I am trying to isolate anti-candida molecules from various isolates, probably actinomycetes. The culture supernatant shows good inhibition zone in well diffusion studies however, when I tried to purify it using precipitation methods by ammonium sulfate I am unable to get activity in precipitate as well as in supernatant. Can anyone suggest how I should modify the method?
You should directly try to treat the fungi by extracts/secreted molecules from actinomyces, or purify the same bimolecule by affinity chromatography or on a HPLC reverse phase column. If you get inhibitory effects largely that any other antibiotic you will get a new compound which may be anti-candia.
Dear Bipinraj,
The method which you are adopting is for proteins. It may be possible that you have non-protein molecules which exhibit anticandida activity. Try to extract them from your supernatant using organic solvents like hexane, ethylacetate etc. I am sure your active molecules will come in either hexane or ethyl acetete. If it will not come and retained in the aqueous layer, then you need to lyophillize the aqueous layer and check its activity further.
- Badges
- Science topic
- Similar topics
- Biological Science
- Biochemistry
- Biochemical Techniques
- Analytical Chemistry
- pH