Efficient synthesis of (S)-N-Boc-3-hydroxypiperidine using an (R)-specific carbonyl reductase from Candida parapsilosis
- 166 Downloads
(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.
KeywordsCarbonyl reductase Commercial potential Recombinant whole-cell (S)-N-Boc-3-hydroxypiperidine
This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.
- Makrides SC (1996) Strategies for achieving high-level expression of genes in Escherichia coli. Microbiol Rev 60:512–538Google Scholar
- Musa MM, Phillips RS (2011) Recent advances in alcohol dehydrogenase-catalyzed asymmetric production of hydrophobic alcohols. Catalscitechnol 1:1311–1323Google Scholar
- Shen W, Shen Z, Hu Z, Lin Y et al (2013) Synthesis of (S)-1-Boc-3-hydroxypiperidine. Chin J Pharm 44:436–438Google Scholar
- Zhu W, Wang B, Wu H, Li B (2015) A Chemo-enzyme method to synthesis of (S)-t-butyl 3-hydroxypiperidine-1-carboxylate. Chin J Pharm 46:349–350Google Scholar