Abstract
l-asparaginase (L-ASNase) has garnered considerable interest in lymphoma therapy due to its mechanism of action, which involves depleting the availability of circulating l-asparagine, a crucial nutrient for malignant cells. Given the notable side effects of commercially available bacterial L-ASNase and the existing challenges in large-scale industrial production, this study focused on identifying and optimizing novel fungal sources for the enzyme. The research methodology involved initial strain screening, development and optimization of a bioreactor system, and subsequent evaluation of enzyme production within a bioprocess. A comprehensive screening of various fungal strains revealed that Rhizopus sp. UPX271019 exhibited the highest specific activity, indicating its significant potential for therapeutic and industrial applications. The study investigated the performance of various bioreactor configurations. A fixed-bed airlift hybrid bioreactor (FB-ALR), operated with an aeration rate of 0.75 vvm, demonstrated superior hydrodynamic and mass transfer performance. A critical observation was the significant role of biofilm formation by Rhizopus sp. UPX271019 within the FB-ALR, which was instrumental in achieving substantial L-ASNase production. The bioprocess yielded an average enzyme activity of 2475 ± 701 U, highlighting the system's effectiveness. These results suggest that the FB-ALR operating with such a strain grown as biofilm is a highly suitable system for future industrial scale-up.