PETH2_THEAE
ID PETH2_THEAE Reviewed; 300 AA.
AC F7IX06;
DT 25-MAY-2022, integrated into UniProtKB/Swiss-Prot.
DT 21-SEP-2011, sequence version 1.
DT 03-AUG-2022, entry version 38.
DE RecName: Full=Cutinase est2 {ECO:0000303|PubMed:22183084};
DE EC=3.1.1.74 {ECO:0000305|PubMed:20393707, ECO:0000305|PubMed:22183084, ECO:0000305|PubMed:25910960, ECO:0000305|PubMed:33387709};
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:0000305|PubMed:20393707, ECO:0000305|PubMed:22183084};
DE AltName: Full=TaCut2 {ECO:0000303|PubMed:30761732};
DE Flags: Precursor;
GN Name=est2 {ECO:0000303|PubMed:22183084};
GN Synonyms=est119 {ECO:0000303|PubMed:20393707};
OS Thermobifida alba (Thermomonospora alba).
OC Bacteria; Actinobacteria; Streptosporangiales; Nocardiopsaceae;
OC Thermobifida.
OX NCBI_TaxID=53522;
RN [1] {ECO:0000312|EMBL:BAK48590.1}
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION, CATALYTIC ACTIVITY,
RP BIOPHYSICOCHEMICAL PROPERTIES, SUBUNIT, AND BIOTECHNOLOGY.
RC STRAIN=AHK119 {ECO:0000303|PubMed:20393707};
RX PubMed=20393707; DOI=10.1007/s00253-010-2555-x;
RA Hu X., Thumarat U., Zhang X., Tang M., Kawai F.;
RT "Diversity of polyester-degrading bacteria in compost and molecular
RT analysis of a thermoactive esterase from Thermobifida alba AHK119.";
RL Appl. Microbiol. Biotechnol. 87:771-779(2010).
RN [2] {ECO:0000305}
RP FUNCTION, CATALYTIC ACTIVITY, COFACTOR, ACTIVITY REGULATION,
RP BIOPHYSICOCHEMICAL PROPERTIES, BIOTECHNOLOGY, AND MUTAGENESIS OF ARG-67;
RP ALA-68; SER-69; ASN-126 AND ARG-137.
RX PubMed=22183084; DOI=10.1007/s00253-011-3781-6;
RA Thumarat U., Nakamura R., Kawabata T., Suzuki H., Kawai F.;
RT "Biochemical and genetic analysis of a cutinase-type polyesterase from a
RT thermophilic Thermobifida alba AHK119.";
RL Appl. Microbiol. Biotechnol. 95:419-430(2012).
RN [3] {ECO:0000305}
RP FUNCTION, CATALYTIC ACTIVITY, AND INDUCTION.
RX PubMed=25910960; DOI=10.1016/j.jbiosc.2015.03.006;
RA Thumarat U., Kawabata T., Nakajima M., Nakajima H., Sugiyama A., Yazaki K.,
RA Tada T., Waku T., Tanaka N., Kawai F.;
RT "Comparison of genetic structures and biochemical properties of tandem
RT cutinase-type polyesterases from Thermobifida alba AHK119.";
RL J. Biosci. Bioeng. 120:491-497(2015).
RN [4] {ECO:0000305}
RP FUNCTION, CATALYTIC ACTIVITY, BIOTECHNOLOGY, AND MUTAGENESIS OF TYR-99;
RP SER-169; TRP-194; ASP-215; ILE-217 AND HIS-247.
RX PubMed=33387709; DOI=10.1016/j.biortech.2020.124609;
RA Zhang Z., Wang W., Li D., Xiao J., Wu L., Geng X., Wu G., Zeng Z., Hu J.;
RT "Decolorization of molasses alcohol wastewater by thermophilic hydrolase
RT with practical application value.";
RL Bioresour. Technol. 323:124609-124609(2021).
RN [5] {ECO:0007744|PDB:3VIS}
RP X-RAY CRYSTALLOGRAPHY (1.76 ANGSTROMS) IN COMPLEX WITH CALCIUM, COFACTOR,
RP SUBUNIT, AND DISULFIDE BONDS.
RX DOI=10.1016/j.polymdegradstab.2012.02.003;
RA Kitadokoro K., Thumarat U., Nakamura R., Nishimura K., Karatani H.,
RA Suzuki H., Kawai F.;
RT "Crystal structure of cutinase Est119 from Thermobida alba AHK119 that can
RT degrade modpolyethylene terephthalate at 1.76 A resolution.";
RL Polym. Degrad. Stab. 97:771-775(2012).
