I would also like to known, what % of gel I should use in SDS-PAGE to find out the proper band of bacteriocin. Whether any specification for staining and De-staining...
Thanks in advance.
Bacteriocins are low molecular weight peptides.
Best method to purify bacteriocins from LAB is to use Tricine-SDS PAGE with upto 15% polyacrylamide gel concentration.
hope you will get desired results
gudluck!!
Some of bacteriocins can not be seen after Coomassie Blue staining so you can go for silver staining instead
Dear, you may see the following links for some insight.
Article Inhibitory effect of Lactobacillus acidophilus and Klebsiell...
Article Characterization of broad-spectrum klebocin produced by Kleb...
Grow a liquid culture of your bacteria into stationary phase. Then pellet the bacteria using centrifugation. Test the remaining supernatant for bacteriocin activity. If positive, run SDS PAGE gradient gel to separate proteins in the supernatant. Then run preparative gel to isolate.
We have obtaneid good results for the purfication of bacteriocins from LAB with the supernatant of the culture media by using XAD-16 resin followed by a solid-phase-extraction (C18 cartrige). In addition, it is important to highlight that the composition of the culture media may affect the purification process, usually MRS are quite complicated to use in this process. In the following publication and its references you can find more details.
Article Purification and characterization of antimicrobial peptides ...
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