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Applied and Environmental Microbiology, June 2003, p. 3421-3426, Vol. 69, No. 6
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.6.3421-3426.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Metabolic Engineering of Escherichia coli for Production of Enantiomerically Pure (R)-(-)-Hydroxycarboxylic Acids

Sang Yup Lee1,2,3* and Young Lee1,4

Metabolic and Biomolecular Engineering National Research Laboratory, Department of Chemical and Biomolecular Engineering, BioProcess Engineering Research Center,1 Center for Ultramicrochemical Process Systems,2 Department of BioSystems, Korea Advanced Institute of Science and Technology, Yuseong-gu, Daejeon 305-701,3 ChiroBio Inc., Yuseong-gu, Daejeon 305-338, Korea4

Received 24 January 2003/ Accepted 13 March 2003

A heterologous metabolism of polyhydroxyalkanoate (PHA) biosynthesis and degradation was established in Escherichia coli by introducing the Ralstonia eutropha PHA biosynthesis operon along with the R. eutropha intracellular PHA depolymerase gene. By with this metabolically engineered E. coli, enantiomerically pure (R)-3-hydroxybutyric acid (R3HB) could be efficiently produced from glucose. By employing a two-plasmid system, developed as the PHA biosynthesis operon on a medium-copy-number plasmid and the PHA depolymerase gene on a high-copy-number plasmid, R3HB could be produced with a yield of 49.5% (85.6% of the maximum theoretical yield) from glucose. By integration of the PHA biosynthesis genes into the chromosome of E. coli and by introducing a plasmid containing the PHA depolymerase gene, R3HB could be produced without plasmid instability in the absence of antibiotics. This strategy can be used for the production of various enantiomerically pure (R)-hydroxycarboxylic acids from renewable resources.

* Corresponding author. Mailing address: Department of Chemical and Biomolecular Engineering, Korea Advanced Institute of Science and Technology, 373-1 Guseong-dong, Yuseong-gu, Daejeon 305-701, Korea. Phone: 82 42 869 3930. Fax: 82 42 869 3910. E-mail: leesy{at}mail.kaist.ac.kr.

Applied and Environmental Microbiology, June 2003, p. 3421-3426, Vol. 69, No. 6
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.6.3421-3426.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Tseng, H.-C., Martin, C. H., Nielsen, D. R., Prather, K. L. J. (2009). Metabolic Engineering of Escherichia coli for Enhanced Production of (R)- and (S)-3-Hydroxybutyrate. Appl. Environ. Microbiol. 75: 3137-3145 [Abstract] [Full Text]  
  • Uchino, K., Saito, T., Jendrossek, D. (2008). Poly(3-Hydroxybutyrate) (PHB) Depolymerase PhaZa1 Is Involved in Mobilization of Accumulated PHB in Ralstonia eutropha H16. Appl. Environ. Microbiol. 74: 1058-1063 [Abstract] [Full Text]  
  • Gebauer, B., Jendrossek, D. (2006). Assay of Poly(3-Hydroxybutyrate) Depolymerase Activity and Product Determination. Appl. Environ. Microbiol. 72: 6094-6100 [Abstract] [Full Text]  


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