Applied Microbiology and Biotechnology

, Volume 84, Issue 3, pp 487–497

Comparison of engineered Saccharomyces cerevisiae and engineered Escherichia coli for the production of an optically pure keto alcohol

  • Nádia Skorupa Parachin
  • Magnus Carlquist
  • Marie F. Gorwa-Grauslund
Biotechnological Products and Process Engineering

DOI: 10.1007/s00253-009-1964-1

Cite this article as:
Skorupa Parachin, N., Carlquist, M. & Gorwa-Grauslund, M.F. Appl Microbiol Biotechnol (2009) 84: 487. doi:10.1007/s00253-009-1964-1

Abstract

In this study, the production of enantiomerically pure (1R,4S,6S)-6-hydroxy-bicyclo[2.2.2]octane-2-one ((−)-2) through stereoselective bioreduction was used as a model reaction for the comparison of engineered Saccharomyces cerevisiae and engineered Escherichia coli as biocatalysts. For both microorganisms, over-expression of the gene encoding the NADPH-dependent aldo-keto reductase YPR1 resulted in high purity of the keto alcohol (−)-2 (>99% ee, 97–98% de). E. coli had three times higher initial reduction rate but S. cerevisiae continued the reduction reaction for a longer time period, thus reaching a higher conversion of the substrate (95%). S. cerevisiae was also more robust than E. coli, as demonstrated by higher viability during bioreduction. It was also investigated whether the NADPH regeneration rate was sufficient to supply the over-expressed reductase with NADPH. Five strains of each microorganism with varied carbon flux through the NADPH regenerating pentose phosphate pathway were genetically constructed and compared. S. cerevisiae required an increased NADPH regeneration rate to supply YPR1 with co-enzyme while the native NADPH regeneration rate was sufficient for E. coli.

Keywords

BioreductionAsymmetric carbonyl reductionChiral alcoholRecombinant Saccharomyces cerevisiaeRecombinant Escherichia coliAldo-keto reductaseYPR1(1R,4S,6S)-6-hydroxy-bicyclo[2.2.2]octane-2-one

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Nádia Skorupa Parachin
    • 1
  • Magnus Carlquist
    • 1
  • Marie F. Gorwa-Grauslund
    • 1
  1. 1.Department of Applied Microbiology, Center for Chemistry and Chemical engineeringLund UniversityLundSweden