PURIFICATION AND BIOCHEMICAL CHARACTERIZATION OF RNA- DEPENDENT RNA POLYMERASE (RdRp) FROM FOOT AND MOUTH DISEASE VIRUS

Abstract

The RdRp protein (RNA dependent RNA polymerase) of the viruses plays an important role in the viral life cycle. This enzyme is considered an attractive target for viral preventive and treatment drugs. In this study, the recombinant RdRp protein of foot and mouth disease virus (FMDV) was purified and
examined biochemical activities. The recombinant RdRp protein with 98% purity were collected and expressed RNA polymerase activity with primer-dependent RNA synthesis. km and Vmax values are also calculated and km for UTP synthesis is 13.6 µM and Vmax is 42 µM / hour. Kcat for catalytic reaction is 0.28 min-1 and kobs for rate constant is 0.022 min-1. The inhibitor for enzyme activity was also tested and the IC50 for DMUT was 1.28 mM and 26.79 µM. Thus, the biochemical characteristics of FMDV RdRp has been identified, which is the basis for the screening of drug candidates in chemotherapy to prevent and treat foot and mouth disease.