I'm extracting proteins from orange tomato fruit. After the step of centrifuge the extract forms the gel so I do not have any pellet where proteins are normally found. Probably it is due to high content of pectin in orange fruit. (In green fruit I don't have this problem). What should I do to avoid this gelation?
The pH is 8.
I think we found a way to solve the problem. The addition of EGTA at 0,5 M to the extraction buffer stops the gelation process. It is chelating calcium ions necessary for pectin gelation.
You can try digest the pellet with pectate lyase or polygalacturonase afterwards to release the protein
Using Pectinase enzyme is the best possible solution for your problem.
pectin gelation is temperature and pH dependent, centrifuge at 25 or 30 oC
I had the same problem with apple fruit extracts. Divalent cation chelators will help. Another simple way is to dilute your axstracts a bit.
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