RN [6] {ECO:0007744|PDB:3WYN}
RP X-RAY CRYSTALLOGRAPHY (1.68 ANGSTROMS) IN COMPLEX WITH CALCIUM, AND
RP DISULFIDE BONDS.
RA Kitadokoro K., Thumarat U., Kawai F.;
RT "Structure of calcium bound cutinase Est119 from Thermobifida alba.";
RL Submitted (SEP-2014) to the PDB data bank.
RN [7] {ECO:0007744|PDB:6AID}
RP X-RAY CRYSTALLOGRAPHY (1.30 ANGSTROMS) OF 35-300 OF MUTANT A-169 IN COMPLEX
RP WITH LACTIC ACID AND CALCIUM, COFACTOR, ACTIVE SITE, DISULFIDE BONDS, AND
RP REACTION MECHANISM.
RX PubMed=30761732; DOI=10.1111/febs.14781;
RA Kitadokoro K., Kakara M., Matsui S., Osokoshi R., Thumarat U., Kawai F.,
RA Kamitani S.;
RT "Structural insights into the unique polylactate-degrading mechanism of
RT Thermobifidaalba cutinase.";
RL FEBS J. 286:2087-2098(2019).
CC -!- FUNCTION: Catalyzes the hydrolysis of cutin, a polyester that forms the
CC structure of plant cuticle (PubMed:20393707, PubMed:22183084,
CC PubMed:25910960, PubMed:33387709). Shows esterase activity towards p-
CC nitrophenol-linked aliphatic esters (pNP-aliphatic esters)
CC (PubMed:20393707, PubMed:22183084, PubMed:25910960, PubMed:33387709).
CC Capable of degrading the plastic poly(ethylene terephthalate) (PET),
CC the most abundant polyester plastic in the world (By similarity). Can
CC also depolymerize the synthetic polyesters poly(epsilon-caprolactone)
CC (PCL), poly(butylene succinate-co-adipate) (PBSA), poly(butylene
CC succinate) (PBS), and poly(lactic acid) (PLA) (PubMed:20393707,
CC PubMed:22183084). {ECO:0000250|UniProtKB:D4Q9N1,
CC ECO:0000269|PubMed:20393707, ECO:0000269|PubMed:22183084,
CC ECO:0000269|PubMed:25910960, ECO:0000269|PubMed:33387709}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=an acetyl ester + H2O = acetate + an aliphatic alcohol + H(+);
CC Xref=Rhea:RHEA:12957, ChEBI:CHEBI:2571, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:30089, ChEBI:CHEBI:47622;
CC Evidence={ECO:0000269|PubMed:20393707, ECO:0000269|PubMed:22183084};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:12958;
CC Evidence={ECO:0000269|PubMed:20393707, ECO:0000269|PubMed:22183084};
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:0000305|PubMed:20393707, ECO:0000305|PubMed:22183084};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:49529;
CC Evidence={ECO:0000305|PubMed:20393707, ECO:0000305|PubMed:22183084};
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:20393707, ECO:0000269|PubMed:22183084,
CC ECO:0000269|PubMed:25910960, ECO:0000269|PubMed:33387709};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:47349;
CC Evidence={ECO:0000269|PubMed:20393707, ECO:0000269|PubMed:22183084,
CC ECO:0000269|PubMed:25910960, ECO:0000269|PubMed:33387709};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=cutin + H2O = cutin monomers.; EC=3.1.1.74;
CC Evidence={ECO:0000305|PubMed:20393707, ECO:0000305|PubMed:22183084,
CC ECO:0000305|PubMed:25910960, ECO:0000305|PubMed:33387709};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=a hexanoate ester + H2O = an aliphatic alcohol + H(+) +
CC hexanoate; Xref=Rhea:RHEA:47352, ChEBI:CHEBI:2571, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:17120, ChEBI:CHEBI:87656;
CC Evidence={ECO:0000269|PubMed:20393707, ECO:0000269|PubMed:22183084};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:47353;
CC Evidence={ECO:0000269|PubMed:20393707, ECO:0000269|PubMed:22183084};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=an octanoate ester + H2O = an aliphatic alcohol + H(+) +
CC octanoate; Xref=Rhea:RHEA:47356, ChEBI:CHEBI:2571, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:25646, ChEBI:CHEBI:87657;
CC Evidence={ECO:0000269|PubMed:20393707, ECO:0000269|PubMed:22183084};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:47357;
CC Evidence={ECO:0000269|PubMed:20393707, ECO:0000269|PubMed:22183084};
CC -!- COFACTOR:
CC Name=Ca(2+); Xref=ChEBI:CHEBI:29108;
CC Evidence={ECO:0000269|PubMed:30761732, ECO:0000269|Ref.5,
CC ECO:0000305|PubMed:22183084};
CC Note=Can also bind other divalent metal ions with lower efficiency
CC (PubMed:22183084). Calcium ion binding contributes to the
CC thermostability of the protein (PubMed:22183084).
