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Multi-enzyme Screening Using a High-throughput Genetic Enzyme Screening System

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Surface display as a functional screening platform for detecting enzymes active on PET.

Sophia A H Heyde^1,2, Jenny Arnling Bååth³, Peter Westh³

¹Novozymes A/S, Biologiens Vej 2, 2800, Kgs. Lyngby, Denmark.

Microbial Cell Factories

|May 2, 2021

Summary

Researchers engineered a surface display system for PETase enzymes in bacteria. This innovation enables faster screening of PET-degrading enzymes, advancing plastic biodegradation research.

Keywords:

E. coli Extracellular protein production Ideonella sakaiensis PETase Surface display

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Area of Science:

Biotechnology
Environmental Science
Polymer Science

Background:

Poly(ethylene terephthalate) (PET) is a prevalent plastic pollutant, with current recycling methods degrading its quality.
Existing PET biodegradation methods using PETase enzymes are slow and hindered by inefficient screening processes.
Developing sustainable plastic degradation solutions is crucial for environmental remediation.

Purpose of the Study:

To develop a more efficient bacterial screening platform for engineering improved PETase enzymes.
To investigate the efficacy of different surface display anchors for functional PETase expression in E. coli.
To facilitate the high-throughput screening of PETase variants for enhanced plastic hydrolysis.

Main Methods:

Functional display of PETase enzymes on the surface of Escherichia coli using various anchor proteins.
Assaying PETase activity by monitoring the hydrolysis of PET microparticles.
Comparing enzyme activity of surface-displayed PETase with solubilized enzyme preparations.

Main Results:

Successful functional display of active PETase enzymes on the bacterial cell surface was achieved.
The surface display system enabled direct screening of enzyme activity on PET microparticles.
This platform allows for the assessment of both cell-anchored and solubilized PETase activity.

Conclusions:

Bacterial surface display provides an efficient platform for screening and engineering PETase enzymes.
This method accelerates the development of more effective biocatalysts for PET biodegradation.
The developed system supports the advancement of sustainable plastic recycling technologies.