Optimization of expression, purification and handling anti bacteria feature protein of bovine neutrophil B-defensing BNBD2

Defensins are one of the largest families of antimicrobial peptides and, because of their activity against bacteria, fungi, and many coated and uncoated viruses are very profitable as new antibiotics. This study was done to evaluate the optimization of expression, purification and handling antibacterial features of BNBD2 protein.

In this experimental laboratory study, E. coli BNBD2 (DE3) carrying pET-32a(+) vector in which BNBD2 was homogenized was used. BNBD2 expression was evaluated through changes in cell density, growth temperature, the duration of induction using vertical electrophoresis (SDS-PAGE) and Bradford test both qualitatively and quantitatively. The purification of recombinant protein was done by chemical method of fission in the position of formic acid and passing Centricon, and anti-bacterial effect of the purified protein on some gram negative and gram positive bacteria was examined

By induction start in optical absorption of 0.8 in wave length of 600 nm and inducing IPTG in concentration of 1 milimolar, growth temperature of 300C, and duration of 4h after induction, the highest expression was obtained in Luria-Bertani culture medium. The recombinant protein was purified by fission in formic acid and passing centricon. Western blot test results indicated that the recombinant protein was specifically connected to mouse anti-(His) 6 peroxidase antibody. The formation of inhibition zone in culture media of Hinton agar containing culture swab of the studied bacteria indicated this protein’s antibacterial property.

BNBD2 protein has antibacterial effects and can be expressed in E. coli and this protein has antibacterial effects.