CC {ECO:0000269|PubMed:22183084};
CC -!- ACTIVITY REGULATION: Activated by calcium ions (PubMed:22183084).
CC Activated by magnesium ions (PubMed:22183084). Activated by manganese
CC ions (PubMed:22183084). Inhibited by the serine hydrolase inhibitor
CC phenylmethanesulfonyl fluoride (PMSF) (PubMed:22183084). Inhibited by
CC the chelator ethylenediaminetetraacetic acid (EDTA) (PubMed:22183084).
CC Inhibited by iron ions (PubMed:22183084). Inhibited by aluminum ions
CC (PubMed:22183084). Inhibited by rubidium ions (PubMed:22183084).
CC Inhibited by lithium ions (PubMed:22183084).
CC {ECO:0000269|PubMed:22183084}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=3.22 mM for pNP-acetate (at 37 degrees Celsius and pH 7)
CC {ECO:0000269|PubMed:22183084};
CC KM=3.41 mM for pNP-butyrate (at 37 degrees Celsius and pH 7)
CC {ECO:0000269|PubMed:22183084};
CC KM=2.37 mM for pNP-hexanoate (at 37 degrees Celsius and pH 7)
CC {ECO:0000269|PubMed:22183084};
CC KM=1.87 mM for pNP-octanoate (at 37 degrees Celsius and pH 7)
CC {ECO:0000269|PubMed:22183084};
CC Note=kcat is 2.06 sec(-1) with pNP-acetate as substrate (at 37
CC degrees Celsius and pH 7) (PubMed:22183084). kcat is 4.48 sec(-1)
CC with pNP-butyrate as substrate (at 37 degrees Celsius and pH 7)
CC (PubMed:22183084). kcat is 3.28 sec(-1) with pNP-hexanoate as
CC substrate (at 37 degrees Celsius and pH 7) (PubMed:22183084). kcat is
CC 2.45 sec(-1) with pNP-octanoate as substrate (at 37 degrees Celsius
CC and pH 7) (PubMed:22183084). {ECO:0000269|PubMed:22183084};
CC pH dependence:
CC Optimum pH is 6. {ECO:0000269|PubMed:20393707};
CC Temperature dependence:
CC Optimum temperature is 50 degrees Celsius.
CC {ECO:0000269|PubMed:20393707};
CC -!- SUBUNIT: Monomer. {ECO:0000269|PubMed:20393707, ECO:0000269|Ref.5}.
CC -!- SUBCELLULAR LOCATION: Secreted {ECO:0000250|UniProtKB:G8GER6}.
CC Periplasm {ECO:0000250|UniProtKB:G8GER6}.
CC -!- INDUCTION: Expression is not induced by the synthetic polyester
CC poly(butylene succinate-co-adipate) (PBSA).
CC {ECO:0000269|PubMed:25910960}.
CC -!- BIOTECHNOLOGY: Shows promising applications in ethyl alcohol distillery
CC processes as it may be utilized for treatment of molasses wastewater
CC (PubMed:33387709). Has potential for application in biological
CC recycling of plastic waste products (PubMed:20393707, PubMed:22183084).
CC {ECO:0000269|PubMed:20393707, ECO:0000269|PubMed:22183084,
CC ECO:0000269|PubMed:33387709}.
CC -!- SIMILARITY: Belongs to the AB hydrolase superfamily. {ECO:0000305}.
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DR EMBL; AB445476; BAK48590.1; -; Genomic_DNA.
DR PDB; 3VIS; X-ray; 1.76 A; A/B=1-300.
DR PDB; 3WYN; X-ray; 1.68 A; A/B=1-300.
DR PDB; 6AID; X-ray; 1.30 A; A=35-300.
DR PDBsum; 3VIS; -.
DR PDBsum; 3WYN; -.
DR PDBsum; 6AID; -.
DR SMR; F7IX06; -.
DR ESTHER; 9acto-f7ix06; Polyesterase-lipase-cutinase.
DR SABIO-RK; F7IX06; -.
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:0046872; F:metal ion binding; IEA:UniProtKB-KW.
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 3D-structure; Calcium; Disulfide bond; Hydrolase; Metal-binding; Periplasm;
KW Secreted; Serine esterase; Signal.
