Author/Authors :
ROUHANI, Maryam Department of Nano-Biotechnology - New Technologies Research Group - Pasteur Institute of Iran, Tehran, Iran , VALIZADEH, Vahideh Department of Nano-Biotechnology - New Technologies Research Group - Pasteur Institute of Iran, Tehran, Iran , MOLASALEHI, Sara Department of Nano-Biotechnology - New Technologies Research Group - Pasteur Institute of Iran, Tehran, Iran , NO-ROUZIAN, Dariush Department of Nano-Biotechnology - New Technologies Research Group - Pasteur Institute of Iran, Tehran, Iran
Abstract :
Background: Serratiopeptidase is a bacterial metalloprotease, which is useful for the treatment of pain and inflammation. It breaks down fibrin, thins the fluids formed during inflammation and acts as an anti-biofilm agent. Because of medicinally important role of the enzyme, we aimed to study the cloning and the expression optimization of serratiopeptidase.
Methods: The heat-stable serratiopeptidase (5d7w) was selected as the template. Cloning into pET28a expression vector was performed and confirmed by colony PCR and double restriction enzyme digestion. The recombinant protein was expressed in Esherichia coli BL21 and confirmed by SDS-PAGE and Western blot analysis. Different parameters such as expression vector, culture media, post-induction incubation temperature, inducer concentration, and post-induction incubation time were altered to obtain the highest amount of the recombinant protein.
