LACTB E.Coli, His Active

Recombinant DNA technology allows for the production of beta-lactamase in E. coli. This process involves inserting the gene encoding beta-lactamase into an expression vector, which is then introduced into E. coli cells. The cells are cultured, and the expression of the beta-lactamase gene is induced, leading to the production of the enzyme .

The pET-28a vector is commonly used for this purpose. It contains a T7 promoter, which is recognized by T7 RNA polymerase, and a lac operator, which allows for IPTG (isopropyl-β-D-thiogalactopyranoside)-inducible expression . The recombinant beta-lactamase is often fused with a His-tag (a sequence of histidine residues) at the N-terminus or C-terminus to facilitate purification using nickel affinity chromatography .

The recombinant beta-lactamase produced in E. coli typically contains 379 amino acids and has a molecular mass of approximately 41.8 kDa . The His-tagged version of the enzyme is purified using proprietary chromatographic techniques to achieve high purity levels (greater than 95% as determined by SDS-PAGE) .

The enzyme is supplied in a sterile, filtered, colorless solution, typically formulated in a buffer containing 20 mM Tris-HCl (pH 8.0) and 10% glycerol . For long-term storage, it is recommended to store the enzyme at -20°C with a carrier protein to prevent degradation .

Beta-lactamase enzymes, including the recombinant versions produced in E. coli, are crucial for studying bacterial resistance mechanisms and developing new antibiotics. They are also used in various research applications, including enzyme kinetics studies, structural biology, and drug discovery .