REFOLDING OF PROTEIN, NIES39_A07830 FROM Arthrospira platensis AND ITS L-ASPARAGINASE ACTIVITY

Asep A. Prihanto (Corresponding Author)
(Dept. Fish Processing Technology, Faculty of Fisheries and Marine Science, Brawijaya University, Indonesia/ asep_awa@ub.ac.id)
(Center for Coastal and Marine Studies, Brawijaya University, Indonesia/ asep_awa@ub.ac.id)

Happy Nursyam
(Dept. Fish Processing Technology, Faculty of Fisheries and Marine Science, Brawijaya University, Indonesia/happy_nursyam@yahoo.com)

Abstract
L-Asparaginase (E.C. 3.5.1.1) is an amidohydrolase enzyme which catalyze L-asparagine to L-aspartate and ammonia. This enzyme has an important applications 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 cloned gene expressed in
the inclusion body form. It was found that deduced protein was located and accumulated in the pellet fraction, as inclusion bodies. Hence, to achieve an active form of the enzyme, it was needed 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.

Keywords: Refolding, inclusion body, L-asparaginase, food grade, Arthrospira platensis,

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