100858-34-2

61995-20-8

100858-34-2

General procedure for the synthesis of (R)-N-CBZ-3-hydroxypiperidine from 1-N-Cbz-3-piperidone: In 1 mL of potassium phosphate buffer (100 mM, pH 7.0), a reaction mixture was prepared by adding 200 mM substrate, 1 mM NAD+, 5% (v/v) 2-propanol, and 10 mg of crude enzyme READH. For ChKRED20, the reaction conditions were adjusted to 40% (v/v) 2-propanol and 40°C reaction temperature. The reaction mixture was warmed at 50 °C (or 40 °C for ChKRED20). The reaction progress was monitored by TLC. Upon completion of the reaction, the reaction was terminated by extraction with 1 mL of methyl tert-butyl ether. The organic phase was dried and concentrated with anhydrous sodium sulfate. The conversion and enantiomeric excess were determined by chiral HPLC. The products were purified by silica gel column chromatography and structurally characterized by NMR analysis, spinometry and mass spectrometry.