PURIFICATION AND CHARACTERIZATION OF L-ASPARAGINASE FROM A SOIL ISOLATE UNDER SOLID STATE FERMENTATION

Abstract

L-asparaginase has emerged as one of the most important clinically used enzymes as it exhibits chemotherapeutic potential in treatment of acute lymphoblastic leukimia and lymphosacroma. Soil microbial isolates were screened for potential producers of L-asparaginase using a phenol red indicator growth medium and the microbe producing the largest hydrolysis zone was selected. The isolate was characterized by biochemical tests and was found to belongs to Stenotrophomonas sp. The enzyme was partially purified by acetone precipitation with 73.32 % yield and a purification factor of 6.32 fold. Further purification includes gel filtration chromatoghraphy on Sephadex G-75 and C-25. The maximum enzyme activity was recorded at pH 7 and 35°C with a linear relationship concerning the increase in enzyme concentration. The effect of substrate concentration showed a progressive increase in the enzyme activity in a concentration dependent manner till it reaches a plateau where saturation was reached. The kinetics parameters (Km and Vmax ) of Stenotrophomonas sp L-asparaginase production were 96.71 mg/ml and 3333.33 umol/ml/min respectively.

Keywords: Characterization, L-Asparaginase., Purification, Stenotrophomonas sp


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