Applied Microbiology and Biotechnology

, Volume 79, Issue 4, pp 633–641

Biosynthesis of enantiopure (S)-3-hydroxybutyric acid in metabolically engineered Escherichia coli

Applied Genetics and Molecular Biotechnology

DOI: 10.1007/s00253-008-1473-7

Cite this article as:
Lee, SH., Park, S.J., Lee, S.Y. et al. Appl Microbiol Biotechnol (2008) 79: 633. doi:10.1007/s00253-008-1473-7

Abstract

A biosynthetic pathway for the production of (S)-3-hydroxybutyric acid (S3HB) from glucose was established in recombinant Escherichia coli by introducing the β-ketothiolase gene from Ralstonia eutropha H16, the (S)-3-hydroxybutyryl-CoA dehydrogenase gene from R. eutropha H16, or Clostridium acetobutylicum ATCC824, and the 3-hydroxyisobutyryl-CoA hydrolase gene from Bacillus cereus ATCC14579. Artificial operon consisting of these genes was constructed and was expressed in E. coli BL21 (DE3) codon plus under T7 promoter by isopropyl β-d-thiogalactoside (IPTG) induction. Recombinant E. coli BL21 (DE3) codon plus expressing the β-ketothiolase gene, the (S)-3-hydroxybutyryl-CoA dehydrogenase gene, and the 3-hydroxyisobutyryl-CoA hydrolase gene could synthesize enantiomerically pure S3HB to the concentration of 0.61 g l−1 from 20 g l−1 of glucose in Luria–Bertani medium. Fed-batch cultures of recombinant E. coli BL21 (DE3) codon plus were carried out to achieve higher titer of S3HB with varying induction time and glucose concentration during fermentation. Protein expression was induced by addition of 1 mM IPTG when cell concentration reached 10 and 20 g l−1 (OD600 = 30 and 60), respectively. When protein expression was induced at 60 of OD600 and glucose was fed to the concentration of 15 g l−1, 10.3 g l−1 of S3HB was obtained in 38 h with the S3HB productivity of 0.21 g l−1h−1. Lowering glucose concentration to 5 g l−1 and induction of protein expression at 30 of OD600 significantly reduced final S3HB concentration to 3.7 g l−1, which also resulted in the decrease of the S3HB productivity to 0.05 g l−1h−1.

Keywords

(S)-3-hydroxybutyric acid Escherichia coli Metabolic engineering Fed-batch culture 

Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  1. 1.Corporate R&DLG Chem, Ltd./Research ParkDaejeonRepublic of Korea
  2. 2.Metabolic and Biomolecular Engineering National Research Laboratory, Department of Chemical and Biomolecular Engineering (BK21 Program), BioProcess Engineering Research Center, Center for Systems and Synthetic BiotechnologyInstitute for the BioCenturyDaejeonRepublic of Korea
  3. 3.Department of Bio and Brain Engineering, and Bioinformatics Research CenterKorea Advanced Institute of Science and TechnologyDaejeonRepublic of Korea
  4. 4.School of Chemical Engineering & BioengineeringUniversity of UlsanUlsanRepublic of Korea

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