Efficient synthesis of (S)-N-Boc-3-hydroxypiperidine using an (R)-specific carbonyl reductase from Candida parapsilosis

World J Microbiol Biotechnol. 2017 Mar;33(3):61. doi: 10.1007/s11274-016-2189-y. Epub 2017 Feb 27.


(S)-N-Boc-3-hydroxypiperidine (S-NBHP) is a critical chiral intermediate in the synthesis of pharmaceuticals, including ibrutinib, the active pharmaceutical ingredient of the new drug Imbruvica approved for the treatment of lymphoma. An (R)-specific carbonyl reductase from Candida parapsilosis (CprCR, also known as R-specific alcohol dehydrogenase) that catalyzes asymmetric reduction to produce (S)-N-Boc-3-hydroxypiperidine (S-NBHP) was identified for the first time. When co-expressed with a glucose dehydrogenase from Bacillus megaterium in Escherichia coli Rosetta (DE3), recombinant crude enzyme exhibited an activity of 9 U/mg with N-Boc-3-piperidone as the substrate and 12 U/mg with glucose as the substrate. The biocatalysis of N-Boc-3-piperidone to S-NBHP using recombinant whole-cell biocatalysts was processed in a water/butyl acetate system as well as an aqueous monophasic system without extra NAD+/NADH. This process showed great commercial potential, with a 100 g/l substrate concentration and a whole cells loading (w/v) of 10%, with the conversion of 97.8% and an e.e. of 99.8% in an aqueous monophasic system.

Keywords: (S)-N-Boc-3-hydroxypiperidine; Carbonyl reductase; Commercial potential; Recombinant whole-cell.

MeSH terms

  • Alcohol Dehydrogenase / genetics*
  • Alcohol Dehydrogenase / metabolism*
  • Bacillus megaterium / enzymology
  • Bacillus megaterium / genetics
  • Biocatalysis
  • Candida / enzymology*
  • Candida / genetics
  • Cloning, Molecular
  • Escherichia coli / genetics
  • Fungal Proteins / genetics
  • Fungal Proteins / metabolism
  • Glucose 1-Dehydrogenase / genetics
  • Glucose 1-Dehydrogenase / metabolism
  • Piperidines / metabolism*
  • Recombinant Proteins / metabolism


  • Fungal Proteins
  • Piperidines
  • Recombinant Proteins
  • Alcohol Dehydrogenase
  • Glucose 1-Dehydrogenase