Recombinant E. coli BL21 cells with wet weight of 0.5–2.0 g
were resuspended in 10 ml 100mM potassium phosphate buffer
(pH 6.5) containing 20% (v/v) bis-(2-ethylhexyl) phthalate (BEHP)
and 10mg substrate. The reaction was carried out at 30 ◦C for
24 h with gyratory shaking at 220rpm and terminated by extraction
with ether. The combined organic extracts were dried with
anhydrous sodium sulfate, concentrated under reduced pressure
and subjected to high performance liquid chromatography (HPLC)
analysis performed on a Shimadzu Prominence LC-20AD system
connected to a PDA-detector. Chemical yields were analyzed by
reverse-phase HPLC on a Luna C18 column (4.6mm×150mm,Phenomenex,
Torrance, CA, USA). Optical purities were determined by
chiral HPLC using Daicel Chiralcel OD-H (compound 4) and OJ-H
(compound 3), or Chiralpak AD-H (compounds 1 and 2) and AS-H
(compounds 5–11) columns.