Extraction and Purification of Lipopolysaccharide from Escherichia coli (local strain) and study its pyrogenic activity
AbstractThe Escherichia coli (E. coli) bacteria belong to the Enterobacteriaceae family can cause wide ranges of infections including enteric, extra enteric or opportunistic infection especially in young animals. E. coli outer membrane composed of phospholipid and lipopolysaccharide which reflect the inner and outer layer of the membrane, the main components of lipopolysaccharide LPS are lipid A, core oligosaccharide, and O- antigen. Immunity of LPS was clearly understood in the last 20 years when discovered signaling response in immune cell system, LPS acting as potent activators of immune system especially innate immunity and as a result produce a large variety of the adverse, unfavorable physiological response leading to injury of tissue and endotoxic shock and lastly death.
In this study, we tried to extract and purify the LPS from E.coli local strain and determine the molecular weight, purity, and pyrogenic effect of the product and compare with stander E.coli O55:B5 LPS, the result shows that the hot phenol method with enzymatic treatment give highly pure LPS with high yield reach to 242.4 mg, staining the SDS page gel with Coomassie blue and silver nitrate uncover the high purification of the extracted LPS (ELPS) with no protein contamination, with molecular weight range between 15-23 kDa, HPLC test reveals that purity of ELPS was 100 % compared with standard LPS. The rabbits' pyrogen test confirmed that the biological activity of ELPS. In conclusion, the method of extraction used in this research was valid and give highly pure LPS with enduring substantial pyrogenic activity.
- Article View: 19
- PDF Download: 0