Effective asymmetric bioreduction of ethyl 4-chloro-3-oxobutanoate to ethyl (R)-4-chloro-3-hydroxybutanoate by recombinant E. coli CCZU-A13 in [Bmim]PF6-hydrolyzate media

It was the first report that the concentrated hydrolyzates from the enzymatic hydrolysis of dilute NaOH (3   wt%)-soaking rice straw at 30   °C was used to form [Bmim]PF 6 -hydrolyzate (50:50, v/v) media for bioconverting ethyl 4-chloro-3-oxobutanoate (COBE) into ethyl ( R )-4-chloro-3-hydroxybutanoate [( R )-CHBE] (>99% e.e. ) with recombinant E. coli CCZU-A13. Compared with pure glucose, the hydrolyzates could promote both initial reaction rate and the intracellular NADH content. Furthermore, emulsifier OP-10 (20   mM) was employed to improve the reductase activity. Moreover, Hp-β-cyclodextrin (0.01   mol Hp-β-cyclodextrin/mol COBE) was also added into this bioreaction system for enhancing the biosynthesis of ( R )-CHBE from COBE by E. coli CCZU-A13 whole-cells. The yield of ( R )-CHBE (>99% e.e. ) from 800   mM COBE was obtained at 100% in the [Bmim]PF 6 -hydrolyzate (50:50, v/v) media by supplementation of OP-10 (20   mM) and Hp-β-CD (8   mM). In conclusion, an effective strategy for the biosynthesis of ( R )-CHBE was successfully demonstrated.