The Effect of Additional Indonesian Bioactive Compounds on Activity of Mutant Catalase-Peroxidase

• Main Author: Rahman Tara, Fedi
• Format: Final Project
• Language: Indonesia
• Subjects: Kimia
• Online Access: https://digilib.itb.ac.id/gdl/view/41306
• Institution: Institut Teknologi Bandung

Tuberculosis (TB) is an infectious disease caused by Mycobacterium tuberculosis (M. tuberculosis). Since 1952, TB have been treated using isoniazid (INH). INH is a prodrug that is activated to a TB-drug by the catalase-peroxidase enzyme (KatG) of M. tuberculosis. The effectiveness of INH as a tuberculosis drug reduced extensively because of genetic mutations in M. tuberculosis, particularly in a gene encoding KatG enzyme, called as the katG gene. Continuous evolutions of bacterial resistance to widely used antibiotics have triggered the need of discovering of new and more effective antimicrobial drugs. Indonesia has abundant active natural compounds, such as compound-1 and compound-2. The aim of this study is to determine the effect of additional compound-1 and compound-2 on the activity of the Ile518Thr mutant KatG in activating INH. The Ile518Thr mutant protein was purified by the affinity chromatography method using Co-NTA resin resulted in three dominant bands at 80, 45, and 35 kDa, respectively in an SDS-PAGE electropherogram. A band at 80 kDa was related to KatG while the other bands probably were due to degraded KatG proteins as a result of protease activity from E.coli, as the host cells. The specific activity of the Ile518Thr mutant decreased by 68% compared to that of wildtype KatG. Interestingly, addition of compound-2 to the Ile518Thr mutant KatG restored the activity of the mutant KatG in activating INH by 77%. Meanwhile, the addition of compound-1 to the same mutant KatG, showed no significant changes in the activity of Ile518Thr KatG. Based on these results, it is likely to suggest that the compound-2 plays a role as an effector of KatG in activating INH as an antituberculosis drug.