L-Asparaginase (E.C. 3.5.1.1) is an amidohydrolase enzyme which catalyze L-asparagine to L-aspartate and generate ammonia. This enzyme has an important application on medicine and food processing aid. In this research, L-asparaginase from food grade microorganism, Arthrospira platensis was cloned and expressed in Escherichia coli system. The expression of cloned gene resulted in the inclusion body form. It was found that deduced protein was located and accumulated in pellet fraction, inclusion bodies. Hence, to achieve active form of the enzyme, it need to be correctly folded with steps as follow solubilization and refolding. Here, we report the recovering activity of the enzyme throughout our refolding process. Several denaturant agents were used to completely denature a protein from inclusion body. Urea, triton-X 100, and sarkosyl are able to denature the protein. The refolding process was conducted by dilution and dialysis methods. After experienced refolding process, L-asparaginase activity was detected. The highest activity was achieved from dialysis methods (64 U). We, therefore, reported the activity in the refolded enzyme even though the process needs to be optimized.

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