Enterokinase Bovine His

Enteropeptidase is a heterodimeric enzyme consisting of a heavy chain and a light chain linked by a disulfide bond. The heavy chain is responsible for anchoring the enzyme to the intestinal brush border, while the light chain contains the catalytic domain. The light chain of bovine enteropeptidase, which is often used in recombinant forms, consists of 235 amino acid residues .

Recombinant bovine enteropeptidase (rbEK) is produced using various expression systems, including E. coli. The recombinant form is typically engineered to include a His tag, which facilitates purification through nickel affinity chromatography. This His-tagged version is a single, non-glycosylated polypeptide chain containing 241 amino acids and has a molecular mass of approximately 28.0 kDa .

Recombinant enteropeptidase is widely used in biotechnology and research for its ability to specifically cleave fusion proteins at the recognition site. This specificity makes it an ideal tool for removing fusion tags from recombinant proteins, ensuring that the target protein remains intact and functional. The enzyme’s activity is stable across a wide range of pH levels (4.5–9.5) and temperatures (4–45°C), making it versatile for various experimental conditions .

The production of recombinant bovine enteropeptidase involves cloning the coding sequence of the light chain into an expression vector, followed by transformation into a suitable host, such as E. coli. The expressed protein is then purified using nickel affinity chromatography, leveraging the His tag for efficient isolation. The purified enzyme is typically formulated in a buffer containing Tris-HCl, NaCl, CaCl₂, and glycerol to maintain stability and activity .

Recombinant bovine enteropeptidase is stable for up to one year when stored at -20°C. It can also remain stable at 37°C for one week without significant loss of activity. However, repeated freeze-thaw cycles should be avoided to maintain enzyme integrity .