I have been growing my actino at 28 degrees on a 200rpm shaker for 12 days and the supernatant is not showing any activity against the test bacterium (M. aurum). I have previously done overlay assays i.e. streaked out the actino and overlayed with sloppy M.aurum and have observed activity. Is it likely that my actino is not producing in liquid culture?
You could precipitate the secondary metabolite after separating the cellular mat ,, by
treatment with 1:1 ratio solvents ,,i.e/ chloroform: Ethanol /methanol.
Then the precipitate could be separated and then dissolved and concentrated in minicon cells .
the concentrated and partially purified product could be tested for its anti-microbial
activity by dissolving the paste and soaking the mini-filter disks with known volume and placing them over the indicator bacterial lawn and incubation to assay activity.
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DR. MAY AL-DOORI
Secondary metabolite production is not triggered in all fermentation media. It could be usefull to carry out fermentation of "your" actino in diverse media containing different carbon and nitrogen sources. Prepare a seed colture to inoculate all the media. After initial growth test every day for AB activity (either on supernatant and mycelia extracted with methanol or acetonitrile) . In the mean time check again activity of the solid colture .
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