Efficient synthesis of the key chiral alcohol intermediate of Crizotinib using dual‐enzyme@CaHPO4 hybrid nanoflowers assembled by mimetic biomineralization

Abstract

AbstractBACKGROUNDTo develop an environment‐friendly approach for the synthesis of (S)‐1‐(2,6‐dichloro‐3‐fluorophenyl) ethyl alcohol, an intermediate of anti‐cancer drug Crizotinib, aldehyde ketone reductase and alcohol dehydrogenase were overexpressed in Escherichia coli Rosetta (DE3) and purified via chromatography. Subsequently, they were co‐crystalized with CaHPO4 at 4°C to form dual‐enzyme@CaHPO4 hybrid nanoflowers (hNFs) which was then used to catalyze the synthesis of (S)‐1‐(2,6‐dichloro‐3‐fluorophenyl) ethyl alcohol along with the evaluation of its thermal stability and recycling stability.RESULTSAt optimum pH of 7.0, the activities of AKR and ADH confined in the dual‐enzyme@CaHPO4 hybrid nanoflowers were 3.3‐ and 2.1‐fold that of the corresponding free one. The thermos‐stability of confined enzymes was also significantly improved: both enzymes within the hNFs remained more than 80% of initial activities after incubation at 60°C for 8 h, while free enzymes only retained 20% of initial activities under the same treatment conditions. Moreover, AKR and ADH immobilized with a mole ratio of 3:1 confined in hybrid nanoflowers exhibited the highest catalytic activity for the synthesis of chiral ethyl alcohol with a yield up to 90.8% after 12 h. Besides, the final product (S)‐1‐(2,6‐dichloro‐3‐fluorophenyl) ethyl alcohol showed a high ee value of 99.99%. Further, the hybrid nanoflowers retained their initial activity after 16 recycling cycles of synthesis reaction.CONCLUSIONThe dual‐enzyme@CaHPO4 hybrid nanoflowers efficiently catalyzed synthesis of the chiral compound (S)‐1‐(2,6‐dichloro‐3‐fluorophenyl) ethyl alcohol. The method can also be applied to other multi‐enzyme systems and facilitate their cascade reactions and substrate channeling. © 2018 Society of Chemical Industry