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Nitrilotriacetic Amine-Functionalized Polymeric Core-Shell Nanoparticles as Enzyme Immobilization Supports
Dominic Keller1,2, Ana Beloqui1,2, Mónica Martínez-Martínez3
1Institute of Toxicology and Genetics, Karlsruhe Institute of Technology (KIT) , Hermann-von-Helmholtz-Platz 1, 76344 Eggenstein-Leopoldshafen, Germany.
Biomacromolecules
|July 22, 2017
Summary
Nitrilotriacetic amine-functionalized nanoparticles were created using polymerization-induced self-assembly for enzyme immobilization. These novel nanobiocatalysts efficiently immobilize His-tagged enzymes, maintaining their catalytic activity.
Area of Science:
- Polymer Chemistry
- Nanotechnology
- Biocatalysis
Background:
- Enzyme immobilization is crucial for biocatalyst reusability and stability.
- Developing efficient and well-defined nanoparticle supports is essential for advanced immobilization strategies.
- Nitrilotriacetic amine (NTA) functionalization offers a specific binding site for polyhistidine-tagged proteins.
Purpose of the Study:
- To synthesize nitrilotriacetic amine (NTA)-functionalized nanoparticles via aqueous polymerization-induced self-assembly (PISA).
- To evaluate these nanoparticles as supports for immobilizing polyhistidine-tagged (His-tagged) enzymes.
- To create catalytically active nanobiocatalysts with enhanced properties.
Main Methods:
- Synthesis of a novel nitroxide-mediated polymerization initiator with a protected NTA moiety.
- Preparation of amphiphilic block copolymer nanoparticles using PISA and nanoprecipitation.
- Characterization of nanoparticles using size-exclusion chromatography (SEC), electrospray ionization mass spectrometry (ESI-MS), and dynamic light scattering (DLS).
- Assessment of NTA deprotection and nickel ion complexation using DLS and ICP-OES/MS.
- Immobilization of His-tagged horseradish peroxidase and ester hydrolase.
Main Results:
- Well-defined, end-functional vinyl polymers were produced using the novel initiator.
- PISA yielded spherical nanoparticles with higher definition compared to nanoprecipitation.
- Successful NTA deprotection and nickel ion complexation were confirmed.
- Catalytically active nanobiocatalysts were successfully prepared by immobilizing His-tagged enzymes.
Conclusions:
- Aqueous PISA is an effective method for creating well-defined NTA-functionalized nanoparticles.
- These nanoparticles serve as efficient supports for immobilizing His-tagged enzymes.
- The resulting nanobiocatalysts retain significant enzymatic activity, demonstrating their potential for practical applications.
