(S)-1-Cbz-3-pyrrolidinol 95

130312-02-6

100858-32-0

General procedure for the synthesis of benzyl (S)-3-hydroxypyrrolidine-1-carboxylate using N-Cbz-3-pyrrolidone as starting material: a reaction mixture containing 200 mM substrate, 1 mM NAD+, 5% (v/v) 2-propanol, and 10 mg of the crude enzyme READH was prepared in 1 mL of potassium phosphate buffer (100 mM, pH 7.0) and warmed at 50 °C. For ChKRED20, 40% (v/v) 2-propanol was used and the reaction was carried out at 40°C. The reaction progress was monitored by TLC and the reaction was terminated by extraction with methyl tert-butyl ether (1 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated. Samples were analyzed by chiral HPLC to determine conversion and enantiomeric excess. The final product was purified by silica gel column chromatography and the structure was confirmed by NMR, spin determination and mass spectrometry.