Recently, I am conducting a research about isolating Exopolysaccharides (EPS) produced by Lactobacillus bulgaricus. When I got the pellet of EPS after precipitaion with TCA and cold ethanol, the pellet cannot dissolved with deionized water. I want to dissolve it with deionized water because I want to do dialysis membrane against distilled water.
So, I am thinking about give some treatment such as give the higher pH or dissolve it in hot water. But, I am afraid those kind of treatment will change the structure of EPS, meanwhile I want to analyze the structure of EPS.
Does anyone have suggestion regarding to my research ?
Thank you.
Dear Arum Tiyas Suminar,
The Exopolysaccharides pellet precipitated from broth culture of lactic acid bacteria need to be dissolved in the 'deionized water' or 'double distilled water' only.
You must not go for any treatment to your EPS for its dissolution otherwise it may get affected.
Generally, the ethanol precipitated EPS is highly compact and its dissolution takes longer to dissolve in water.
You can keep your sample EPS in water in a shaker incubater (at 200 RPM) at 35 degree celcius to dissolve it.
Depending upon your sample size it will take time. I dissolved 15 gm of EPS in 30 mL of water; it took me 10-14 hours while it was kept in shaker incubator.
With regards,
Dr. A. K. Dubey
Maybe this article will help you:
New method for exopolysaccharide determination in culture broth using stirred ultrafiltration cells. Appl Microbiol Biotechnol. 2001; 57(3):401-6.
What if, if I dont conduct dialysis step for my EPS? I do not have dialysis bag for now.
I wanna conduct NMR analysis, do you think there will be any drawbacks?
Thanks
Best
decrease the concentration of EPS to better dissolve......
It is also better to spend more time for solving....
and use shaker incubator...
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