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In this paper, sulfonic groups functionalized annealed bio-based carbon microspheres loaded polytetrafluoroethylene (A-BCMSs-SO3H@PTFE) fibers with high activity, high stability, and easy regeneration were successfully fabricated by a simple method using low-cost raw materials. The characterization results showed that the annealed biomass carbon microspheres derived from waste Camellia oleifera shells were evenly distributed on the polytetrafluoroethylene fibers and the sulfonic groups can be successfully loaded on the surface of annealed biomass carbon microspheres by room temperature sulfonation. Subsequently, the as-prepared A-BCMSs-SO3H@PTFE fibers were applied to the acid-catalyzed synthesis of liquid biofuel 5-ethoxymethylfurfural. The catalytic experiment results indicated that the annealing temperature and time during catalyst preparation have a significant effect on the activity and selectivity of A-BCMSs-SO3H@PTFE fibers. The results of catalytic reaction kinetics showed that the yield of 5-ethoxymethylfurfural can reach more than 60％after 72 h of acid-catalyzed reaction. The stability test showed that the as-prepared A-BCMSs-SO3H@PTFE fibers still maintained a stable acid catalytic activity after four recycles.