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Updated: Sep 17, 2025
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Immobilization of Multi-biocatalysts in Alginate Beads for Cofactor Regeneration and Improved Reusability
Published on: April 22, 2016
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Coprecipitated Enzyme-Encapsulated Covalent Organic Frameworks for Biocatalysis
Satyadip Paul1, Mani Gupta2, Shayan Karak1
1Department of Chemical Sciences, Indian Institute of Science Education and Research, Mohanpur, Kolkata 741246, India.
Journal of the American Chemical Society
|June 27, 2025
Summary
We developed a one-pot synthesis to encapsulate enzymes in covalent organic frameworks (COFs), enhancing their stability and reusability. This method improves enzyme performance under harsh conditions, enabling broader biocatalysis applications.
Area of Science:
- Biocatalysis
- Materials Science
- Nanotechnology
Background:
- Enzymes are efficient biocatalysts but exhibit limited stability under harsh industrial conditions.
- Developing robust enzyme immobilization strategies is crucial for expanding their practical applications.
- Covalent organic frameworks (COFs) offer a promising platform for biomolecule encapsulation due to their tunable structures and high surface areas.
Purpose of the Study:
- To develop a facile aqueous synthesis for enzyme-encapsulated COFs.
- To investigate the interactions between encapsulated enzymes and the COF matrix.
- To evaluate the enhanced stability and reusability of encapsulated enzymes.
Main Methods:
- One-pot aqueous synthesis of TpAzo COFs encapsulating various enzymes, including beta-glucosidase (BGL) and alkaline phosphatase (ALP).
- Solid-state 2D NMR correlation spectroscopy to probe enzyme-COF interactions at the molecular level.
- Scattering-type scanning near-field optical microscopy (s-SNOM) and nanoscale Fourier-transform infrared spectroscopy (nanoFTIR) for validation.
- Assays to measure enzyme activity, stability under denaturing conditions (SDS), and recyclability.
Main Results:
- Successful encapsulation of multiple enzymes and proteins within the TpAzo COF.
- Direct evidence of molecular interactions between enzymes and the COF backbone, confirming structural integrity.
- Encapsulated BGL and ALP retained high catalytic activity and were recyclable for up to ten cycles.
- COF encapsulation significantly enhanced BGL stability in 1-15% SDS solutions, mitigating denaturation.
Conclusions:
- A robust one-pot aqueous synthesis strategy for enzyme-COF composites was established.
- Enzyme-COF interactions are key to enhancing enzyme stability and reusability.
- This approach offers a viable method for creating stable, reusable biocatalysts for demanding applications.
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