title: Cleavage Product Accumulation Decreases the Activity of Cutinase during PET Hydrolysis.
abstract: The Fusarium solani cutinase (FsC) is a promising candidate for the enzymatic degradation of the synthetic polyester polyethylene terephthalate (PET) but still suffers from a lack of activity. Using atomic MD simulations with different concentrations of cleavage product ethylene glycol (EG), we show influences of EG on the dynamic of FsC. We observed accumulation of EG in the active site region reducing the local flexibility of FsC. Furthermore, we used a coarse-grained mechanical model to investigate whether substrate binding in the active site causes an induced fit. We observed this supposed induced fit or "breath-like" movement during substrate binding indicating that the active site has to be flexible for substrate conversion. This guides rational design: mutants with an increased flexibility near the active site should be considered to compensate the solvent-mediated reduction in activity.
authors.affiliation: Department of Biology, Computational Biology & Simulation Group, Technische Universität Darmstadt , Schnittspahnstraße 2, 64287 Darmstadt, Germany.
authors.forename: Christine
authors.identifier: 0000-0002-4600-9356
authors.initials: C
authors.lastname: Groß
authors.affiliation: Department of Biology, Computational Biology & Simulation Group, Technische Universität Darmstadt , Schnittspahnstraße 2, 64287 Darmstadt, Germany.
authors.forename: Kay
authors.initials: K
authors.lastname: Hamacher
authors.affiliation: Department of Chemistry, Biological Chemistry Group, Technische Universität Darmstadt , Alarich-Weiss-Straße 8, 64287 Darmstadt, Germany.
authors.forename: Katja
authors.initials: K
authors.lastname: Schmitz
authors.affiliation: Department of Biology, Computational Biology & Simulation Group, Technische Universität Darmstadt , Schnittspahnstraße 2, 64287 Darmstadt, Germany.
authors.forename: Sven
authors.initials: S
authors.lastname: Jager
mesh_terms: D055162:Biocatalysis; D001673:Biodegradation, Environmental; D002265:Carboxylic Ester Hydrolases / Q000737:chemistry / Q000378:metabolism*; D020134:Catalytic Domain; D005670:Fusarium / Q000201:enzymology / Q000378:metabolism; D006868:Hydrolysis; D056004:Molecular Dynamics Simulation; D011093:Polyethylene Terephthalates / Q000302:isolation & purification / Q000378:metabolism*
chemical_list: D011093:Polyethylene Terephthalates; D002265:Carboxylic Ester Hydrolases; C019952:cutinase