This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Klinke, S.
Right arrow Articles by Kessler, B.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Klinke, S.
Right arrow Articles by Kessler, B.
Agricola
Right arrow Articles by Klinke, S.
Right arrow Articles by Kessler, B.

 Previous Article  |  Next Article 

Applied and Environmental Microbiology, February 1999, p. 540-548, Vol. 65, No. 2
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Production of Medium-Chain-Length Poly(3-Hydroxyalkanoates) from Gluconate by Recombinant Escherichia coli

Stefan Klinke, Qun Ren, Bernard Witholt,* and Birgit Kessler

Institute of Biotechnology, Swiss Federal Institute of Technology Zurich, CH-8093 Zurich, Switzerland

Received 8 September 1998/Accepted 23 November 1998

It was shown recently that recombinant Escherichia coli, defective in the beta -oxidation cycle and harboring a medium-chain-length (MCL) poly(3-hydroxyalkanoate) (PHA) polymerase-encoding gene of Pseudomonas, is able to produce MCL PHA from fatty acids but not from sugars or gluconate (S. Langenbach, B. H. A. Rehm, and A. Steinbüchel, FEMS Microbiol. Lett. 150:303-309, 1997; Q. Ren, Ph.D. thesis, ETH Zürich, Zürich, Switzerland, 1997). In this study, we report the formation of MCL PHA from gluconate by recombinant E. coli. By introduction of genes coding for an MCL PHA polymerase and the cytosolic thioesterase I ('thioesterase I) into E. coli JMU193, we were able to engineer a pathway for the synthesis of MCL PHA from gluconate. We used two expression systems, i.e., the bad promoter and alk promoter, for the 'thioesterase I- and PHA polymerase-encoding genes, respectively, which enabled us to modulate their expression independently over a range of inducer concentrations, which resulted in a maximum MCL PHA accumulation of 2.3% of cell dry weight from gluconate. We found that the amount of PHA and the 'thioesterase I activity are directly correlated. Moreover, the polymer accumulated in the recombinant E. coli consisted mainly of 3-hydroxyoctanoate monomers. On the basis of our data, we propose an MCL PHA biosynthesis pathway scheme for recombinant E. coli JMU193, harboring PHA polymerase and 'thioesterase I, when grown on gluconate, which involves both de novo fatty acid synthesis and beta -oxidation.

* Corresponding author. Mailing address: Institute of Biotechnology, Swiss Federal Institute of Technology, ETH Zurich, Hoenggerberg HPT, CH-8093 Zurich, Switzerland. Phone: 41-1-633 3286. Fax: 41-1-633 1051. E-mail: bw{at}biotech.biol.ethz.ch.

Applied and Environmental Microbiology, February 1999, p. 540-548, Vol. 65, No. 2
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Sabirova, J. S., Ferrer, M., Lunsdorf, H., Wray, V., Kalscheuer, R., Steinbuchel, A., Timmis, K. N., Golyshin, P. N. (2006). Mutation in a "tesB-Like" Hydroxyacyl-Coenzyme A-Specific Thioesterase Gene Causes Hyperproduction of Extracellular Polyhydroxyalkanoates by Alcanivorax borkumensis SK2. J. Bacteriol. 188: 8452-8459 [Abstract] [Full Text]  
  • Zheng, Z., Gong, Q., Liu, T., Deng, Y., Chen, J.-C., Chen, G.-Q. (2004). Thioesterase II of Escherichia coli Plays an Important Role in 3-Hydroxydecanoic Acid Production. Appl. Environ. Microbiol. 70: 3807-3813 [Abstract] [Full Text]  
  • Nomura, C. T., Taguchi, K., Taguchi, S., Doi, Y. (2004). Coexpression of Genetically Engineered 3-Ketoacyl-ACP Synthase III (fabH) and Polyhydroxyalkanoate Synthase (phaC) Genes Leads to Short-Chain-Length-Medium-Chain-Length Polyhydroxyalkanoate Copolymer Production from Glucose in Escherichia coli JM109. Appl. Environ. Microbiol. 70: 999-1007 [Abstract] [Full Text]  
  • Rehm, B. H. A., Mitsky, T. A., Steinbuchel, A. (2001). Role of Fatty Acid De Novo Biosynthesis in Polyhydroxyalkanoic Acid (PHA) and Rhamnolipid Synthesis by Pseudomonads: Establishment of the Transacylase (PhaG)-Mediated Pathway for PHA Biosynthesis in Escherichia coli. Appl. Environ. Microbiol. 67: 3102-3109 [Abstract] [Full Text]  
  • Klinke, S., de Roo, G., Witholt, B., Kessler, B. (2000). Role of phaD in Accumulation of Medium-Chain-Length Poly(3-Hydroxyalkanoates) in Pseudomonas oleovorans. Appl. Environ. Microbiol. 66: 3705-3710 [Abstract] [Full Text]  
  • Fiedler, S., Steinbüchel, A., Rehm, B. H. A. (2000). PhaG-Mediated Synthesis of Poly(3-Hydroxyalkanoates) Consisting of Medium-Chain-Length Constituents from Nonrelated Carbon Sources in Recombinant Pseudomonas fragi. Appl. Environ. Microbiol. 66: 2117-2124 [Abstract] [Full Text]  
  • Klinke, S., Dauner, M., Scott, G., Kessler, B., Witholt, B. (2000). Inactivation of Isocitrate Lyase Leads to Increased Production of Medium-Chain-Length Poly(3-Hydroxyalkanoates) in Pseudomonas putida. Appl. Environ. Microbiol. 66: 909-913 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»