Kinetic Study of Plasmodium falciparum Orotate Phosphoribosyl Transferase in the Presence of Activator and Inhibitor
การศึกษาจลนศาสตร์ของเอนไซม์ออโรเทต ฟอสฟอไรโบซิลทรานเฟอเรสของเชื้อพลาสโมเดียม ฟัลซิพารัมในสภาวะที่มีตัวกระตุ้นและตัวยับยั้ง
Keywords:
Plasmodium falciparum , orotate phosphoribosyl transferase , activation, inhibition, metal ionAbstract
Objective: To find the chemicals which have the stimulating or inhibitory properties on orotate phosphoribosyl transferase enzymes (OPRT) of Plasmodium falciparum. Method: construction of PfOPRT gene was ligated in pQE 30Xa plasmid and transfer into E. coli M15 and prove the construct recombinant plasmid. After it was proved to be the desired gene, the enzyme was expressed and purified. The performance was tested and metal ions were tested for activation and Orotidine 5'-Monophosphate (OMP) reaction products for inhibition. Result: The effective cofactor of OPRT is Mg2 + with Vmax = 64.25 µmol/min and Km = 58.82 µM and the enzyme activity without a cofactor is equal to zero. The cofactor was converted to Zn2 +, the OPRT activity was reduced was show Vmax = 9.35 µmol/min and Km= 136.24 µM. A reverse inhibition test was performed using the product obtained from the reaction OMP, resulting in a competitive inhibition of OPRT.
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