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Selective Hydrolysis by Engineered Cutinases: Characterization of Aliphatic-Aromatic Homo and Co-Polyesters by LC and
Eman Abdelraheem1, Vasilis Tseliou1, Jessica Desport1
1van 't Hoff Institute for Molecular Science, University of Amsterdam, Science Park 904, 1098, XH Amsterdam, the Netherlands.
Chemistry (Weinheim an Der Bergstrasse, Germany)
|February 12, 2025
Summary
Engineered enzymes efficiently break down synthetic polyesters, enabling polymer recycling. This research highlights enzyme applications in analyzing and degrading various polymer types under mild conditions.
Area of Science:
- Biotechnology
- Polymer Science
- Enzymology
Background:
- Polymer properties like biodegradability and recyclability are tunable via monomer selection.
- Hydrolases offer a mild alternative to chemical methods for polyester depolymerization.
Purpose of the Study:
- To engineer a thermostable cutinase (Tc_Cut2NVWCCG) from Thermobifida cellulosilytica.
- To compare the hydrolysis efficiency of engineered Tc_Cut2NVWCCG and leaf-branch compost cutinase (LCCWCCG) on various polyesters.
- To investigate the enzymatic degradation mechanism of aliphatic/aromatic co-polyesters.
Main Methods:
- Enzyme engineering for enhanced thermostability.
- Enzymatic hydrolysis assays on low molar mass substrates, homo-polyesters, and co-polyesters.
- Size exclusion chromatography coupled with mass spectrometry (SEC-MS) for analyzing hydrolysis products and molecular weight changes.
Main Results:
- Engineered Tc_Cut2NVWCCG exhibited improved thermostability up to 91°C.
- Both enzymes showed higher hydrolysis rates for aliphatic than aromatic homo-polyesters.
- Enzymatic hydrolysis of co-polyesters occurred selectively at aliphatic monomers, reducing molecular weight and altering end-group composition.
Conclusions:
- Enzyme engineering can yield highly active enzymes for synthetic polyester hydrolysis.
- Co-polymer composition significantly influences biodegradation rates.
- Enzymes can be utilized for the analytical characterization of synthetic polymers through selective molecular weight reduction.
- Developed enzymes offer potential for efficient and mild polyester recycling.
