A versatile assay platform for enzymatic poly(ethylene-terephthalate) degradation

Area of Science:

• Biotechnology
• Environmental Science
• Biochemistry

Background:

• Plastic accumulation, particularly microplastics, poses a significant global environmental threat.
• The discovery of polyethylene terephthalate (PET)-degrading enzymes offers novel strategies for plastic decomposition and recycling.
• Existing methods for identifying and characterizing PET-degrading enzymes lack the speed and reliability needed for extensive natural diversity exploration and protein engineering.

Purpose of the Study:

• To develop a fast, reliable, and versatile assay platform for the discovery and characterization of PET-degrading enzymes.
• To facilitate protein engineering and directed evolution studies for enhanced enzyme performance.
• To enable both high-precision analysis and high-throughput screening of enzyme activity.

Main Methods:

• Coating standard laboratory consumables (PCR tubes, microtiter plates) with PET to create a substrate with adjustable crystallinity.
• Integrating the PET-coated consumables with ultra-high performance liquid chromatography (UHPLC) or fluorometric detection systems.
• Implementing enzyme quantification methods alongside activity assays.

Main Results:

• A versatile assay platform was successfully established using PET-coated lab consumables.
• The platform demonstrated adaptability for both detailed, high-precision analyses and high-throughput screening applications.
• The system is suitable for analyzing crude enzyme lysates, broadening its applicability.

Conclusions:

• The developed assay platform provides a crucial tool for exploring the diversity of PET-degrading enzymes.
• This innovation supports advancements in biological plastic decomposition and technical recycling solutions.
• The platform's versatility enhances the efficiency of enzyme discovery and engineering processes.