My current ongoing research is in the isolation of antimicrobial compounds from actinomycetes and fungi. My team used various microorganisms and studied antimicrobial activity on solid as well as liquid media. I would like to know what method of purification we should apply so that we can have minimum loss and maximum activity.
It really depends which compound showed the activity? Is it a proteine? another compound? How big? How polar?
each would need a different purification protocol
Is it totally new unknown compound? If you know good the studied microbes, one could check what is the knowledge about their produced compounds and use this knowledge for sequel testing...good luck
First, You must to check whether your antimicrobial compound is a protein or other kind of molecule. (A) If the antimicrobial is a protein. The simple way is that you mix your crude antimicrobial sample with a general protease (e.g. protease K), and check by the well method using a susceptible microorganism. If activity is lost, it is probably that your antimicrobial compound is a protein. Second, if you want to check the molecular mass, check inhibitory activity in situ by SDS-PAGE. Third, search protocols that have been used for purifying antimicrobial proteins, depend on the properties of the protein is the method used for purification. I have used FLPC. HPLC etc.
If your antimicrobial compound is not a proteins, a different strategy you must to use.
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