Abstract:(S)-N-Boc-3-hydroxypiperidine ((S)-NBHP) is a key chiral intermediate for the synthesis of ibrutinib treating mantle cell lymphoma. Asymmetric reduction of N-Boc-3-piperidone (NBPO) catalyzed by alcohol dehydrogenase is the most promising method for (S)-NBHP synthesis. After screening of the alcohol dehydrogenase library, KpADH from Kluyveromyces polyspora showed excellent catalytic efficiency on NBPO, with specific activity of pure enzyme as high as 83.9 U/mg and the product enantiomeric excess value of 97.0% (S). Coupled cofactor regeneration reaction was established by introducing glucose dehydrogenase, and the effects of monoaquous phase, biphasic phase, ionic liquids and different substrate concentrations on the conversion were explored. The results indicated that high concentration product (S)-NBHP had inhibitory effect on KpADH, and the inhibitory kinetic analysis showed that it belonged to non-competitive inhibition. The inhibition of (S)-NBHP on KpADH catalytic reaction could be reduced and the conversion efficiency could be improved by using single monoaquous phase catalysis. The inhibition of asymmetric reduction of NBPO catalyzed by alcohol dehydrogenase was studied, which could provide guidance for the enzymatic preparation of (S)-NBHP.
