I am working on the bacterial exopolysaccharides, I need to check purity of the polysaccharide. Can anybody tell me what are the methods have been used for the above?
Dear Dr. Saranya K.,
The bacterial exopolysaccharides (EPS) are generally contaminated with the broth proteins. You may like to estimate protein in your EPS sample using bradford or lowry method. If there is any protein contamination then you need to again extract the EPS (dissolved in double distilled water) with ice cold ethanol. Subsequently, the EPS may alos be extracted using a volume of chloroform and iso-amyl alcohol (24:1)equal to volume of EPS, followed by re-precipitation using ethanol. This step involving chloroform and iso-amyl alcohol can be repeated 3-4 times to remove protein contaminants.
The other non-proteinaceous contaminants can be removed from the EPS solution using 1kDa dailysis bag.
After dialysis, the contamination in EPS would be minimum to zero.
Best Wishes,
Dr. A. K. Dubey, PhD
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