Microbiology List: purification of an antifungal compound

SPPalaniappan at nagarjunagroup.com SPPalaniappan at nagarjunagroup.com
Mon Jun 11 01:31:32 EST 2001


----- Forwarded by S P Palaniappan/DIRO/WAR/NARDI/NAGARJUNA on 06/11/2001
12:05 PM -----
                                                                           
                    S P                                                    
                    Palaniappan          To:     microbio at net.bio.net      
                                         cc:     jakku at mrna.tn.nic.in      
                    06/07/2001           Subject:     Re: Microbiology     
                    10:17 AM             List: purification of an          
                                         antifungal compound(Document      
                                         link: S P Palaniappan)            
                                                                           



Dear Mr. Jayakumar,

The culture supernatant should be concentrated using rotoevaporator/freeze
dryer to oily past or dry powder. This concentrated materials can be
extracted with different solvent system. Based on solubility and polarity
the compound(s) can come in to the solvent. The presence of compound(s) can
be identified by TLC. Based on the RF value, the compound(s) will be
purified by column chromatography using silica gel. The column will be
eluted with different solvents, based on TLC experience. Further TLC should
be carried out for each fraction of eluate to determine the presence of
compound. Fractions showing single spot in TLC can be pooled and
concentrated using rotoevaporator and bioassayed with fungal pathogens.
Through this downstream method, compound can be purified effectively.

For selection of solvent system and TLC, you may consult with any organic
chemist, who is working on natural product.

Once the compound is purified further characterization can be carried out
using UV, IR, NMR, Mass spectral analyses and X-ray crystallography can be
used to elucidate the chemical structure of the compound.

Dr. N. Mathivanan
Program Leader - Microbial Metabolites
Nagarjuna Agricultural Research and Development Institute
61, Nagarjuna Hills
Panjagutta
Hyderabad - 500 082
E-mail: nardi at nagarjunagroup.com
prabhamathi at hotmail.com



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