Acute lymphocytic leukemia is a common leukemia characterized by frequent infections and anemia. Thousands of new cases are diagnosed each year worldwide. L-Asparaginase (E.C.3.5.1.1), also known as L-asparagine amino hydrolase, is a potential anti-tumor enzyme that catalyses the hydrolysis of L-asparagine into L-aspartic acid and ammonia. L-Asparaginase production was investigated in the isolated filamentous fungi on sesame cake using solid state fermentation (SSF). L-Asparaginase production using isolated Aspergillus niger grown on sesame cake has been optimized in a column bioreactor using a statistically-based method. The 23  BHH   design having   eight   different   fermentation   conditions   was   applied to evaluate their significance on L-Asparaginase activity, where the three independent variables evaluated were  Aeration,  Thickness  of  Solid  Substrate  Bed  and  Fermentation  Temperature. Aeration and Temperature were identified to be important variables and had a positive effect on  responses,  however,  the  thickness  of  solid  substrate  had  an  insignificant  effect  on  L-Asparaginase activity  within the tested range. The optimum fermentation condition for L-Asparaginase activity consisted of high aeration (0.4 vvm), deeper thickness of bed (22 cm) and high fermentation temperature (32°C), respectively. Under optimum levels, a maximum L-Asparaginase activity of 344.2132 IU was predicted which is slightly less than the activity at batch level.