Optimization of recombinant vanadium-dependent haloperoxidase production from gracilaria changii

Vanadium-dependent haloperoxidases (VHPOs) belong to one of the three classes of \nhaloperoxidases which can oxidize halogens including chloride (Cl-), bromide (Br-), \nand iodide (I-). Chemical halogenation of organic compounds typically requires harsh \nconditions. Hence, biohalogenation has gained increasing research interest. As \nvanadium-dependent haloperoxidases are enzymes that catalyze halogenation of \norganic compounds, they are very valuable due to their potential applications in \nvarious industries as well as their stability and tolerance for different conditions. This \nstudy aims to optimize the recombinant protein expression conditions, for potentially large scale production of vanadium-dependent bromoperoxidase 2 (GcVBPO2) that \nwas previously isolated from Gracilaria changii. GcVBPO2 sequence in pET32a(+) \nwas transformed into expression host Escherichia coli BL21(DE3) pLySs and induced \nat various temperatures. GcVBPO2 was found to be soluble when induced with \n0.5mM Isopropyl β-D-1-thiogalactopyranoside (IPTG) for 16 hours in Luria-Bertani \n(LB) broth culture at 20°C. Purification of GcVBPO2 was performed using His-tag \npurification, and subsequently analysed with Western blot.