초록
<P><B>Abstract</B></P><P><B>BACKGROUND</B></P><P>To 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 <I>Escherichia coli</I> Rosetta (DE3) and purified via chromatography. Subsequently, they were co‐crystalized with CaHPO<SUB>4</SUB> at 4°C to form dual‐enzyme@CaHPO<SUB>4</SUB> 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.</P><P><B>RESULTS</B></P><P>At optimum pH of 7.0, the activities of AKR and ADH confined in the dual‐enzyme@CaHPO<SUB>4</SUB> 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.</P><P><B>CONCLUSION</B></P><P>The dual‐enzyme@CaHPO<SUB>4</SUB> 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</P>