Optimization of Three-Phase Partitioning system for Enhanced Recovery of L-Asparaginase from Escherichia Coli K12 Using Design of Experiment (DOE)

L-Asparaginase has been shown to possess anti-leukemic characteristic, mainly against Acute Lymphoblastic Leukaemia (ALL). L-Asparaginase produced by Escherichia coli K12 was purified by Three Phase Partitioning method of Iso-Octane in presence of ammonium sulphate. The factors like ammonium sulphate concentration, ratio of Iso-Octane to fermentation broth, temperature and pH which significantly affects the yield of desired enzyme during Three Phase Partitioning were selected for optimization. The purification of L-Asparaginase was optimized by use of Taguchi Design of Experiment methodology. L-16 array was selected for the purpose of optimization of TPP based purification. The interactions of different parameters were studied and they were used to formulate the optimized condition for the purification of L-Asparaginase. After the validation of result by performing the experiment under optimized condition there was 36.21% increase in the activity of the L-asparaginase, with 11 fold purification was achieved.