Ampicillin → Methyl (R)-(-)-mandelate

Participants

Reaction Steps

The transformed strain XL10-Gold (pTrcFadLPL) prepared in Example 3 was inoculated again in 250 mL Erlenmeyer flask including 100 mL of LB liquid medium containing 50 μg/L of ampicillin, and then cultured at 37° C. Gene expression was induced by adding 0.1 mM of IPTG, when absorbance was 0.5 at 600 nm wavelength by spectrophotometer. 5 hours after inducing the expression, culture broth was centrifuged at 4° C., 600 rpm for 5 minutes to collect cells, and then, the collected cells were washed with 50 mM Tris-HCl solution (pH 8.0), followed by resuspending in 3 mL of buffer. Said re-suspension was put into a 10 mL flask, to which 30 mg of racemic mandelic acid methyl ester was added and allowed to react for 48 hours with stirring. Then, the suspension was centrifuged at 4° C. and 6000 rpm for 5 minutes to remove cells. The supernatant was collected and adjusted to a pH of 3.0. After extraction of the supernatant with 10 mL of ethyl acetate and hexane, the extract was analyzed by liquid chromatography. The analysis results showed that (S)-mandelic acid and (R)-mandelic acid methyl ester were obtained.

Provenance & Evidence