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Applied and Environmental Microbiology, February 2005, p. 713-720, Vol. 71, No. 2
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.2.713-720.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Kinetic Studies and Biochemical Pathway Analysis of Anaerobic Poly-(R)-3-Hydroxybutyric Acid Synthesis in Escherichia coli

Ross Carlson,{dagger} Aaron Wlaschin, and Friedrich Srienc*

Department of Chemical Engineering and Materials Science and BioTechnology Institute, University of Minnesota, St. Paul, Minnesota

Received 19 August 2004/ Accepted 19 September 2004

Poly-(R)-3-hydroxybutyric acid (PHB) was synthesized anaerobically in recombinant Escherichia coli. The host anaerobically accumulated PHB to more than 50% of its cell dry weight during cultivation in either growth or nongrowth medium. The maximum specific PHB production rate during growth-associated synthesis was approximately 2.3 ± 0.2 mmol of PHB/g of residual cell dry weight/h. The by-product secretion profiles differed significantly between the PHB-synthesizing strain and the control strain. PHB production decreased acetate accumulation for both growth and nongrowth-associated PHB synthesis. For instance under nongrowth cultivation, the PHB-synthesizing culture produced approximately 66% less acetate on a glucose yield basis as compared to a control culture. A theoretical biochemical network model was used to provide a rational basis to interpret the experimental results like the fermentation product secretion profiles and to study E. coli network capabilities under anaerobic conditions. For example, the maximum theoretical carbon yield for anaerobic PHB synthesis in E. coli is 0.8. The presented study is expected to be generally useful for analyzing, interpreting, and engineering cellular metabolisms.

* Corresponding author. Mailing address: Department of Chemical Engineering and Materials Science and BioTechnology Institute, University of Minnesota, 240 Gortner Laboratory, 1479 Gortner Ave., St. Paul, MN 55108. Phone: (612) 624-9776. Fax: (612) 625-1700. E-mail: srienc{at}umn.edu.

{dagger} Present address: Center for Biofilm Engineering, Montana State University, Bozeman, MT 59717.

Applied and Environmental Microbiology, February 2005, p. 713-720, Vol. 71, No. 2
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.2.713-720.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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  • Nikel, P. I., Pettinari, M. J., Galvagno, M. A., Mendez, B. S. (2006). Poly(3-Hydroxybutyrate) Synthesis by Recombinant Escherichia coli arcA Mutants in Microaerobiosis. Appl. Environ. Microbiol. 72: 2614-2620 [Abstract] [Full Text]  


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