extract metabolites from actinomycetes
usually, culture broth was harvested by centrifugation at 4 000 r/min for 15 min. The supernatant was collected and added equal volumes of solvent.
does it matter if we add solvent (methanol) first to culture broth before centrifugation?
We do metabolomics here and the standard extraction we use is 85% MeOH that is usually added before centrifugation. I think it is better to add methanol before centrifugation. This will facilitate the precipitation of proteins that might interfere with metabolomics studies. Also, methanol will cause cell lysis and thus more release of metabolites.
It will also increase the difference between the density of your culture broth (basically by mixing water and ethanol => lower density that water) and your precipitate (celle debris, protein aggregates etc.). Adding methanol will thus ensure a better separation by centrifugation.
But it really depends on what your measurement is based and if methanol can interfere or will enhanced the release of your metabolite of interest as specified by Orwa.
We normally used filter cultured assay to grow bacteria and extract by 40:40:20 (ACN:MeOH:H2O), i will suggest you to use fastprep instead of centrifuge, for metabolites extraction.
I have a metabolomics website (www.metabolomicsonline.com) which collect some resource of microbial metabolomics, you would check it out if interested. Best,
Zhe
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