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Literature summary extracted from

  • Wei, R.; Oeser, T.; Schmidt, J.; Meier, R.; Barth, M.; Then, J.; Zimmermann, W.
    Engineered bacterial polyester hydrolases efficiently degrade polyethylene terephthalate due to relieved product inhibition (2016), Biotechnol. Bioeng., 113, 1658-1665.
    View publication on PubMed

Application

EC Number Application Comment Organism
3.1.1.101 degradation 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 Thermobifida fusca

Protein Variants

EC Number Protein Variants Comment Organism
3.1.1.101 G62A 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 Thermobifida fusca
3.1.1.101 G62A/I213S 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 Thermobifida fusca

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.1.1.101 mono-(2-hydroxyethyl) terephthalate strong inhibition Thermobifida fusca

Organism

EC Number Organism UniProt Comment Textmining
3.1.1.101 Thermobifida fusca E5BBQ3 cf. EC 3.1.1.74
-
3.1.1.101 Thermobifida fusca KW3 E5BBQ3 cf. EC 3.1.1.74
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.1.1.101 (ethylene terephthalate)n + H2O substrates are amorphous PET film or PET fibers Thermobifida fusca (ethylene terephthalate)n-1 + 4-[(2-hydroxyethoxy)carbonyl]benzoate terephthalate is the dominant hydrolysis product detected after a reaction time of 50 h at 65°C. Monohydroxyethylene terephthalate which inhibits the hydrolysis of PET is almost completely converted into terephthalate and accounts for less than 9.1% of the total hydrolysis products ?
3.1.1.101 (ethylene terephthalate)n + H2O substrates are amorphous PET film or PET fibers Thermobifida fusca KW3 (ethylene terephthalate)n-1 + 4-[(2-hydroxyethoxy)carbonyl]benzoate terephthalate is the dominant hydrolysis product detected after a reaction time of 50 h at 65°C. Monohydroxyethylene terephthalate which inhibits the hydrolysis of PET is almost completely converted into terephthalate and accounts for less than 9.1% of the total hydrolysis products ?