Hello Haseena,

An activity in the cell free supernatant could be because of any extracellular secretion of the organism. Hence you need to go further with the purification of the crude supernatant for determining the agent responsible. Bioactive organic lead can be determined through liquid-liquid solvent extraction (polar or non-polar) and for enzyme or bioactive peptides proceed with protein purification. Thereafter, you can confirm the bioactivity in these partially purified fractions.

It's very hard to answer.

If you don't have any suspicious structure, you have a long way.

I suggest you give more information in otherwise you have to remove agent step by step and run many tests to figure out what component you face up and then run tests for figure out right structures.

Regards.

A bio-active compound could either be a proteinacious, carbohydrate or lipidic substance. I am not aware of any enzyme that contains lipids. So, if the supernatant tests positive for the biochemical test for lipids, say ethanol emulsion or any other, then you can rule out enzymes as being the antibacterial compounds and concentrate on other bio-active compounds.

Thanks