.051;
RA Su L., Hong R., Guo X., Wu J., Xia Y.;
RT "Short-chain aliphatic ester synthesis using Thermobifida fusca cutinase.";
RL Food Chem. 206:131-136(2016).
CC -!- FUNCTION: Catalyzes the hydrolysis of cutin, a polyester that forms the
CC structure of plant cuticle (PubMed:18658138, PubMed:20729325). Shows
CC esterase activity towards p-nitrophenol-linked aliphatic esters (pNP-
CC aliphatic esters) (PubMed:18658138, PubMed:20729325, PubMed:21594592,
CC PubMed:21751386, PubMed:23603671, PubMed:25545638). Also hydrolyzes the
CC triglyceride triolein (PubMed:18658138, PubMed:20729325,
CC PubMed:21751386) (Probable). Capable of degrading the plastic
CC poly(ethylene terephthalate) (PET), the most abundant polyester plastic
CC in the world (PubMed:21751386, PubMed:25545638).
CC {ECO:0000269|PubMed:18658138, ECO:0000269|PubMed:20729325,
CC ECO:0000269|PubMed:21594592, ECO:0000269|PubMed:21751386,
CC ECO:0000269|PubMed:23603671, ECO:0000269|PubMed:25545638,
CC ECO:0000305|PubMed:23603671}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=a butanoate ester + H2O = an aliphatic alcohol + butanoate +
CC H(+); Xref=Rhea:RHEA:47348, ChEBI:CHEBI:2571, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:17968, ChEBI:CHEBI:50477;
CC Evidence={ECO:0000269|PubMed:18658138, ECO:0000269|PubMed:20729325,
CC ECO:0000269|PubMed:21594592, ECO:0000269|PubMed:21751386,
CC ECO:0000269|PubMed:23603671, ECO:0000269|PubMed:25545638};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:47349;
CC Evidence={ECO:0000269|PubMed:18658138, ECO:0000269|PubMed:20729325,
CC ECO:0000269|PubMed:21751386, ECO:0000269|PubMed:25545638};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(ethylene terephthalate)(n) + H2O = (ethylene
CC terephthalate)(n-1) + 4-[(2-hydroxyethoxy)carbonyl]benzoate + H(+);
CC Xref=Rhea:RHEA:49528, Rhea:RHEA-COMP:12420, Rhea:RHEA-COMP:12421,
CC ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:131701,
CC ChEBI:CHEBI:131704; EC=3.1.1.101;
CC Evidence={ECO:0000269|PubMed:21751386, ECO:0000269|PubMed:25545638};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:49529;
CC Evidence={ECO:0000269|PubMed:21751386, ECO:0000269|PubMed:25545638};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=cutin + H2O = cutin monomers.; EC=3.1.1.74;
CC Evidence={ECO:0000269|PubMed:18658138, ECO:0000269|PubMed:20729325};
CC -!- ACTIVITY REGULATION: Activated by magnesium ions (PubMed:25545638).
CC Activated by calcium ions (PubMed:25545638). Inhibited by the serine
CC hydrolase inhibitor phenylmethanesulfonyl fluoride (PMSF)
CC (PubMed:18658138). {ECO:0000269|PubMed:18658138,
CC ECO:0000269|PubMed:25545638}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=640 uM for pNP-butanoate {ECO:0000269|PubMed:20729325};
CC KM=312 uM for pNP-butanoate (at 20 degrees Celsius and pH 8)
CC {ECO:0000269|PubMed:23603671};
CC Note=kcat is 220 sec(-1) with pNP-butanoate as substrate
CC (PubMed:20729325). kcat is 304 sec(-1) with pNP-butanoate as
CC substrate (at 20 degrees Celsius and pH 8) (PubMed:23603671).
CC {ECO:0000269|PubMed:20729325, ECO:0000269|PubMed:23603671};
CC pH dependence:
CC Optimum pH is 8. {ECO:0000269|PubMed:18658138,
CC ECO:0000269|PubMed:20729325, ECO:0000269|PubMed:23603671};
CC Temperature dependence:
CC Optimum temperature is 60 degrees Celsius.
CC {ECO:0000269|PubMed:18658138, ECO:0000269|PubMed:20729325,
CC ECO:0000269|PubMed:23603671};
CC -!- SUBCELLULAR LOCATION: Secreted {ECO:0000250|UniProtKB:G8GER6}.
CC Periplasm {ECO:0000250|UniProtKB:G8GER6}.
CC -!- BIOTECHNOLOGY: May have promising applications in chemical and textile
CC industries as it is capable of hydrolyzing a variety of substrates
CC including soluble esters and insoluble triglycerides (PubMed:18658138,
CC PubMed:20729325). Can hydrolyze and thus modulate the surface
CC properties of natural fibers such as cotton (PubMed:20729325). Has
CC potential for application in biological recycling of plastic waste
CC products (PubMed:21751386, PubMed:25545638). As the hydrolysis reaction
CC is reversible, may also have applications in the food industry for
CC synthesizing short-chain aliphatic esters, odor chemicals used as food
CC flavorings (PubMed:27041308). {ECO:0000269|PubMed:18658138,
CC ECO:0000269|PubMed:20729325, ECO:0000269|PubMed:21751386,
CC ECO:0000269|PubMed:25545638, ECO:0000269|PubMed:27041308}.
