Now I am trying to purify polysaccharide from one strain of Pseudomonas aeruginosa. The medium is M63 with 0.4% arginine. I used 60% cold ethanol (final concentration) to purify the polysaccharide and get white precipitation. But the precipitation seems to lose function. And when I added cold ethanol to M63 medium(without bacterium, for control), I also get white precipitation.
So I wonder whether the white precipitation is arginine, because arginine is also Insoluble in ethanol. And any suggestion on my protocol on the polysaccharide purification. Should I change another medium?
Thank you very much!
Dear colleague,
In first you have to validate the chemical nature of your precipitate (sugar or not) with a colorimetric assay. If polysaccharide is in your supernatant with or without arginine precipitate you will detect sugars. If you have sugar, you can remove pollutant (arginine and other ) using tangential ultrafiltration as polysaccharides are high molecular weight compounds. If you have no sugar in your precipitate, try to cultivate your strain in another culture medium containing high amounts of carbon substrate.
Regards
Dear Colleague,
Dialysisshould be carried out against distilled water followed by microfiltration and finally precipitation in ethanol
Best regards
Dear Wang
I share you the experimental protocol of Bales et al. 2013 who have described how to purify polyssacharide from bacterial.
So you can also check another authors as Goncalves et al. 2007.
Thank
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3689685/
http://www.formatex.org/microbio/pdf/Pages450-457.pdf
Respected Imene Chentir
Kou Hayakawa
Hatem Majdoub Philippe MichaudCan you give me the protocol of purification and quantification capsule polysaccharide from gram negative bacteria sample like as Acinetobacter baumannii ? Thanks.
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