Improved recombinant expression of soluble cathepsin B and L in Escherichia coli.

ABSTRACT: Cysteine cathepsins such as cathepsin B and L play an important role in numerous diseases like acute pancreatitis or SARS-CoV-2 and therefore have high potential for the development of new therapeutics. To be able to screen for potent and selective inhibitors sufficient amounts of protein are required. Here, we present an easy and efficient protocol for the recombinant expression of soluble and active murine cathepsin B and L. For this, we used the strain E. coli SHuffle® T7 Express which is capable of forming disulfide bridges in the cytoplasm. The enzymes were purified by immobilized nickel ion-affinity chromatography. Using different constructs and media, expression levels were significantly improved and expression yields of 80 ± 2 mg L^-1 for procathepsin B, which is 16-fold

SUBMITTER: Moller C 

PROVIDER: S-EPMC11649755 | biostudies-literature | 2024 Dec

REPOSITORIES: biostudies-literature