Efficient bioreductive production of (S)-N-Boc-3-hydroxypiperidine using ketoreductase ChKRED03

Xu, Guangpeng; Wang, Haibo; Wu, Zhong‐Liu

doi:10.1016/j.procbio.2016.04.008

Cited by 23 publications

(10 citation statements)

References 28 publications

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“…The former only achieves a maximum yield of 50%, making the process economically unviable, while the latter appears to be limited because of the lengthy procedure, rather poor yields of the products, and the use of potentially hazardous reagents [1,5,6]. Alternatively, the carbonyl-reductase-catalyzed asymmetric reduction of N -Boc-3-piperidone (NBPO) has gained increasing focus due to its mild reaction conditions, high yield, and remarkable enantioselectivity [4,7,8,9].…”

Section: Introductionmentioning

confidence: 99%

Characterization of a Carbonyl Reductase from Rhodococcus erythropolis WZ010 and Its Variant Y54F for Asymmetric Synthesis of (S)-N-Boc-3-Hydroxypiperidine

et al. 2018

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The recombinant carbonyl reductase from Rhodococcus erythropolis WZ010 (ReCR) demonstrated strict (S)-stereoselectivity and catalyzed the irreversible reduction of N-Boc-3-piperidone (NBPO) to (S)-N-Boc-3-hydroxypiperidine [(S)-NBHP], a key chiral intermediate in the synthesis of ibrutinib. The NAD(H)-specific enzyme was active within broad ranges of pH and temperature and had remarkable activity in the presence of higher concentration of organic solvents. The amino acid residue at position 54 was critical for the activity and the substitution of Tyr54 to Phe significantly enhanced the catalytic efficiency of ReCR. The kcat/Km values of ReCR Y54F for NBPO, (R/S)-2-octanol, and 2-propanol were 49.17 s−1 mM−1, 56.56 s−1 mM−1, and 20.69 s−1 mM−1, respectively. In addition, the (S)-NBHP yield was as high as 95.92% when whole cells of E. coli overexpressing ReCR variant Y54F catalyzed the asymmetric reduction of 1.5 M NBPO for 12 h in the aqueous/(R/S)-2-octanol biphasic system, demonstrating the great potential of ReCR variant Y54F for practical applications.

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Section: Introductionmentioning

confidence: 99%

Characterization of a Carbonyl Reductase from Rhodococcus erythropolis WZ010 and Its Variant Y54F for Asymmetric Synthesis of (S)-N-Boc-3-Hydroxypiperidine

et al. 2018

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“…The absolute configuration of the alcohol was determined by comparing the elution sequence on chiral HPLC with reported data from the literature. 4,27 Preparation of biocatalysts Single colonies of E. coli BL21 (DE3) transformed with the pET28 (+) vectors encoding each of the KREDs (GenBank accession numbers, ChKRED03: KC342003, ChKRED14: KC342014, ChKRED20: KC342020, LKADH: AY267012.1, and LBADH: AJ544275.1) 30,31 were grown overnight at 37 1C in Luria-Bertani (LB) medium containing 50 mg mL À1 kanamycin. The resulting culture (1 mL) was transferred into 200 mL terrific broth (TB) medium containing 50 mg mL À1 kanamycin, and incubated at 37 1C, 180 rpm.…”

Section: Photoenzyme Coupled Asymmetric Reduction Of Prochiral Ketonesmentioning

confidence: 99%

Preparation of chiral aryl alcohols: a controllable enzymatic strategy via light-driven NAD(P)H regeneration

Xing

Liu

et al. 2022

New J. Chem.

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“…At present, it has been reported that several ketoreductases from different sources can be used for the synthesis of (S)-NBHP. For example, Xu et al accomplished the efficient biosynthesis of (S)-NBHP by using cell extract expressing recombinant ketoreductase (ChKRED03) and commercial glucose dehydrogenase pure enzyme [14]. Chen et al reported the biosynthesis of (S)-NBHP, in which the catalysts used were wet cells expressing ketoreductase (YDR541C) and glucose dehydrogenase alone [15].…”

Section: Introductionmentioning

confidence: 99%

Efficient Asymmetric Synthesis of (S)-N-Boc-3-hydroxypiperidine by Coexpressing Ketoreductase and Glucose Dehydrogenase

et al. 2022

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(S)-N-Boc-3-hydroxypiperidine is an important intermediate of the anticancer drug ibrutinib and is mainly synthesized by the asymmetric reduction catalyzed by ketoreductase coupled with glucose dehydrogenase at present. In this study, the coexpression recombinant strains E. coli/pET28-K-rbs-G with single promoter and E. coli/pETDuet-K-G with double promoters were first constructed for the coexpression of ketoreductase and glucose dehydrogenase in the same cell. Then, the catalytic efficiency of E. coli/pET28-K-rbs-G for synthesizing (S)-N-Boc-3-hydroxypiperidine was found to be higher than that of E. coli/pETDuet-K-G due to the more balanced activity ratio and higher catalytic activity. On this basis, the catalytic conditions of E. coli/pET28-K-rbs-G were further optimized, and finally both the conversion of the reaction and the optical purity of the product were higher than 99%. In the end, the cell-free extract was proved to be a better catalyst than the whole cell with the improved catalytic efficiency of different recombinant strains. This study developed a better coexpression strategy for ketoreductase and glucose dehydrogenase by investigating the effect of activity ratios and forms of the biocatalysts on the catalytic efficiency deeply, which provided a research basis for the efficient synthesis of chiral compounds.

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Efficient bioreductive production of (S)-N-Boc-3-hydroxypiperidine using ketoreductase ChKRED03

Cited by 23 publications

References 28 publications

Characterization of a Carbonyl Reductase from Rhodococcus erythropolis WZ010 and Its Variant Y54F for Asymmetric Synthesis of (S)-N-Boc-3-Hydroxypiperidine

Characterization of a Carbonyl Reductase from Rhodococcus erythropolis WZ010 and Its Variant Y54F for Asymmetric Synthesis of (S)-N-Boc-3-Hydroxypiperidine

Preparation of chiral aryl alcohols: a controllable enzymatic strategy via light-driven NAD(P)H regeneration

Efficient Asymmetric Synthesis of (S)-N-Boc-3-hydroxypiperidine by Coexpressing Ketoreductase and Glucose Dehydrogenase

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