Information on EC 3.1.1.101 - poly(ethylene terephthalate) hydrolase

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The expected taxonomic range for this enzyme is: Eukaryota, Bacteria

EC NUMBER
COMMENTARY hide
3.1.1.101
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RECOMMENDED NAME
GeneOntology No.
poly(ethylene terephthalate) hydrolase
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
(ethylene terephthalate)n + H2O = (ethylene terephthalate)n-1 + 4-[(2-hydroxyethoxy)carbonyl]benzoate
show the reaction diagram
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
polyethylene terephthalate degradation
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SYSTEMATIC NAME
IUBMB Comments
poly(ethylene terephthalate) hydrolase
The enzyme, isolated from the bacterium Ideonella sakaiensis, also produces small amounts of terephthalate (cf. EC 3.1.1.102, mono(ethylene terephthalate) hydrolase). The reaction takes place on PET-film placed in solution.
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(epsilon-caprolactone)n + H2O
(epsilon-caprolactame)n-1 + ethylene terephthalate
show the reaction diagram
(ethylene terephthalate)n + H2O
(ethylene terephthalate)n-1 + ethylene terephthalate
show the reaction diagram
(ethylene terephthalate)n + H2O
(ethylene terephthalate)n-1 + mono(2-hydroxyethyl)terephthalic acid
show the reaction diagram
4-nitrophenyl butanoate + H2O
4-nitrophenol + butanoate
show the reaction diagram
bis(2-hydroxyethyl) terephthalic acid + H2O
mono(2-hydroxyethyl)terephthalic acid + ethylene glycol
show the reaction diagram
additional information
?
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ca2+
and Mg2+, presence of 10 mM each increases thermostability by about 10 degrees. Residues Asp174, Asp204, and Glu253 are potential binding residues for the two cations; and Mg2+, presence of 10 mM each increases thermostability by about 14 degrees
Mg2+
and Ca2+, presence of 10 mM each increases thermostability by about 10 degrees; and Ca2+, presence of 10 mM each increases thermostability by about 10 degrees. Residues Asp174, Asp204, and Glu253 are potential binding residues for the two cations
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
bis(2-hydroxyethyl) terephthalic acid
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competitive inhibition of the hydrolytic activity against PET nanoparticles
ethylene terephthalate
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competitive inhibition of the hydrolytic activity against PET nanoparticles
mono-(2-hydroxyethyl) terephthalate
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strong inhibition
PMSF
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the PMSF protein complex reveals covalent modification of S130 and binding of one of the oxygens of the tetrahedral adduct in the oxyanion hole formed by the main chain nitrogens of M131 and Y60
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2.3 - 12.4
4-nitrophenyl butanoate
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
80 - 170
MOPS
240 - 440
Tris
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
9.5
60% of maximum activity; 60% of maximum activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
65 - 70
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wild-type
75 - 80
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mutant D204C/E253C/D174R
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
PDB
SCOP
CATH
ORGANISM
UNIPROT
Ideonella sakaiensis (strain 201-F6)
Ideonella sakaiensis (strain 201-F6)
Ideonella sakaiensis (strain 201-F6)
Ideonella sakaiensis (strain 201-F6)
Ideonella sakaiensis (strain 201-F6)
Ideonella sakaiensis (strain 201-F6)
Ideonella sakaiensis (strain 201-F6)
Ideonella sakaiensis (strain 201-F6)
Ideonella sakaiensis (strain 201-F6)
Ideonella sakaiensis (strain 201-F6)
Ideonella sakaiensis (strain 201-F6)
Ideonella sakaiensis (strain 201-F6)
Ideonella sakaiensis (strain 201-F6)
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
molecular replacement with a poly-Ala model of Streptomyces exofoliatus lipase, PDB-ID: 1JFR, in free as well as in inhibitor-bound form. The enzyme forms a classical alpha/beta-hydrolase fold with a central nine-stranded beta-sheet flanked by 11 alpha-helices on both sides. The catalytic triad, comprising S130, D176 and H208, is located in a crevice on the surface of the enzyme
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structural model generated on the basis of PDB ID 3VIS. The polyester hydrolase reveals a typical alpha/beta hydrolase fold. The catalytic triad formed by residues S130, D176 and H208 is exposed to the solvent; structural model generated on the basis of PDB ID 3VIS. The polyester hydrolase reveals a typical alpha/beta hydrolase fold. The catalytic triad formed by residues S130, D176 and H208 is exposed to the solvent
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
50
60 min, 40% residual activity
55
60 min, 80% residual activity
60
10 min, 15% residual activity; 10 min, 35% residual activity
61
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loss of activity above
