Tina ROSTINAWATI, Ami TJITRARESMI, Nyi Mekar SAPTARINI, Rahma ZISKA, Riezki AMALIA, Aisyah Sadira AZALIA, Harmony Anastasya NABABAN
Journal of Research in Pharmacy - 2026;30(2):467-481
L-Asparaginase (L-Asp) is an enzyme used in the treatment of acute lymphoblastic leukemia. However, current L-Asp therapies derived from bacterial sources often result in undesirable side effects. Plant-derived L-Asp, particularly from members of the Solanaceae and Fabaceae families, may offer a safer alternative. This study aimed to identify plant species from these families that exhibit L-Asp activity. Eleven plant species were initially selected. Crude protein was extracted using 1 M potassium phosphate buffer (pH 8.6) at a 1:2 (w/v) plant-to-buffer ratio. L-Asp activity was measured using the Nessler reaction, and protein content was determined. Subsequently, protein isolation techniques included salting out, dialysis, chromatography, Native PAGE, passive elution, enzyme activity and anticancer assay to reveal the active L-Asp candidate. Among the extracts, Archidendron pauciflorum seed extract demonstrated the most promising L-Asp activity, with a crude protein content of 237.08 µg/mL, L-Asp activity of 242.84 U/mL, and specific activity of 1.024 U/µg. The enzyme exhibited optimal activity at 40 dereceC and pH 8.8 in 0.5 M potassium phosphate buffer. The active L-Asp candidate had an approximate molecular weight of 70 kDa. The purified protein exhibited activity and specific activity of 240.74 U/mL and 0.129 U/µg, respectively. Cytotoxicity assays indicated that the active protein induced moderate cytotoxic effects on MCF-7 (breast cancer) and A549 (lung cancer) cell lines, with IC50 values of 6.43 and 8.07 U/mL, respectively. These findings suggest that plant-derived L-Asp, particularly from A. pauciflorum, holds potential as a safer alternative to microbial L-Asp in anticancer therapy. Furthermore, it highlights the potential for development into pharmaceutical or nutraceutical products while promoting the sustainable utilization of local biodiversity.