FT SIGNAL 1..39
FT /evidence="ECO:0000255"
FT CHAIN 40..300
FT /note="Cutinase est2"
FT /evidence="ECO:0000255"
FT /id="PRO_5003363093"
FT ACT_SITE 169
FT /note="Nucleophile"
FT /evidence="ECO:0000305|PubMed:30761732"
FT ACT_SITE 215
FT /note="Charge relay system"
FT /evidence="ECO:0000305|PubMed:30761732"
FT ACT_SITE 247
FT /note="Charge relay system"
FT /evidence="ECO:0000305|PubMed:30761732"
FT BINDING 99
FT /ligand="poly(ethylene terephthalate)"
FT /ligand_id="ChEBI:CHEBI:131701"
FT /evidence="ECO:0000250|UniProtKB:A0A0K8P6T7"
FT BINDING 170
FT /ligand="poly(ethylene terephthalate)"
FT /ligand_id="ChEBI:CHEBI:131701"
FT /evidence="ECO:0000250|UniProtKB:A0A0K8P6T7"
FT BINDING 194
FT /ligand="poly(ethylene terephthalate)"
FT /ligand_id="ChEBI:CHEBI:131701"
FT /evidence="ECO:0000250|UniProtKB:A0A0K8P6T7"
FT BINDING 213
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /evidence="ECO:0000269|PubMed:30761732,
FT ECO:0007744|PDB:3WYN, ECO:0007744|PDB:6AID"
FT BINDING 243
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /evidence="ECO:0000269|PubMed:30761732,
FT ECO:0007744|PDB:3WYN, ECO:0007744|PDB:6AID"
FT BINDING 292
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /evidence="ECO:0000269|PubMed:30761732,
FT ECO:0007744|PDB:3WYN, ECO:0007744|PDB:6AID"
FT DISULFID 280..298
FT /evidence="ECO:0007744|PDB:3VIS, ECO:0007744|PDB:3WYN"
FT MUTAGEN 67
FT /note="R->C: Increases activity on pNP-butyrate."
FT /evidence="ECO:0000269|PubMed:22183084"
FT MUTAGEN 68
FT /note="A->V: Increases activity on pNP-butyrate."
FT /evidence="ECO:0000269|PubMed:22183084"
FT MUTAGEN 69
FT /note="S->P: Increases activity on pNP-butyrate; when
FT associated with C-137."
FT /evidence="ECO:0000269|PubMed:22183084"
FT MUTAGEN 99
FT /note="Y->A: Decreases activity; when associated with A-194
FT and A-217."
FT /evidence="ECO:0000269|PubMed:33387709"
FT MUTAGEN 126
FT /note="N->I: Increases activity on pNP-butyrate."
FT /evidence="ECO:0000269|PubMed:22183084"
FT MUTAGEN 137
FT /note="R->C: Increases activity on pNP-butyrate; when
FT associated with P-69."
FT /evidence="ECO:0000269|PubMed:22183084"
FT MUTAGEN 169
FT /note="S->A: Abolishes activity."
FT /evidence="ECO:0000269|PubMed:33387709"
FT MUTAGEN 194
FT /note="W->A: Decreases activity; when associated with A-99
FT and A-217."
FT /evidence="ECO:0000269|PubMed:33387709"
FT MUTAGEN 215
FT /note="D->A: Abolishes activity."
FT /evidence="ECO:0000269|PubMed:33387709"
FT MUTAGEN 217
FT /note="I->A: Decreases activity; when associated with A-99
FT and A-194."
FT /evidence="ECO:0000269|PubMed:33387709"
FT MUTAGEN 247
FT /note="H->A: Abolishes activity."
FT /evidence="ECO:0000269|PubMed:33387709"
SQ SEQUENCE 300 AA; 32353 MW; D7FC433E0E8FF936 CRC64;
MSVTTPRRET SLLSRALRAT AAAATAVVAT VALAAPAQAA NPYERGPNPT ESMLEARSGP
FSVSEERASR FGADGFGGGT IYYPRENNTY GAIAISPGYT GTQSSIAWLG ERIASHGFVV
IAIDTNTTLD QPDSRARQLN AALDYMLTDA SSAVRNRIDA SRLAVMGHSM GGGGTLRLAS
QRPDLKAAIP LTPWHLNKSW RDITVPTLII GAEYDTIASV TLHSKPFYNS IPSPTDKAYL
ELDGASHFAP NITNKTIGMY SVAWLKRFVD EDTRYTQFLC PGPRTGLLSD VEEYRSTCPF
