Dear Manjunath,

Inoculate the fungi (pure) into about  40 Erlenmeyer flasks (1L) containing potato dextrose broth (PDB) media (400 mL) and incubate at room temperature for ten days and incubate while shaking every other day (about 90 rpm) for about 20 days. After four weeks filter the broth and partitioned with EtOAc and evaporate under reduced pressure to obtain the Crude extract 1 and extract the residual mycelium with EtOAc and MeOH using ultra sonicator to give EtOAc and MeOH extracts 2 and 3 respectively. By comparing TLC of these extracts and further purification of crude by CC, PTLC, HPLC etc.  you can easily isolate the compounds. 

The cultural condition in dark with room temperature. 

Thank you Hui Song.

Yes Manjunath, incubate the flasks under dark conditions at room temperature

Dear Manjunath

First of all extract the crude metabolite from fungal broth through EtOAc and dry it through rotatory evaporator and than mix silica gel (60 meash) and load that silia + metabolite in to CC and start the purification from hexane and  isolation of compound when stop separated,  increase polarity by mixing 9:1 (hex: EtoAc) and like wise increase the polarity upto 9:1 (EtOac : Hex) and finally elute the metabolite in 100% ETOAc.

Dear Manjunath, 

Protocol given by Ashish and M.M. Qader is the best way for isolation of metabolites. 

Precaution: Clean your rotatory evaporator with solvent.