Truncation of C-Terminal Intrinsically Disordered Region of Mycobacterial Rv1915 Facilitates Production of "Difficult-to-Purify" Recombinant Drug Target.

ABSTRACT: Availability of purified drug target is a prerequisite for its structural and functional characterization. However, aggregation of recombinant protein as inclusion bodies (IBs) is a common problem during the large scale production of overexpressed protein in heterologous host. Such proteins can be recovered from IB pool using some mild solubilizing agents such as low concentration of denaturants or detergents, alcohols and osmolytes. This study reports optimization of solubilization buffer for recovery of soluble and biologically active recombinant mycobacterial Rv1915/ICL2a from IBs. Even though the target protein could be solubilized successfully with mild agents (sarcosine and βME) without using denaturants, it failed to bind on Ni-NTA resin. The usual factors such as loss of His6-tag d

SUBMITTER: Antil M 

PROVIDER: S-EPMC7273500 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature