Extraction And Purification Of Lipid A From Lipopolysaccharide Extracted From Proteus Mirabilis

Abstract

Current study included three parts: the first part is the isolation and identification ofOne case of Proteus mirabilis mixed with Pseudomonas and the other mixed with E.coli spp. And 1 case of Proteus mirabilis in pure form.from 50 samples were taken from infected ear of sheep. These isolates were identified by morphological examination and biochemical tests and vitak2 system. In the second part of this study, lipopolysaccharide (LPS) was extracted from Proteus mirabilis by using hot phenol method ,and after that crude extracted LPS purified by gel filtration chromatography using sephadex G-200 gel . 4 g. of partial purified LPS were prepared from cell of P. mirabiliscell , chemical analysis of LPS in 1 ml of crud and partial purified LPS showed that the carbohydrate percentages were (8, 17) % respectively ,while protein percentage were (14, 6) % respectively. The third part of this study Lipid A was separated from extracted lipopolysaccharide by A combination of mild-acid hydrolysis and solvent extractions (Bligh-Dyer) are used to liberate lipid A from covalently attached polysaccharide,830 mg of partial purified Lipid A were prepared from 1.5 g of lipopolysaccharide ..