Enterokinase Bovine
E. Coli.
Description
Enterokinase (rEK) Bovine Recombinant is the catalytic subunit of bovine enterokinase, which is expressed by E. Coli and purified to yield a high enzyme activity preparation. EK recognizes the sequence Asp-Asp-Asp-Asp-Lys and cleaves the peptide bond after the lysine residue. The enzyme can be used to cleave any fusion protein that carries this sequence. Recombinant Bovine Enterokinase is a single glycosylated polypeptide chain containing 235 amino acids and having an MW of ~28kDa.
Product Specs
E. Coli.
Product Science Overview
Enteropeptidase consists of two subunits:
- Heavy Chain: This subunit, with a molecular weight of approximately 115 kDa, anchors the enzyme to the intestinal membrane.
- Light Chain: The catalytic subunit, with a molecular weight of around 35 kDa, is responsible for the enzyme’s proteolytic activity .
The light chain of enteropeptidase recognizes and cleaves the peptide bond at the C-terminal end of the sequence Asp-Asp-Asp-Asp-Lys (D4K), which is essential for the activation of trypsinogen .
The recombinant production of the bovine enteropeptidase light chain has been a subject of extensive research due to its biotechnological applications. The recombinant form is produced in various expression systems, including Escherichia coli, Pichia pastoris, Aspergillus niger, and Saccharomyces cerevisiae . Among these, Escherichia coli is often preferred due to its high expression levels and cost-effectiveness .
However, the production of recombinant enteropeptidase light chain in Escherichia coli has faced challenges such as low yield and misfolding of the expressed protein. To address these issues, various strategies have been employed, including the use of fusion partners like thioredoxin and optimization of induction parameters .
The high specificity of enteropeptidase for the D4K sequence makes it a valuable tool in biotechnology. It is widely used for site-specific cleavage of fusion proteins, allowing for the production of target proteins without leaving unwanted amino acid residues . This property is particularly useful in the production of therapeutic proteins and industrial enzymes.
Recent studies have focused on improving the specificity and yield of recombinant enteropeptidase light chain. For instance, mutations at specific residues such as Tyr174 and Lys99 have been shown to enhance the enzyme’s specificity . Additionally, optimization of expression conditions, such as induction temperature and duration, has led to higher yields of active enzyme .
In conclusion, the recombinant production of bovine enteropeptidase light chain has significant potential in various biotechnological applications. Ongoing research aims to further improve the enzyme’s specificity and production efficiency, thereby expanding its utility in both laboratory and industrial settings.
