title: Efficient synthesis of (S)-N-Boc-3-hydroxypiperidine using an (R)-specific carbonyl reductase from Candida parapsilosis.
abstract: (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.
authors.affiliation: College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, No.30 Puzhu Road, Nanjing, 211816, People's Republic of China.
authors.forename: Jingjing
authors.initials: J
authors.lastname: Chen
authors.affiliation: College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, No.30 Puzhu Road, Nanjing, 211816, People's Republic of China. yanming@njtech.edu.cn.
authors.forename: Ming
authors.initials: M
authors.lastname: Yan
authors.affiliation: College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, No.30 Puzhu Road, Nanjing, 211816, People's Republic of China.
authors.forename: Lin
authors.initials: L
authors.lastname: Xu
mesh_terms: D000426:Alcohol Dehydrogenase / Q000235:genetics* / Q000378:metabolism*; D001410:Bacillus megaterium / Q000201:enzymology / Q000235:genetics; D055162:Biocatalysis; D002175:Candida / Q000201:enzymology* / Q000235:genetics; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D042843:Glucose 1-Dehydrogenase / Q000235:genetics / Q000378:metabolism; D010880:Piperidines / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism
chemical_list: D005656:Fungal Proteins; D010880:Piperidines; D011994:Recombinant Proteins; D000426:Alcohol Dehydrogenase; D042843:Glucose 1-Dehydrogenase
keywords: (S)-N-Boc-3-hydroxypiperidine; Carbonyl reductase; Commercial potential; Recombinant whole-cell