Affiliation:
1. Functional and Interactive Polymers Institute of Technical and Macromolecular Chemistry RWTH Aachen University Worringerweg 2 52074 Aachen Germany
2. DWI − Leibniz‐Institute for Interactive Materials e.V. Forckenbeckstraße 50 52074 Aachen Germany
3. Laboratory for Biomaterials Institute of Biotechnology and Helmholtz‐Institute for Biomedical Engineering RWTH Aachen University Pauwelsstraße 20 52074 Aachen Germany
4. Aachen Maastricht Institute for Biobased Materials (AMIBM) Maastricht University Brightlands Chemelot Campus Urmonderbaan 22 RD Geleen 6167 The Netherlands
Abstract
AbstractEnzymes play a vital role in synthesizing complex biological molecules like hyaluronic acid (HA). Immobilizing enzymes on support materials is essential for their efficient use and reuse in multiple cycles. Microgels, composed of cross–linked, highly swollen polymer networks, are ideal for enzyme uptake owing to their high porosity. This study demonstrates the immobilization of His6‐tagged hyaluronan synthase from Pasteurella multocida (PmHAS) onto nitrilotriacetic acid functionalized microgels using different bivalent ions (Ni2+, Co2+, Mn2+, Mg2+, and Fe2+) via metal affinity binding. The results indicate that using Ni2+ yields the microgels with the highest enzyme uptake and HA formation. The immobilized PmHAS enables repetitive enzymatic production, producing high molecular weight HAs with decreasing dispersities in each step. Furthermore, the highest reported yield of HA with high molecular weight for immobilized PmHAS is achieved. This system establishes a foundation for continuous HA formation, with future works potentially enhancing PmHAS stability through protein engineering.
Funder
European Commission
Bundesministerium für Bildung und Forschung