Isolation of Bradyoxetin

Preparation of CDF

1.                  Bacteria cultures were grown up in Minimal medium to
late log phase (OD600 = 2.0).

2.                  Equal volume of ethyl acetate (Optima Grade, Fisher
Scientific) was then added and incubated, with shaking at 200 rpm, for 1

3.                  The flasks were then allowed to sit for 1 h, or as
long as it takes to separate the aqueous phase from the ethyl acetate
phase.  Should the layers fail to separate, additional ethyl acetate is
added and step 3 is repeated.

4.                  Remove the top layer containing the ethyl acetate
using a glass pipette, and store in glass container.  The ethyl acetate
is removed by blow-drying, using the house air-line.

5.                  The concentrated extract, containing CDF, is then
stored at -20 C.

HPLC analyses of CDF