I am trying to extract Bacillus bacteriocins, do I stop stirring when all the ammonium sulfate has been dissolved or prolonged stirring helps in extraction before moving sample to 40C overnight.
Dear Nelisiwe,
Solubility of ammonium sulfate is completely differ from bacteriocin precipitation. After calculation of required solid ammonium sulfate, you must add ammonium sulfate slowly to supernatant of the culture medium at 4°C with gentle magnetic stirring. After solubilization of ammonium sulfate, bacteriocin precipitation should continue for 24 hours at 4°C with gentle magnetic stirring. Then, precipitate should be removed by centrifugation at 4°C. This answer is according to my experience working with nisin.
Thank you so much it worked. I am extracting Bacillus and realized the bacteriocin pellets float with extensive stirring so after stirring to dissolve the ammonium sulfate for about 2 hours I leave it at 4°C without stirring overnight. Your response helped.
Dear say-yed... could you explain me on how to calculate the ammonium sulfate concentration need for bacteriocin precipitation...
Dear Asbar Tanjung,
You have 3 options.
1.Calculating needed amount of ammonium sulfate
2.Using predefined tables for specific volumes
I recommend studying the attached article.
3.Using online calculators such as the following hyperlink:
https://www.encorbio.com/protocols/AM-SO4.htm
Do not forget that temperature affects the needed amount of ammonium sulfate.
Dear Say-yed.... alot of thanks for these answer, these very help full for my research stagnancy so far..... I'll try... could you recommend other articles or protocol... I cant acces the attached file
Dear Asbar Tanjung,
If you have not access to the article, please send me your email to:
I will send you it by email.
Dear Say-Yed Hesameddin, Okey, Thanks before.. I'v sent you an email...
Dear Say-yed Hesameddin Tafreshi, I've got these attached files... Thanks again for these files... these will be very help full for me and my research