Biotechnology Progress, Vol.33, No.3, 612-620, 2017
Large-Scale Synthesis of Tert-Butyl (3R,5S)-6-Chloro-3,5-Dihydroxyhexanoate by a Stereoselective Carbonyl Reductase with High Substrate Concentration and Product Yield
To biosynthesize the (3R, 5S)-CDHH in an industrial scale, a newly synthesized stereoselective short chain carbonyl reductase (SCR) was successfully cloned and expressed in Escherichia coli. The fermentation of recombinant E. coli harboring SCR was carried out in 500 L and 5000 L fermenters, with biomass and specific activity of 9.7 g DCW/L, 15749.95 U/g DCW, and 10.97 g DCW/L, 19210.12 U/g DCW, respectively. The recombinant SCR was successfully applied for efficient production of (3R,5S)-CDHH. The scale-up synthesis of (3R,5S)-CDHH was performed in 5000 L bioreactor with 400 g/L of (S)-CHOH at 30 degrees C, resulting in a space-time yield of 13.7 mM/h/g DCW, which was the highest ever reported. After isolation and purification, the yield and d.e. of (3R,5S)-CDHH reached 97.5% and 99.5%, respectively. (C) 2017 American Institute of Chemical Engineers.
Keywords:carbonyl reductase;tert-butyl (S)-6-chloro-5-hydroxy-3-oxohexanoate;tert-butyl (3R,5S)-6-chloro-3,5-dihydroxyhexanoate;asymmetric reduction;space-time yield