Application of comparative proteome analysis to reveal influence of cultivation conditions on asymmetric bioreduction of β-keto ester by Saccharomyces cerevisiae

  • Jinping Lin
  • Qinghai Liu
  • Erzheng Su
  • Dongzhi Wei
  • Shengli Yang
Genomics and Proteomics

DOI: 10.1007/s00253-008-1602-3

Cite this article as:
Lin, J., Liu, Q., Su, E. et al. Appl Microbiol Biotechnol (2008) 80: 831. doi:10.1007/s00253-008-1602-3

Abstract

Industrial bakers’ yeast strain Saccharomyces cerevisiae LH1 was selected for asymmetric reduction of ethyl benzoylacetate to (S)-ethyl 3-hydroxy-3-phenylpropionate. Higher reductive efficiency and higher cofactor availability were obtained with the alternation of cultivation condition (mainly growth medium). Compared to the bioreduction by yeast cells grown in malt extract (ME) medium, the concentration of substrate was increased 25-fold (up to 15.6 g/l) in the yeast peptone dextrose (YPD)-grown cells mediated bioreduction with 97.5% of enantioselective excess of (S)-product. The proteomic responses of S. cerevisiae LH1 cells to growth in aerobic batch cultures fed with either YPD or ME medium were examined and compared. Among the relative quantities of 550 protein spots in each gel, changes were shown in the expression level of 102 intracellular proteins when comparing YPD gel to ME gel. Most of the identified proteins were involved in energy metabolism and several cellular molecular biosynthetic pathway and catabolism. For YPD-grown yeast cells, not only enzymes involved in nicotinamide adenine dinucleotide phosphate regeneration, especially 6-phosphogluconate dehydrogenase, but also alcohol dehydrogenase 1 and d-arabinose 1-dehydrogenase which had been demonstrated activity toward ethyl benzoylacetate to (S)-hydroxy ester were significantly upregulated. These changes provided us insight in the way the yeast cells adapted to a change in cultivation medium and regulated its catalytic efficiency in the bioreduction.

Keywords

Asymmetric bioreductionSaccharomyces cerevisiaeGrowth mediumEthyl benzoylacetateProteome analysis

Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Jinping Lin
    • 1
  • Qinghai Liu
    • 1
  • Erzheng Su
    • 1
  • Dongzhi Wei
    • 1
  • Shengli Yang
    • 1
  1. 1.State Key Laboratory of Bioreactor Engineering, New World Institute of BiotechnologyEast China University of Science and TechnologyShanghaiPeople’s Republic of China