Production of biochemically significant yields of recombinant mycobacterium smegmatis rhomboid protease i

Abstract

The rhomboids are a recently discovered, well-conserved family of serine proteases. well-conserved family of serine proteases has been identified as a viable novel drug target in Mycobacterium TB and this could be very vital for the management of this disease as the future progresses. However, the research into it being consistently hampered due to the requirement of significant amounts of protein required which is difficult in the case of intramembrane proteins like rhomboids. In this excerpt therefore, we tasked ourselves with finding the most suitable conditions that can favour the production of significant amounts of rhomboid protease 1 in Mycobacterium smegmatis including the concentration of the inducing agent (IPTG), induction time and temperature. We found these to be 3.5mM, 12 hrs and 300C respectively. We do however think that this procedure can be further optimised be trying out other values for these particular conditions or even exploiting more complex techniques like codon optimisation.