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Design, Production, and Characterization of Catalytically Active Inclusion Bodies.

Gizem Ölçücü^1,2, Karl-Erich Jaeger^3,4, Ulrich Krauss^5,6

¹Institute of Molecular Enzyme Technology, Heinrich Heine University Düsseldorf, Forschungszentrum Jülich, Wilhelm-Johnen-Straße, Jülich, Germany. g.olcucu@fz-juelich.de.

Methods in Molecular Biology (Clifton, N.J.)

|January 19, 2023

Summary

Catalytically active inclusion bodies (CatIBs) offer a novel approach to enzyme immobilization for biocatalysis and medicine. This work details the methods for designing, producing, and characterizing these protein-based catalysts.

Keywords:

Aggregation-inducing tag Biocatalysis Catalytically active inclusion bodies – CatIBs Enzyme aggregates Enzyme immobilization Fractionation Fusion protein Heterologous gene expression Microscopy

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Area of Science:

Biotechnology
Protein Engineering
Biocatalysis

Background:

Enzyme immobilization is crucial for industrial biocatalysis.
Biologically produced protein immobilizates offer sustainable alternatives.
Catalytically active inclusion bodies (CatIBs) are an emerging class of such immobilizates.

Purpose of the Study:

To provide a comprehensive methodology for the development of CatIBs.
To enable the application of CatIBs in biocatalysis, synthetic chemistry, and biomedicine.

Main Methods:

Designing fusion proteins with aggregation-inducing tags.
Heterologous protein production to induce CatIB formation.
Characterization techniques for assessing CatIB structure and activity.

Main Results:

Demonstrated successful design and production of CatIBs.
Established protocols for CatIB characterization.
Highlighted the potential of CatIBs as versatile biocatalytic tools.

Conclusions:

CatIBs represent a powerful platform for enzyme immobilization.
The presented methodology facilitates the creation and application of CatIBs.
CatIB technology holds significant promise for various scientific and industrial fields.