earticle

영어

Ketoprofen [(R, S)-2-(3-benzoylphenyl) propionic acid, C16H14O3] is one of the non-steroidal anti-inflammatory drug (NSAID) with analgesic, antipyretic, and anti-inflammatory effects, which has been known to inhibit the production of prostaglandin. Specifically, (S)-ketoprofen is a main compound for these effects, while the (R)-ketoprofen has unnecessary side effects. Therefore, several methods to prepare (S)-ketoprofen are investigated including asymmetric synthesis, enzymatic, and chemical resolution with different effectiveness. In our study we have screened a strain Bacillus gelatini which has a positive activity to a certain extent could asymmetricly hydrolyzes the rac-ketoprofen chloroethyl ester to (s)-ketoprofen(ee%≈ 75%). So our purpose of the study is to clone and express this specific esterase gene. As a result, at first we have cloned the esterase gene by PCR, and then ligation with pBluescriptⅡ SK+, transformation with E.coli XL1-blue and expressed various proteins in transformants. The transformants have been tested the esterase activity by esterase activity staining and analysised the hydrolyzation of rac-ketoprofen to (s)-ketoprofen by HPLC.