Synergistic Soft‐Template Engineering of N‐Doped Hollow Mesoporous Carbon Spheres for High‐Efficiency Biocatalysis

ABSTRACT A novel emulsion‐induced interfacial assembly strategy was developed to synthesize hollow mesoporous carbon spheres (HM‐Cs) using the unique synergistic effect among the F127/P123 composite soft templates, pore‐expanding agent (TMB), and the carbon precursor (dopamine). The synthesis conditions were optimized by adjusting the P123/F127 mass ratio (19:1), carbonization temperature (550°C), and heating rate (2.5°C/min), resulting in HMC with well‐defined hollow cavities (150 nm) and radial mesopores (13‐20 nm). Characterizations via SEM, TEM, FTIR, TGA, XPS, and BET confirmed the material structural integrity, nitrogen doping, and high specific surface area (72.26 m 2 /g for HM‐C550). When used as carriers for Candida rugosa lipase (CRL) immobilization, HM‐C550 achieved a high protein loading capacity (127.58 mg/g). The immobilized lipase (HM‐C550@CRL) exhibited superior thermal stability and reusability compared to free CRL, retaining 34.08% of its activity after six hydrolysis cycles and 53.29% after six esterification cycles (cumulative activity 4.31 times that of the free enzyme). In the biosynthesis of phytosterol esters, optimal conditions (n‐hexane solvent, 40°C, 50 h, 20% enzyme dosage, 1:2 substrate molar ratio) yielded a maximum esterification rate of 76.99%. This work provides a controllable approach to fabricate functional carbon‐based carriers, advancing their application in efficient biocatalysis for functional lipid biosynthesis.