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 York, G. M.
Right arrow Articles by Sinskey, A. J.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by York, G. M.
Right arrow Articles by Sinskey, A. J.

 Previous Article  |  Next Article 

Journal of Bacteriology, July 2003, p. 3788-3794, Vol. 185, No. 13
0021-9193/03/$08.00+0     DOI: 10.1128/JB.185.13.3788-3794.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Ralstonia eutropha H16 Encodes Two and Possibly Three Intracellular Poly[D-(-)-3-Hydroxybutyrate] Depolymerase Genes

Gregory M. York,1,{dagger} Joachim Lupberger,1,{ddagger} Jiamin Tian,2 Adam G. Lawrence,1 JoAnne Stubbe,1,2* and Anthony J. Sinskey1*

Departments of Biology,1 Chemistry, Massachusetts Institute of Technology, Cambridge, Massachusetts 021392

Received 14 January 2003/ Accepted 16 April 2003

Intracellular poly[D-(-)-3-hydroxybutyrate] (PHB) depolymerases degrade PHB granules to oligomers and monomers of 3-hydroxybutyric acid. Recently an intracellular PHB depolymerase gene (phaZ1) from Ralstonia eutropha was identified. We now report identification of candidate PHB depolymerase genes from R. eutropha, namely, phaZ2 and phaZ3, and their characterization in vivo. phaZ1 was used to identify two candidate depolymerase genes in the genome of Ralstonia metallidurans. phaZ1 and these genes were then used to design degenerate primers. These primers and PCR methods on the R. eutropha genome were used to identify two new candidate depolymerase genes in R. eutropha: phaZ2 and phaZ3. Inverse PCR methods were used to obtain the complete sequence of phaZ3, and library screening was used to obtain the complete sequence of phaZ2. PhaZ1, PhaZ2, and PhaZ3 share ~30% sequence identity. The function of PhaZ2 and PhaZ3 was examined by generating R. eutropha H16 deletion strains ({Delta}phaZ1, {Delta}phaZ2, {Delta}phaZ3, {Delta}phaZ1{Delta}phaZ2, {Delta}phaZ1{Delta}phaZ3, {Delta}phaZ2{Delta}phaZ3, and {Delta}phaZ1{Delta}phaZ2{Delta}phaZ3). These strains were analyzed for PHB production and utilization under two sets of conditions. When cells were grown in rich medium, PhaZ1 was sufficient to account for intracellular PHB degradation. When cells that had accumulated ~80% (cell dry weight) PHB were subjected to PHB utilization conditions, PhaZ1 and PhaZ2 were sufficient to account for PHB degradation. PhaZ2 is thus suggested to be an intracellular depolymerase. The role of PhaZ3 remains to be established.

* Corresponding author. Mailing address for JoAnne Stubbe: Department of Chemistry, Bldg. 68-438, Massachusetts Institute of Technology, 77 Massachusetts Ave., Cambridge, MA 02139. Phone: (617) 253-1814. Fax: (617) 258-7247. E-mail: stubbe{at}mit.edu. Mailing address for Anthony Sinskey: Department of Biology, Bldg. 68-370, Massachusetts Institute of Technology, 77 Massachusetts Ave., Cambridge, MA 02139. Phone: (617) 253-6721. Fax: (617) 253-8550. E-mail: asinskey{at}mit.edu.

{dagger} Present address: USB Corporation, Cleveland, OH 44128.

{ddagger} Present address: Feldmoos 5, D-88273 Fronreute, Germany.

