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 Fiedler, S.
Right arrow Articles by Rehm, B. H. A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Fiedler, S.
Right arrow Articles by Rehm, B. H. A.
Agricola
Right arrow Articles by Fiedler, S.
Right arrow Articles by Rehm, B. H. A.

 Previous Article  |  Next Article 

Applied and Environmental Microbiology, May 2000, p. 2117-2124, Vol. 66, No. 5
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

PhaG-Mediated Synthesis of Poly(3-Hydroxyalkanoates) Consisting of Medium-Chain-Length Constituents from Nonrelated Carbon Sources in Recombinant Pseudomonas fragi

Silke Fiedler, Alexander Steinbüchel, and Bernd H. A. Rehm*

Institut für Mikrobiologie, Westfälische Wilhelms-Universität Münster, D-48149 Münster, Germany

Received 30 November 1999/Accepted 5 March 2000

Recently, a new metabolic link between fatty acid de novo biosynthesis and biosynthesis of poly(3-hydroxy-alkanoate) consisting of medium-chain-length constituents (C6 to C14) (PHAMCL), catalyzed by the 3-hydroxydecanoyl-[acyl-carrier-protein]:CoA transacylase (PhaG), has been identified in Pseudomonas putida (B. H. A. Rehm, N. Krüger, and A. Steinbüchel, J. Biol. Chem. 273:24044-24051, 1998). To establish this PHA-biosynthetic pathway in a non-PHA-accumulating bacterium, we functionally coexpressed phaC1 (encoding PHA synthase 1) from Pseudomonas aeruginosa and phaG (encoding the transacylase) from P. putida in Pseudomonas fragi. The recombinant strains of P. fragi were cultivated on gluconate as the sole carbon source, and PHA accumulation to about 14% of the total cellular dry weight was achieved. The respective polyester was isolated, and GPC analysis revealed a weight average molar mass of about 130,000 g mol-1 and a polydispersity of 2.2. The PHA was composed mainly (60 mol%) of 3-hydroxydecanoate. These data strongly suggested that functional expression of phaC1 and phaG established a new pathway for PHAMCL biosynthesis from nonrelated carbon sources in P. fragi. When fatty acids were used as the carbon source, no PHA accumulation was observed in PHA synthase-expressing P. fragi, whereas application of the beta -oxidation inhibitor acrylic acid mediated PHAMCL accumulation. The substrate for the PHA synthase PhaC1 is therefore presumably directly provided through the enzymatic activity of the transacylase PhaG by the conversion of (R)-3-hydroxydecanoyl-ACP to (R)-3-hydroxydecanoyl-CoA when the organism is cultivated on gluconate. Here we demonstrate for the first time the establishment of PHAMCL synthesis from nonrelated carbon sources in a non-PHA-accumulating bacterium, employing fatty acid de novo biosynthesis and the enzymes PhaG (a transacylase) and PhaC1 (a PHA synthase).

* Corresponding author. Mailing address: Institut für Mikrobiologie, Westfälische Wilhelms-Universität Münster, Corrensstrasse 3, D-48149 Münster, Germany. Phone: (49) 251 833 9848. Fax: (49) 251 833 8388. E-mail: rehm{at}uni-muenster.de.

Applied and Environmental Microbiology, May 2000, p. 2117-2124, Vol. 66, No. 5
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • O'Leary, N. D., O'Connor, K. E., Ward, P., Goff, M., Dobson, A. D. W. (2005). Genetic Characterization of Accumulation of Polyhydroxyalkanoate from Styrene in Pseudomonas putida CA-3. Appl. Environ. Microbiol. 71: 4380-4387 [Abstract] [Full Text]  
  • Ward, P. G., de Roo, G., O'Connor, K. E. (2005). Accumulation of Polyhydroxyalkanoate from Styrene and Phenylacetic Acid by Pseudomonas putida CA-3. Appl. Environ. Microbiol. 71: 2046-2052 [Abstract] [Full Text]  
  • Pham, T. H., Webb, J. S., Rehm, B. H. A. (2004). The role of polyhydroxyalkanoate biosynthesis by Pseudomonas aeruginosa in rhamnolipid and alginate production as well as stress tolerance and biofilm formation. Microbiology 150: 3405-3413 [Abstract] [Full Text]  
  • Hoffmann, N., Amara, A. A., Beermann, Br. B., Qi, Q., Hinz, H.-J., Rehm, B. H. A. (2002). Biochemical Characterization of the Pseudomonas putida 3-Hydroxyacyl ACP:CoA Transacylase, Which Diverts Intermediates of Fatty Acid de Novo Biosynthesis. J. Biol. Chem. 277: 42926-42936 [Abstract] [Full Text]  
  • Potter, M., Madkour, M. H., Mayer, F., Steinbuchel, A. (2002). Regulation of phasin expression and polyhydroxyalkanoate (PHA) granule formation in Ralstonia eutropha H16. Microbiology 148: 2413-2426 [Abstract] [Full Text]  
  • Qi, Q., Rehm, B. H. A. (2001). Polyhydroxybutyrate biosynthesis in Caulobacter crescentus: molecular characterization of the polyhydroxybutyrate synthase. Microbiology 147: 3353-3358 [Abstract] [Full Text]  
  • Lee, H.-J., Choi, M. H., Kim, T.-U., Yoon, S. C. (2001). Accumulation of Polyhydroxyalkanoic Acid Containing Large Amounts of Unsaturated Monomers in Pseudomonas fluorescens BM07 Utilizing Saccharides and Its Inhibition by 2-Bromooctanoic Acid. Appl. Environ. Microbiol. 67: 4963-4974 [Abstract] [Full Text]  
  • Poirier, Y., Erard, N., Petetot, J. M.-C. (2001). Synthesis of Polyhydroxyalkanoate in the Peroxisome of Saccharomyces cerevisiae by Using Intermediates of Fatty Acid beta -Oxidation. Appl. Environ. Microbiol. 67: 5254-5260 [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]  


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