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A Strep-Tag Imprinted Polymer Platform for Heterogenous Bio(electro)catalysis.

Aysu Yarman^1,2, Armel F T Waffo³, Sagie Katz³

¹Institute of Biochemistry and Biology, University of Potsdam, Karl-Liebknecht Str. 24-25, 14476, Potsdam.

Angewandte Chemie (International Ed. in English)

|July 9, 2024

Summary

This study introduces a novel molecularly imprinted polymer (MIP) for recognizing proteins using the Strep-tag II affinity peptide. This biocompatible platform efficiently immobilizes enzymes, preserving their activity for biotechnological applications.

Keywords:

Biocatalysis Electrochemistry Molecular Dynamics Simulations Molecularly Imprinted Polymers Surface Enhanced Infrared Absorption Spectroscopy

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Area of Science:

Biomaterials Science
Protein Engineering
Analytical Chemistry

Background:

Molecularly imprinted polymers (MIPs) are synthetic receptors with tailored recognition sites.
Protein MIPs often utilize surface-exposed epitopes as templates, but structural data limitations hinder epitope selection.
Developing robust MIPs for specific protein recognition remains a challenge.

Purpose of the Study:

To develop a polyscopoletin-based MIP for recognizing recombinant proteins via the Strep-tag II affinity peptide (Strep-MIP).
To overcome challenges in selecting suitable epitopes for protein MIPs by using a common affinity tag.
To create a general and biocompatible platform for immobilizing and utilizing recombinant proteins.

Main Methods:

Electrosynthesis of Strep-MIP using polyscopoletin.
Characterization using electrochemistry and surface-sensitive IR spectroscopy.
Validation through molecular dynamics simulations and binding assays with Strep-tagged enzymes.

Main Results:

Successful development of a Strep-tag II-imprinted MIP (Strep-MIP).
Demonstrated recognition and immobilization of two Strep-tagged enzymes: O2-tolerant [NiFe]-hydrogenase and alkaline phosphatase.
Enzymes retained biocatalytic activity after multiple uses, confirming the platform's efficiency and biocompatibility.

Conclusions:

Strep-MIP provides a reliable and biocompatible method for imprinting and confining Strep-tagged recombinant proteins.
This platform facilitates the exploitation of enzymes in various biotechnological applications.
The approach offers a solution for challenges associated with epitope selection in protein MIP design.