Extraction, Purification and Determining Molecular Weight of Exopolysaccharide Produced by Pseudomonas Spp. Isolated From Urine

Abstract

Background: Microbial exopolysaccharides (EPSs) are well-recognized biotechnology products that are generated by different genera of bacteria and yeasts. pharmaceutical, as well as in the medical domain. As a model for the study of EPS-produced microorganism , Pseudomonas spp. synthesizes significant amounts of (EPSs). In this study, we aim to Extract and purify (EPSs) produced by Pseudomonas isolated from urine samples for their molecular characterization, knowing their composition, and investigating their molecular weights. Methods: Exopolysaccharides were extracted from (43) clinical isolates of Pseudomonas spp. using hot phenol method, (EPSs)were obtained from the aqueous phase and purified using Sephadex G200 gel and MW was determined by gel filtration technique. Result: It was found that the percentage of carbohydrates in the purified extract was 76.96% and proteins was 20.84%, while in the crude extract it was 71.32 and 30.76% for carbohydrates and proteins respectively, exopolysaccharides were detected by the Limulus amebocyte lysate (LAL) reagent to detect the biological effectiveness, while MW of the purified extract was 126.993 kDa Conclusions: Pseudomonas spp. produces a large amount of EPS and the results of the current research found that the percentage of proteins and nucleic acids contaminating the extract decreases when the extract is re-purified, also found that the MW of EPS extract was 126.993 kDa. Further study is required for the purpose of the possibility of providing good conditions for the production of larger quantities of EPSs from Pseudomonasspp. to the extent that they can be used in industry, the medical field, and others.