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

• Published in: Angewandte Chemie International Edition 2024-11, Vol.63 (47), p.e202408979-n/a
• Main Authors: Yarman, Aysu, Waffo, Armel F. T., Katz, Sagie, Bernitzky, Cornelius, Kovács, Norbert, Borrero, Paloma, Frielingsdorf, Stefan, Supala, Eszter, Dragelj, Jovan, Kurbanoglu, Sevinc, Neumann, Bettina, Lenz, Oliver, Mroginski, Maria Andrea, Gyurcsányi, Róbert E., Wollenberger, Ulla, Scheller, Frieder W., Caserta, Giorgio, Zebger, Ingo
• Format: Article
• Language: English
• Subjects: Alkaline phosphatase; Alkaline Phosphatase - chemistry; Alkaline Phosphatase - metabolism; Binding sites; Biocatalysis; Biocompatibility; Biotechnology; Catalysis; Electrochemical Techniques; Electrochemistry; Enzymes; Epitopes; Exploitation; Hydrogenase; Hydrogenase - chemistry; Hydrogenase - metabolism; Imprinted polymers; Infrared spectroscopy; Iron compounds; Molecular dynamics; Molecular Dynamics Simulation; Molecular Dynamics Simulations; Molecularly Imprinted Polymers; Molecularly Imprinted Polymers - chemistry; Nickel compounds; Polymers; Polymers - chemistry; Proteins; Surface Enhanced Infrared Absorption Spectroscopy
• ISSN: 1433-7851; 1521-3773; 1521-3773
• DOI: 10.1002/anie.202408979

Molecularly imprinted polymers (MIPs) are artificial receptors equipped with selective recognition sites for target molecules. One of the most promising strategies for protein MIPs relies on the exploitation of short surface‐exposed protein fragments, termed epitopes, as templates to imprint binding sites in a polymer scaffold for a desired protein. However, the lack of high‐resolution structural data of flexible surface‐exposed regions challenges the selection of suitable epitopes. Here, we addressed this drawback by developing a polyscopoletin‐based MIP that recognizes recombinant proteins via imprinting of the widely used Strep‐tag II affinity peptide (Strep‐MIP). Electrochemistry, surface‐sensitive IR spectroscopy, and molecular dynamics simulations were employed to ensure an utmost control of the Strep‐MIP electrosynthesis. The functionality of this novel platform was verified with two Strep‐tagged enzymes: an O2‐tolerant [NiFe]‐hydrogenase, and an alkaline phosphatase. The enzymes preserved their biocatalytic activities after multiple utilization confirming the efficiency of Strep‐MIP as a general biocompatible platform to confine recombinant proteins for exploitation in biotechnology. The universal platform Strep‐MIP, built using the synthetic polymer‐based imprinting strategy, recognizes recombinant proteins via the widely used Strep‐tag II affinity peptide. Electrochemistry, surface‐sensitive IR spectroscopy, and molecular dynamics simulations provide comprehensive details of the binding, removal, reusability, and the catalytic performance of the immobilized enzymes, fostering the application of Strep‐MIP in biotechnology.