Immobilization of enzymes provides many benefits, including facile separation and recovery of enzymes from reaction mixtures, enhanced stability, and co‐localization of multiple enzymes. Calcium‐phosphate‐protein supraparticles imbued with a leucine zipper binding domain (ZR) serve as a modular immobilization platform for enzymes fused to the complementary leucine zipper domain (ZE). The zippers provide high‐affinity, specific binding, separating enzymatic activity from the binding event. Using fluorescent model proteins (mCherryZEand eGFPZE), an amine dehydrogenase (AmDHZE), and a formate dehydrogenase (FDHZE), the efficacy of supraparticles as a biocatalytic solid support was assessed. Supraparticles demonstrated several benefits as an immobilization support, including predictable loading of multiple proteins, structural integrity in a panel of solvents, and the ability to elute and reload proteins without damaging the support. The dual‐enzyme reaction successfully converted ketone to amine on supraparticles, highlighting the efficacy of this system.
Enzyme immobilization is an essential technology for commercializing biocatalysis. It imparts stability, recoverability, and other valuable features that improve the effectiveness of biocatalysts. While many avenues to join an enzyme to solid phases exist, protein‐mediated immobilization is rapidly developing and has many advantages. Protein‐mediated immobilization allows for the binding interaction to be genetically coded, can be used to create artificial multienzyme cascades, and enables modular designs that expand the variety of enzymes immobilized. By designing around binding interactions between protein domains, they can be integrated into functional materials for protein immobilization. These materials are framed within the context of biocatalytic performance, immobilization efficiency, and stability of the materials. In this review, supports composed entirely of protein are discussed first, with systems such as cellulosomes and protein cages being discussed alongside newer technologies like spore‐based biocatalysts and forizymes. Protein‐composite materials such as polymersomes and protein–inorganic supraparticles are then discussed to demonstrate how protein‐mediated strategies are applied to many classes of solid materials. Critical analysis and future directions of protein‐based immobilization are then discussed, with a particular focus on both computational and design strategies to advance this area of research and make it more broadly applicable to many classes of enzymes.
more » « less- PAR ID:
- 10384697
- Publisher / Repository:
- Wiley Blackwell (John Wiley & Sons)
- Date Published:
- Journal Name:
- Small
- Volume:
- 18
- Issue:
- 19
- ISSN:
- 1613-6810
- Format(s):
- Medium: X
- Sponsoring Org:
- National Science Foundation
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