Production of L-asparaginase from Candida utilis by solid-state fermentation: a comprehensive assessment of its antiproliferative potential on glioblastoma cells

L-asparaginase (L-ASNase) is an enzyme that depletes asparagine, a key amino acid for cancer cell survival, producing aspartic acid and ammonia. Beyond its food industry applications, L-ASNase is a clinically important agent against acute lymphoblastic leukemia (ALL). In this study, L-ASNase was produced and purified from Candida utilis via solid-state fermentation. Optimization on wheat bran identified 2 mL inoculation volume, 60% moisture, and a 4-day fermentation period as the optimal conditions, yielding 172.5 U/mL activity. The purified enzyme was tested against glioblastoma (GBM) cell lines, showing IC50 values of 0.4 U/mL for U87MG and 1.8 U/mL for T98G, with minimal toxicity toward normal HaCaT cells. Apoptotic effects were confirmed by DAPI/F-actin and Giemsa staining, while wound healing and clonogenic assays revealed inhibition of cell migration and colony formation. RT-qPCR analysis demonstrated downregulation of the Survivin gene, a key survival regulator. These findings highlight L-ASNase's potent antiproliferative, anti-migratory, and pro-apoptotic effects, underscoring its potential as an adjuvant therapy for GBM.