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Applied and Environmental Microbiology, January 2006, p. 536-543, Vol. 72, No. 1
0099-2240/06/$08.00+0     doi:10.1128/AEM.72.1.536-543.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Engineering the Monomer Composition of Polyhydroxyalkanoates Synthesized in Saccharomyces cerevisiae

Bo Zhang, Ross Carlson,^, and Friedrich Srienc^*

Department of Chemical Engineering and Materials Science, University of Minnesota, 151 Amundson Hall, 421 Washington Avenue SE, Minneapolis, Minnesota 55455, and BioTechnology Institute, University of Minnesota, 240 Gortner Labs, 1479 Gortner Avenue, St. Paul, Minnesota 55108

Received 9 June 2005/ Accepted 22 October 2005

Polyhydroxyalkanoates (PHAs) have received considerable interest as renewable-resource-based, biodegradable, and biocompatible plastics with a wide range of potential applications. We have engineered the synthesis of PHA polymers composed of monomers ranging from 4 to 14 carbon atoms in either the cytosol or the peroxisome of Saccharomyces cerevisiae by harnessing intermediates of fatty acid metabolism. Cytosolic PHA production was supported by establishing in the cytosol critical ß-oxidation chemistries which are found natively in peroxisomes. This platform was utilized to supply medium-chain (C₆ to C₁₄) PHA precursors from both fatty acid degradation and synthesis to a cytosolically expressed medium-chain-length (mcl) polymerase from Pseudomonas oleovorans. Synthesis of short-chain-length PHAs (scl-PHAs) was established in the peroxisome of a wild-type yeast strain by targeting the Ralstonia eutropha scl polymerase to the peroxisome. This strain, harboring a peroxisomally targeted scl-PHA synthase, accumulated PHA up to approximately 7% of its cell dry weight. These results indicate (i) that S. cerevisiae expressing a cytosolic mcl-PHA polymerase or a peroxisomal scl-PHA synthase can use the 3-hydroxyacyl coenzyme A intermediates from fatty acid metabolism to synthesize PHAs and (ii) that fatty acid degradation is also possible in the cytosol as ß-oxidation might not be confined only to the peroxisomes. Polymers of even-numbered, odd-numbered, or a combination of even- and odd-numbered monomers can be controlled by feeding the appropriate substrates. This ability should permit the rational design and synthesis of polymers with desired material properties.

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* Corresponding author. Mailing address: BioTechnology Institute, University of Minnesota, 240 Gortner Labs, 1479 Gortner Avenue, St. Paul, MN 55108. Phone: (612) 624-9776. Fax: (612) 625-1700. E-mail: srienc{at}umn.edu

Present address: Center for Biofilm Engineering, Department of Chemical and Biological Engineering, Montana State University, Bozeman, MT 59717.

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Applied and Environmental Microbiology, January 2006, p. 536-543, Vol. 72, No. 1
0099-2240/06/$08.00+0     doi:10.1128/AEM.72.1.536-543.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

This article has been cited by other articles:

• Nevoigt, E. (2008). Progress in Metabolic Engineering of Saccharomyces cerevisiae. Microbiol. Mol. Biol. Rev. 72: 379-412 [Abstract] [Full Text]