Hi, i'm workint now on Actinobacteria (Streptomyces) cultured on ISP2, and i need to extract it's metabolites to reveal its insecticidal effects, can anyone guide me which protocolor or procedure to follow? and which is the best medium be used to increase metabolites production??
normally we separate the culture broth in biomass (mycelium) and culture filtrate by multiple sheet filtration. The mycelium was extracted 2-4 times with methanol-acetone 1:1, the culture filtrate was extracted 3 to 5 times with ethylacetate (pH 4-5) or we perform XAD 16 chromatography (2 l column for 20 l culture filtrate), followed by ethylacete extraction of the aqueous residue after concentration of the eluate.
How much product you can isolate at last depends on the production of the compound. We have isolated amounts of 5-200 mg from a 20 l batch fermentation. But it could be more or less...
You can read more in the attached paper in the section "fermentation and isolation".
DOI: 10.1038/ja.2011.53
Hope this helps
Thanks Wiwiek Harsonowati, i appreciate your response and i will look at the paper.
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