Stability and Kinetic Study of Immobilized Carbonic anhydrase into PVDF Membrane

Siti Nadia  Abdullah; Fazlena Hamzah; Nursyuhani Che  Husain; Harumi Veny; Miradatul Najwa Mohd Rodhi; Nur Atikah Mohidem; Dessy Ariyanti

doi:10.22146/ajche.17764

Siti Nadia Abdullah Faculty of Chemical Engineering, Universiti Teknologi MARA Cawangan Johor, Pasir Gudang Campus, Jalan Purnama, Bandar Seri Alam, 81750 Masai, Johor, Malaysia
Fazlena Hamzah Biocatalysis and Biobased Material Technology Research Group, Faculty of Chemical Engineering, Universiti Teknologi MARA, UiTM Shah Alam, 40450 Shah Alam, Selangor, Malaysia
Nursyuhani Che Husain Biocatalysis and Biobased Material Technology Research Group, Faculty of Chemical Engineering, Universiti Teknologi MARA, UiTM Shah Alam, 40450 Shah Alam, Selangor, Malaysia
Harumi Veny Biocatalysis and Biobased Material Technology Research Group, Faculty of Chemical Engineering, Universiti Teknologi MARA, UiTM Shah Alam, 40450 Shah Alam, Selangor, Malaysia
Miradatul Najwa Mohd Rodhi Biocatalysis and Biobased Material Technology Research Group, Faculty of Chemical Engineering, Universiti Teknologi MARA, UiTM Shah Alam, 40450 Shah Alam, Selangor, Malaysia
Nur Atikah Mohidem i-CATS University College Faculty of Engineering, Jalan Stampin Timur, 93350 Kuching, Sarawak, Malaysia
Dessy Ariyanti Department of Chemical Engineering, Faculty of Engineering, Universitas Diponegoro, Jl. Prof. Soedarto, Tembalang, Kec. Tembalang, Kota Semarang, Jawa Tengah 50275, Indonesia

DOI: https://doi.org/10.22146/ajche.17764

Keywords: Carbonic anhydrase, Enzyme Kinetics, Enzyme Stability, Immobilization, PVDF Membrane

Abstract

Carbonic anhydrase (CA) is a highly efficient biocatalyst for accelerating CO₂ hydration and has attracted significant interest for enzyme-assisted carbon mineralization and post-combustion CO₂ capture. However, the practical deployment of CA is hindered by its limited stability and non-recyclability under industrially relevant conditions. In this study, CA was immobilized onto a hydrophobic poly(vinylidene fluoride) (PVDF) membrane via glutaraldehyde-mediated covalent crosslinking to develop a robust enzymatic membrane platform for CO₂ mineralization applications. The immobilized enzyme showed greater enhanced thermal and pH stability than free CA, demonstrating improved resilience under conditions relevant to mineral carbonation processes. Enzymatic kinetics were systematically evaluated using p-nitrophenyl acetate as a model substrate, revealing a low apparent Michaelis constant (K_m = 7.45 mmol L⁻¹) and a maximum reaction rate (V_m = 0.76 µmol min⁻¹), indicative of strong enzyme–substrate affinity within the membrane-confined microenvironment. Scanning electron microscopy confirmed homogeneous enzyme distribution and stable attachment within the PVDF pore structure. While employing a conventional immobilization strategy, this work provides quantitative insight into the stability–kinetics relationship of membrane-immobilized CA. It establishes a baseline membrane architecture for future development of advanced enzyme-assisted CO₂ mineralization and membrane contactor systems.

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