This study investigates the fabrication and optimization of electrospun shellac fibers loaded with Senna alata leaf extract. The research employs a fractional factorial design and a Box–Behnken design to optimize electrospinning parameters that influence fiber morphology and extract entrapment efficiency. The optimal conditions were determined as a voltage of 24 kV, a solution feed rate of 0.8 mL/h, and a shellac–extract ratio of 38.5:3.8, resulting in nanofibers with a diameter of 306 nm, a low bead-to-fiber ratio of 0.29, and an extract entrapment efficiency of 96%. The fibers exhibited a biphasic release profile, with an initial burst release of 88% within the first hour followed by a sustained release exceeding 90% for 12 hours. The fibers demonstrated antimicrobial activity against various pathogens, suggesting potential applications in wound dressings and protective textiles. The SA leaf extract was prepared using ultrasound-assisted extraction with 95% ethanol, yielding 8.45% extract. Phytochemical analysis confirmed the presence of anthraquinones, flavonoids, terpenoids, saponins, tannins, and steroids, with rhein identified as a key compound. The electrospun fibers were characterized using SEM, PXRD, DSC, and FTIR, revealing a nanosized morphology with minimal bead formation. The entrapment efficiency of rhein was determined using HPLC-DAD, with values ranging from 58.88% to 105.42%. The in vitro release study showed a biphasic release profile, with the fibers exhibiting sustained release over 12 hours. The antimicrobial activity of the fibers was assessed using a time–kill kinetics assay, demonstrating effective inhibition of bacterial growth. The results indicate that electrospun shellac fibers loaded with SA leaf extract have potential for use in wound healing and antimicrobial applications.This study investigates the fabrication and optimization of electrospun shellac fibers loaded with Senna alata leaf extract. The research employs a fractional factorial design and a Box–Behnken design to optimize electrospinning parameters that influence fiber morphology and extract entrapment efficiency. The optimal conditions were determined as a voltage of 24 kV, a solution feed rate of 0.8 mL/h, and a shellac–extract ratio of 38.5:3.8, resulting in nanofibers with a diameter of 306 nm, a low bead-to-fiber ratio of 0.29, and an extract entrapment efficiency of 96%. The fibers exhibited a biphasic release profile, with an initial burst release of 88% within the first hour followed by a sustained release exceeding 90% for 12 hours. The fibers demonstrated antimicrobial activity against various pathogens, suggesting potential applications in wound dressings and protective textiles. The SA leaf extract was prepared using ultrasound-assisted extraction with 95% ethanol, yielding 8.45% extract. Phytochemical analysis confirmed the presence of anthraquinones, flavonoids, terpenoids, saponins, tannins, and steroids, with rhein identified as a key compound. The electrospun fibers were characterized using SEM, PXRD, DSC, and FTIR, revealing a nanosized morphology with minimal bead formation. The entrapment efficiency of rhein was determined using HPLC-DAD, with values ranging from 58.88% to 105.42%. The in vitro release study showed a biphasic release profile, with the fibers exhibiting sustained release over 12 hours. The antimicrobial activity of the fibers was assessed using a time–kill kinetics assay, demonstrating effective inhibition of bacterial growth. The results indicate that electrospun shellac fibers loaded with SA leaf extract have potential for use in wound healing and antimicrobial applications.