69.8
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melting temperature, wild-type
70
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melting temperature
71.2
melting point; melting point
84.6
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melting temperature, wild-type, presence of 10 mM Ca2+
88.3
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melting temperature, mutant D174C/E253C; melting temperature, mutant D204C/E253C
91.6
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melting temperature, mutant D174C/E253C, presence of 10 mM Ca2+
additional information
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expression in Escherichia coli; expression in Escherichia coli
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D174C/D253C
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increase in melting temperature in absence and in presence of Ca2+
D204C/E253C
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increase in melting temperature
D204C/E253C/D174R
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increase in temperature optimum to 75-80°C, mutant causes a weight loss of PET films of 25.0% at 70 °C after a reaction time of 48 h, compared to 0.3% for wild-type
D204R
mutation in residue involved in metal ion binding. Increase in melting point by 14 degrees compared to wild-type. Presence of 10 mM CaCl2 does not result in a considerable increase in melting point
E253R
mutation in residue involved in metal ion binding. Increase in melting point by 14 degrees compared to wild-type. Presence of 10 mM CaCl2 does not result in a considerable increase in melting point
G62A
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exchange of amino acid residues of TfCut2 involved in substrate binding with those present in LC-cutinase, UniProt ID G9BY57, from an uncultured bacterium, leads to enzyme variants with increased PET hydrolytic activity at 65°C. Variant causes a weight loss of PET films of more than 42% after 50 h of hydrolysis, corresponding to a 2.7fold increase compared to the wild type enzyme. Mutant G62A reveals a 5.5fold lower binding constant to the inhibitor mono-(2-hydroxyethyl) terephthalate than the wild type enzyme
G62A/I213S
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exchange of amino acid residues of TfCut2 involved in substrate binding with those present in LC-cutinase, UniProt ID G9BY57, from an uncultured bacterium, leads to enzyme variants with increased PET hydrolytic activity at 65°C. Variant causes a weight loss of PET films of more than 42% after 50 h of hydrolysis, corresponding to a 2.7fold increase compared to the wild type enzyme
D174C/D253C
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increase in melting temperature in absence and in presence of Ca2+
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D204C/E253C
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increase in melting temperature
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D204C/E253C/D174R
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increase in temperature optimum to 75-80°C, mutant causes a weight loss of PET films of 25.0% at 70 °C after a reaction time of 48 h, compared to 0.3% for wild-type
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D204R
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mutation in residue involved in metal ion binding. Increase in melting point by 14 degrees compared to wild-type. Presence of 10 mM CaCl2 does not result in a considerable increase in melting point
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E253R
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mutation in residue involved in metal ion binding. Increase in melting point by 14 degrees compared to wild-type. Presence of 10 mM CaCl2 does not result in a considerable increase in melting point
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G62A
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exchange of amino acid residues of TfCut2 involved in substrate binding with those present in LC-cutinase, UniProt ID G9BY57, from an uncultured bacterium, leads to enzyme variants with increased PET hydrolytic activity at 65°C. Variant causes a weight loss of PET films of more than 42% after 50 h of hydrolysis, corresponding to a 2.7fold increase compared to the wild type enzyme. Mutant G62A reveals a 5.5fold lower binding constant to the inhibitor mono-(2-hydroxyethyl) terephthalate than the wild type enzyme
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G62A/I213S
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exchange of amino acid residues of TfCut2 involved in substrate binding with those present in LC-cutinase, UniProt ID G9BY57, from an uncultured bacterium, leads to enzyme variants with increased PET hydrolytic activity at 65°C. Variant causes a weight loss of PET films of more than 42% after 50 h of hydrolysis, corresponding to a 2.7fold increase compared to the wild type enzyme
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APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
degradation