Enantioselective hydrolysis of ketoprofen amide by Rhodococcus sp. C3II and Rhodococcus erythropolis MP 50

Abstract

The resolution of racemic ketoprofen amide by whole cells of Rhodococcus erythropolis MP 50 and Rhodococcus sp. C3II was studied. With both strains racemic ketoprofen amide was converted to S-ketoprofen with an enantiomeric excess > 97 % at a conversion rate up to 40 % of the theoretical value. The specific activity of strain MP 50 for ketoprofen amide was about 0.12 gmol min -1 and mg of dry weight and the substrate was converted for several hours at a constant rate.

Purification and properties of an amidase from Rhodococcus erythropolis MP50 which enantioselectively hydrolyzes 2-arylpropionamides

“…It was shown earlier that the naproxen amide-hydrolyzing activity in R. erythropolis MP50 was induced after growth of the strain with naproxen nitrile or naproxen amide ( Fig. 1 shows structural formulas) as the sole source of nitrogen (23). To establish the optimal conditions for production of the amidase, strain MP50 was grown in a mineral medium with succinate (10 mM) as the sole source of carbon and energy, and urea, acetamide, isobu- a The usual flow rate was 1 ml/min.…”

“…The isolation of strain MP50 has been described before. The strain was initially classified as Rhodococcus equi (23). A conscientious analysis at the Deutsche Sammlung von Mikroorganismen (DSM) (Braunschweig, Germany) showed that strain MP50 belongs to the species R. erythropolis.…”

“…The growth of bacterial cultures was monitored spectrophotometrically by measuring the optical density at 546 nm (OD 546 ) with a Kontron Uvikon 820 spectrophotometer (Kontron, Eching, Germany). An OD 546 of 1 corresponded to 224 mg weight (dry) of cells per liter of culture (23). The protein content of whole cells was determined by a modification of the method of Schmidt et al (32).…”

“…Aminoethylbenzenesulfonylfluoride was purchased from Boehringer (Mannheim, Germany). The sources of all other chemicals have been described before (5,23,24).…”

“…These cultures were obtained from enrichments with naproxen nitrile as the sole nitrogen source. The isolate Rhodococcus erythropolis MP50 was shown to produce an enantioselective amidase which converted (R,S)-naproxen amide and (R,S)-ketoprofen amide to the corresponding S-acids with enantiomeric excesses above 99% (23,2). In this article, the purification and characterization of this enantioselective amidase are described.…”