CC -!- SIMILARITY: Belongs to the AB hydrolase superfamily. {ECO:0000305}.
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DR EMBL; CP000088; AAZ54921.1; -; Genomic_DNA.
DR RefSeq; WP_011291330.1; NC_007333.1.
DR SMR; Q47RJ6; -.
DR STRING; 269800.Tfu_0883; -.
DR ESTHER; thefu-q6a0i4; Polyesterase-lipase-cutinase.
DR EnsemblBacteria; AAZ54921; AAZ54921; Tfu_0883.
DR KEGG; tfu:Tfu_0883; -.
DR eggNOG; COG4188; Bacteria.
DR HOGENOM; CLU_052605_1_0_11; -.
DR OMA; ATHFTPN; -.
DR OrthoDB; 685252at2; -.
DR BRENDA; 3.1.1.3; 6298.
DR BRENDA; 3.1.1.74; 6298.
DR SABIO-RK; Q47RJ6; -.
DR GO; GO:0005576; C:extracellular region; IEA:UniProtKB-SubCell.
DR GO; GO:0042597; C:periplasmic space; IEA:UniProtKB-SubCell.
DR GO; GO:0052689; F:carboxylic ester hydrolase activity; IDA:UniProtKB.
DR GO; GO:0050525; F:cutinase activity; IDA:UniProtKB.
DR GO; GO:0004806; F:triglyceride lipase activity; IEA:UniProtKB-EC.
DR Gene3D; 3.40.50.1820; -; 1.
DR InterPro; IPR029058; AB_hydrolase.
DR InterPro; IPR041127; Chlorophyllase2.
DR Pfam; PF12740; Chlorophyllase2; 1.
DR SUPFAM; SSF53474; SSF53474; 1.
PE 1: Evidence at protein level;
KW Disulfide bond; Hydrolase; Periplasm; Secreted; Serine esterase; Signal.
FT SIGNAL 1..40
FT /evidence="ECO:0000305|PubMed:18658138"
FT CHAIN 41..301
FT /note="Cutinase"
FT /evidence="ECO:0000255"
FT /id="PRO_5004233794"
FT ACT_SITE 170
FT /note="Nucleophile"
FT /evidence="ECO:0000305|PubMed:18658138"
FT ACT_SITE 216
FT /note="Charge relay system"
FT /evidence="ECO:0000250|UniProtKB:A0A0K8P6T7"
FT ACT_SITE 248
FT /note="Charge relay system"
FT /evidence="ECO:0000250|UniProtKB:A0A0K8P6T7"
FT BINDING 100
FT /ligand="poly(ethylene terephthalate)"
FT /ligand_id="ChEBI:CHEBI:131701"
FT /evidence="ECO:0000250|UniProtKB:A0A0K8P6T7"
FT BINDING 171
FT /ligand="poly(ethylene terephthalate)"
FT /ligand_id="ChEBI:CHEBI:131701"
FT /evidence="ECO:0000250|UniProtKB:A0A0K8P6T7"
FT BINDING 195
FT /ligand="poly(ethylene terephthalate)"
FT /ligand_id="ChEBI:CHEBI:131701"
FT /evidence="ECO:0000250|UniProtKB:A0A0K8P6T7"
FT DISULFID 281..299
FT /evidence="ECO:0000250|UniProtKB:A0A0K8P6T7"
FT MUTAGEN 101
FT /note="T->A: Increases activity on poly(ethylene
FT terephthalate) (PET), pNP-butyrate and triolein; when
FT associated with A-132."
FT /evidence="ECO:0000269|PubMed:21751386"
FT MUTAGEN 132
FT /note="Q->A: Increases activity on poly(ethylene
FT terephthalate) (PET), pNP-butyrate and triolein; when
FT associated with A-101."
FT /evidence="ECO:0000269|PubMed:21751386"
FT MUTAGEN 170
FT /note="S->A: Abolishes activity."
FT /evidence="ECO:0000269|PubMed:23603671"
FT MUTAGEN 218
FT /note="I->A: Increases activity on poly(ethylene
FT terephthalate) (PET), pNP-butyrate and triolein."
FT /evidence="ECO:0000269|PubMed:21751386"
SQ SEQUENCE 301 AA; 32218 MW; 4FF805EBE36D888F CRC64;
MAVMTPRRER SSLLSRALQV TAAAATALVT AVSLAAPAHA ANPYERGPNP TDALLEASSG
PFSVSEENVS RLSASGFGGG TIYYPRENNT YGAVAISPGY TGTEASIAWL GERIASHGFV
VITIDTITTL DQPDSRAEQL NAALNHMINR ASSTVRSRID SSRLAVMGHS MGGGGTLRLA
SQRPDLKAAI PLTPWHLNKN WSSVTVPTLI IGADLDTIAP VATHAKPFYN SLPSSISKAY
LELDGATHFA PNIPNKIIGK YSVAWLKRFV DNDTRYTQFL CPGPRDGLFG EVEEYRSTCP
F
