Characterization of adenosine deaminase from Mycobacterium tuberculosis

Author: Will Osae
Major: Biochemistry
Approved: Spring 2018
Status: Completed

Adenosine deaminase (ADA) is an enzyme that participates in the metabolism of adenine nucleotides. It catalyzes the irreversible deamination of 2’- deoxyadenosine into 2’-deoxyinosine and that of adenosine into inosine. In this paper, we discuss our study of the ADA gene from Mycobacterium tuberculosis (MTB), the causative agent of tuberculosis (TB). Very little study of this gene has been performed despite a recent rise in interest of measuring ADA levels in blood serum as an effective diagnostic tool for tuberculosis and other ailments. In this study, competent E. coli cells were transformed with the ADA gene from MTB, and the cells were induced to overexpress the ADA protein afterwards. Maltose-binding protein (MBP) was used as an affinity tag for purification of the ADA protein via amylose resin chromatography. The purified enzyme was assayed for activity and an estimated Km of 0.279 mM was obtained.