Journal of Bacteriology, July 2003, p. 3788-3794, Vol. 185, No. 13
0021-9193/03/$08.00+0     DOI: 10.1128/JB.185.13.3788-3794.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Jendrossek, D. (2009). Polyhydroxyalkanoate Granules Are Complex Subcellular Organelles (Carbonosomes). J. Bacteriol. 191: 3195-3202 [Full Text]  
  • Raberg, M., Reinecke, F., Reichelt, R., Malkus, U., Konig, S., Potter, M., Fricke, W. F., Pohlmann, A., Voigt, B., Hecker, M., Friedrich, B., Bowien, B., Steinbuchel, A. (2008). Ralstonia eutropha H16 Flagellation Changes According to Nutrient Supply and State of Poly(3-Hydroxybutyrate) Accumulation. Appl. Environ. Microbiol. 74: 4477-4490 [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]  
  • Uchino, K., Saito, T., Gebauer, B., Jendrossek, D. (2007). Isolated Poly(3-Hydroxybutyrate) (PHB) Granules Are Complex Bacterial Organelles Catalyzing Formation of PHB from Acetyl Coenzyme A (CoA) and Degradation of PHB to Acetyl-CoA. J. Bacteriol. 189: 8250-8256 [Abstract] [Full Text]  
  • de Eugenio, L. I., Garcia, P., Luengo, J. M., Sanz, J. M., Roman, J. S., Garcia, J. L., Prieto, M. A. (2007). Biochemical Evidence That phaZ Gene Encodes a Specific Intracellular Medium Chain Length Polyhydroxyalkanoate Depolymerase in Pseudomonas putida KT2442: CHARACTERIZATION OF A PARADIGMATIC ENZYME. J. Biol. Chem. 282: 4951-4962 [Abstract] [Full Text]  
  • Tseng, C.-L., Chen, H.-J., Shaw, G.-C. (2006). Identification and Characterization of the Bacillus thuringiensis phaZ Gene, Encoding New Intracellular Poly-3-Hydroxybutyrate Depolymerase. J. Bacteriol. 188: 7592-7599 [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]  
  • Abe, T., Kobayashi, T., Saito, T. (2005). Properties of a Novel Intracellular Poly(3-Hydroxybutyrate) Depolymerase with High Specific Activity (PhaZd) in Wautersia eutropha H16. J. Bacteriol. 187: 6982-6990 [Abstract] [Full Text]  
  • Kobayashi, T., Uchino, K., Abe, T., Yamazaki, Y., Saito, T. (2005). Novel Intracellular 3-Hydroxybutyrate-Oligomer Hydrolase in Wautersia eutropha H16. J. Bacteriol. 187: 5129-5135 [Abstract] [Full Text]  
  • Waltermann, M., Steinbuchel, A. (2005). Neutral Lipid Bodies in Prokaryotes: Recent Insights into Structure, Formation, and Relationship to Eukaryotic Lipid Depots. J. Bacteriol. 187: 3607-3619 [Full Text]  
  • Tian, J., Sinskey, A. J., Stubbe, J. (2005). Kinetic Studies of Polyhydroxybutyrate Granule Formation in Wautersia eutropha H16 by Transmission Electron Microscopy. J. Bacteriol. 187: 3814-3824 [Abstract] [Full Text]  
  • Tian, J., He, A., Lawrence, A. G., Liu, P., Watson, N., Sinskey, A. J., Stubbe, J. (2005). Analysis of Transient Polyhydroxybutyrate Production in Wautersia eutropha H16 by Quantitative Western Analysis and Transmission Electron Microscopy. J. Bacteriol. 187: 3825-3832 [Abstract] [Full Text]  
  • Handrick, R., Reinhardt, S., Kimmig, P., Jendrossek, D. (2004). The "Intracellular" Poly(3-Hydroxybutyrate) (PHB) Depolymerase of Rhodospirillum rubrum Is a Periplasm-Located Protein with Specificity for Native PHB and with Structural Similarity to Extracellular PHB Depolymerases. J. Bacteriol. 186: 7243-7253 [Abstract] [Full Text]  
  • Potter, M., Muller, H., Reinecke, F., Wieczorek, R., Fricke, F., Bowien, B., Friedrich, B., Steinbuchel, A. (2004). The complex structure of polyhydroxybutyrate (PHB) granules: four orthologous and paralogous phasins occur in Ralstonia eutropha. Microbiology 150: 2301-2311 [Abstract] [Full Text]  
  • Handrick, R., Reinhardt, S., Schultheiss, D., Reichart, T., Schuler, D., Jendrossek, V., Jendrossek, D. (2004). Unraveling the Function of the Rhodospirillum rubrum Activator of Polyhydroxybutyrate (PHB) Degradation: the Activator Is a PHB-Granule-Bound Protein (Phasin). J. Bacteriol. 186: 2466-2475 [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»