title: Efficient Preparation of (S)-N-Boc-3-Hydroxylpiperidine Through Bioreduction by a Thermostable Aldo-KetoReductase.
abstract: (S)-N-Boc-3-hydroxypiperidine ((S)-NBHP) is a key pharmaceutical intermediate and the chiral source in synthesizing Imbruvica, which is a newly approved drug in lymphoma therapy by targeting Bruton's tyrosine kinase (BTK). Current chemical synthesis of (S)-NBHP suffered from the need of noble metal catalyst and low yield. The single reported bioconversion of (S)-NBHP was achieved by using recombinant ketoreductase, but enzyme sequence was kept confidential and the catalytic process suffered from the thermodeactivation and substrate inhibition. In the current study, we presented a thermostable aldo-keto reductase (AKR)-AKR-43-which showed high activity toward N-Boc-3-piperidone (NBP) to produce (S)-NBHP, high enantioselectivity, and no substrate inhibition. The molecular simulations demonstrated the structural rationale for the enantioselectivity of AKR-43 toward NBP and supported the classic ordered two-step catalytic mechanism. The catalytic process was achieved by using glucose dehydrogenase (GDH) for cofactor recycling, and the optimal reaction conditions were determined to be 30 °C and pH 7.5. Within a reaction time of 16 h, the 16 % substrate concentration (w/w), over 99 % ee and under 3.5 % of enzyme loading (w/w) characterized a high efficiency process with promising industrial values.
authors.affiliation: Lab of Biocatalysis, Hangzhou Normal University, Hangzhou, Zhejiang, 311121, China.
authors.forename: Mengyan
authors.initials: M
authors.lastname: He
authors.affiliation: Lab of Biocatalysis, Hangzhou Normal University, Hangzhou, Zhejiang, 311121, China.
authors.forename: Shuo
authors.initials: S
authors.lastname: Zhou
authors.affiliation: Lab of Biocatalysis, Hangzhou Normal University, Hangzhou, Zhejiang, 311121, China.
authors.forename: Maolin
authors.initials: M
authors.lastname: Cui
authors.affiliation: Apeloa Pharmaceutical Co., Ltd., Dongyang, Zhejiang, 322118, China.
authors.forename: Xiaolu
authors.initials: X
authors.lastname: Jin
authors.affiliation: Lab of Biocatalysis, Hangzhou Normal University, Hangzhou, Zhejiang, 311121, China.
authors.forename: Dunyue
authors.initials: D
authors.lastname: Lai
authors.affiliation: Lab of Biocatalysis, Hangzhou Normal University, Hangzhou, Zhejiang, 311121, China.
authors.forename: Shuangling
authors.initials: S
authors.lastname: Zhang
authors.affiliation: Institute of Aging Research, School of Medicine, Hangzhou Normal University, Hangzhou, Zhejiang, 311121, China. zhgwang@hznu.com.
authors.forename: Zhiguo
authors.initials: Z
authors.lastname: Wang
authors.affiliation: Lab of Biocatalysis, Hangzhou Normal University, Hangzhou, Zhejiang, 311121, China. zchenhznu@126.com.
authors.forename: Zhenming
authors.initials: Z
authors.lastname: Chen
mesh_terms: D000449:Aldehyde Reductase / Q000737:chemistry / Q000378:metabolism*; D000074408:Aldo-Keto Reductases; D001412:Bacillus subtilis / Q000201:enzymology; D055162:Biocatalysis; D004795:Enzyme Stability; D056004:Molecular Dynamics Simulation; D010084:Oxidation-Reduction; D010880:Piperidines / Q000737:chemistry / Q000378:metabolism*; D011487:Protein Conformation; D013237:Stereoisomerism; D013379:Substrate Specificity; D013696:Temperature
chemical_list: C000617633:N-tert-butoxycarbonyl-3-hydroxypiperidine; D010880:Piperidines; D000074408:Aldo-Keto Reductases; D000449:Aldehyde Reductase
keywords: (S)-N-Boc-3-hydroxypiperidine; Aldo-keto reductase; Bioconversion; Molecular simulation; N-Boc-3-piperidone