Candidate Biopolymer Composite Membranes for Carbonic Anhydrase Immobilization in Enzymatic Direct Air Capture

Direct air capture (DAC) requires carbon capture interfaces that operate under highly dilute CO₂ conditions while minimizing thermal and chemical regeneration penalties. Carbonic anhydrase (CA) accelerates the reversible hydration of CO₂ to bicarbonate and is therefore a strong biocatalytic candidate for low-temperature CO₂ capture, but its implementation depends on candidate support materials that combine wet-state accessibility, chemical reactivity, mechanical processability and compatibility with membrane architectures. This study reports the preparation and screening of N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride/N-hydroxysuccinimide (EDC/NHS)-reactive biopolymer composite membranes for future carbonic anhydrase (CA) immobilization. Chitosan particles were precipitated with citrate or tripolyphosphate under high-shear homogenization and compared after lyophilization or convective drying. Chitosan-, shellac-, agarose- and cellulose-acetate-based films plasticized with glycerol and/or polyethylene glycol 400 (PEG-400) were then evaluated by optical microscopy, dry-state penetrometric puncture testing, qualitative EDC/NHS-reactivity mapping and Fourier-transform infrared spectroscopy (FTIR). Freshly precipitated chitosan particles showed dendrite-like high-surface morphologies, while lyophilization preserved porous flocculated aggregates and convective drying produced denser collapsed structures. Neat chitosan showed the highest dry-state puncture force (2.230 ± 0.173 N), whereas chitosan/shellac (0.377 ± 0.044 N) and agarose/chitosan/PEG-400 (0.386 ± 0.038 N) provided the strongest reactive-composite compromise between dry-state puncture resistance and EDC/NHS compatibility. The EDC/NHS reactivity map identified chitosan- and shellac-containing films as the chemically most relevant supports because they provide amine and/or carboxyl functionality, whereas agarose and cellulose acetate alone were not directly suitable for zero-length amidation. FTIR spectra confirmed polymer-specific functional signatures and EDC/NHS-associated changes in carbonyl, amide and C-O/C-O-C regions, especially in shellac- and chitosan-containing composites. The results identify chitosan/shellac as the lead candidate membrane and agarose/chitosan/PEG-400 as a hydration-rich comparator for subsequent carbonic anhydrase immobilization studies. This work should be interpreted as a first-stage materials-screening study of candidate membranes for enzyme immobilization.