Catalytically Active Inclusion Bodies─Benchmarking and Application in Flow Chemistry
Gizem Ölçücü1,2, Benedikt Baumer3, Kira Küsters2
1Institute of Molecular Enzyme Technology, Heinrich Heine University Düsseldorf, Forschungszentrum Jülich GmbH, Wilhelm Johnen Strasse, D-52425 Jülich, Germany.
ACS Synthetic Biology
|May 2, 2022
Summary
We benchmarked tags for in vivo enzyme immobilization using catalytically active inclusion bodies (CatIBs). Tag selection and fusion strategy significantly impact CatIB yield, activity, and stability for industrial applications.
Area of Science:
- Biotechnology
- Protein Engineering
- Industrial Enzymology
Background:
- Enzyme immobilization is crucial for stable preparations in industrial batch and flow processes.
- Simultaneous enzyme production and in vivo immobilization strategies are emerging as alternatives to traditional methods.
- Catalytically active inclusion bodies (CatIBs) represent a promising in vivo enzyme immobilizate.
Purpose of the Study:
- To benchmark different aggregation-inducing tags and fusion strategies for CatIB formation.
- To compare the impact of tag selection on CatIB properties like yield, activity, and stability.
- To evaluate the application of optimized CatIBs in flow chemistry.
Main Methods:
- Fusion of various aggregation-inducing tags to target enzymes.
- Induction of CatIB formation in vivo.
- Characterization of CatIB properties (yield, activity, stability).
- Application of optimized CatIBs in a flow synthesis setup.
Main Results:
- Tag selection and fusion strategy significantly influence CatIB yield, activity, and stability.
- Optimization of CatIBs for alcohol dehydrogenase demonstrated superior activity and stability.
- First-time application of CatIBs in a flow synthesis approach was successful.
Conclusions:
- CatIB properties are highly dependent on the chosen tag and fusion strategy.
- Optimized CatIBs show significant potential for industrial enzyme applications.
- CatIBs are robust and effective for use in flow chemistry systems.
