Engineering the Monomer Composition of Polyhydroxyalkanoates Synthesized in Saccharomyces Cerevisiae

Overview

Journal Appl Environ Microbiol

Specialties Environmental Health
Microbiology

Date 2006 Jan 5

PMID 16391089

Citations 12

Authors

Bo Zhang

Ross Carlson

Friedrich Srienc

Affiliations

Soon will be listed here.

Abstract

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 beta-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 beta-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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References

Hahn J, Eschenlauer A, Sleytr U, Somers D, Srienc F . Peroxisomes as sites for synthesis of polyhydroxyalkanoates in transgenic plants. Biotechnol Prog. 1999; 15(6):1053-7. DOI: 10.1021/bp990118n. View

Ashrafi K, Farazi T, Gordon J . A role for Saccharomyces cerevisiae fatty acid activation protein 4 in regulating protein N-myristoylation during entry into stationary phase. J Biol Chem. 1998; 273(40):25864-74. DOI: 10.1074/jbc.273.40.25864. View

Kessler B, Weusthuis R, Witholt B, Eggink G . Production of microbial polyesters: fermentation and downstream processes. Adv Biochem Eng Biotechnol. 2001; 71:159-82. DOI: 10.1007/3-540-40021-4_5. View

van Roermund C, Drissen R, van den Berg M, IJlst L, Hettema E, Tabak H . Identification of a peroxisomal ATP carrier required for medium-chain fatty acid beta-oxidation and normal peroxisome proliferation in Saccharomyces cerevisiae. Mol Cell Biol. 2001; 21(13):4321-9. PMC: 87092. DOI: 10.1128/MCB.21.13.4321-4329.2001. View

Faergeman N, Black P, Zhao X, Knudsen J, DiRusso C . The Acyl-CoA synthetases encoded within FAA1 and FAA4 in Saccharomyces cerevisiae function as components of the fatty acid transport system linking import, activation, and intracellular Utilization. J Biol Chem. 2001; 276(40):37051-9. DOI: 10.1074/jbc.M100884200. View

Poirier Y, Erard N, Petetot J . Synthesis of polyhydroxyalkanoate in the peroxisome of Saccharomyces cerevisiae by using intermediates of fatty acid beta-oxidation. Appl Environ Microbiol. 2001; 67(11):5254-60. PMC: 93298. DOI: 10.1128/AEM.67.11.5254-5260.2001. View

Poirier Y, Erard N, Petetot J . Synthesis of polyhydroxyalkanoate in the peroxisome of Pichia pastoris. FEMS Microbiol Lett. 2002; 207(1):97-102. DOI: 10.1111/j.1574-6968.2002.tb11035.x. View

Sherman F . Getting started with yeast. Methods Enzymol. 2002; 350:3-41. DOI: 10.1016/s0076-6879(02)50954-x. View

Klein A, van den Berg M, Bottger G, Tabak H, Distel B . Saccharomyces cerevisiae acyl-CoA oxidase follows a novel, non-PTS1, import pathway into peroxisomes that is dependent on Pex5p. J Biol Chem. 2002; 277(28):25011-9. DOI: 10.1074/jbc.M203254200. View

10.

Carlson R, Fell D, Srienc F . Metabolic pathway analysis of a recombinant yeast for rational strain development. Biotechnol Bioeng. 2002; 79(2):121-34. DOI: 10.1002/bit.10305. View

11.

Marchesini S, Poirier Y . Futile cycling of intermediates of fatty acid biosynthesis toward peroxisomal beta-oxidation in Saccharomyces cerevisiae. J Biol Chem. 2003; 278(35):32596-601. DOI: 10.1074/jbc.M305574200. View

12.

De Oliveira V, Maeda I, Delessert S, Poirier Y . Increasing the carbon flux toward synthesis of short-chain-length--medium-chain-length polyhydroxyalkanoate in the peroxisome of Saccharomyces cerevisiae through modification of the beta-oxidation cycle. Appl Environ Microbiol. 2004; 70(9):5685-7. PMC: 520895. DOI: 10.1128/AEM.70.9.5685-5687.2004. View

13.

Erdmann R, Veenhuis M, Mertens D, Kunau W . Isolation of peroxisome-deficient mutants of Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1989; 86(14):5419-23. PMC: 297634. DOI: 10.1073/pnas.86.14.5419. View

14.

Huisman G, Wonink E, Meima R, Kazemier B, Terpstra P, Witholt B . Metabolism of poly(3-hydroxyalkanoates) (PHAs) by Pseudomonas oleovorans. Identification and sequences of genes and function of the encoded proteins in the synthesis and degradation of PHA. J Biol Chem. 1991; 266(4):2191-8. View

15.

van der Leij I, van den Berg M, Boot R, Franse M, Distel B, Tabak H . Isolation of peroxisome assembly mutants from Saccharomyces cerevisiae with different morphologies using a novel positive selection procedure. J Cell Biol. 1992; 119(1):153-62. PMC: 2289622. DOI: 10.1083/jcb.119.1.153. View

16.

Soni R, Carmichael J, Murray J . Parameters affecting lithium acetate-mediated transformation of Saccharomyces cerevisiae and development of a rapid and simplified procedure. Curr Genet. 1993; 24(5):455-9. DOI: 10.1007/BF00351857. View

17.

Knoll L, Johnson D, Gordon J . Biochemical studies of three Saccharomyces cerevisiae acyl-CoA synthetases, Faa1p, Faa2p, and Faa3p. J Biol Chem. 1994; 269(23):16348-56. View

18.

Leaf T, Peterson M, Stoup S, Somers D, Srienc F . Saccharomyces cerevisiae expressing bacterial polyhydroxybutyrate synthase produces poly-3-hydroxybutyrate. Microbiology (Reading). 1996; 142 ( Pt 5):1169-1180. DOI: 10.1099/13500872-142-5-1169. View

19.

Hettema E, van Roermund C, Distel B, van den Berg M, Vilela C, Rodrigues-Pousada C . The ABC transporter proteins Pat1 and Pat2 are required for import of long-chain fatty acids into peroxisomes of Saccharomyces cerevisiae. EMBO J. 1996; 15(15):3813-22. PMC: 452064. View

20.

Elgersma Y, Vos A, van den Berg M, van Roermund C, van der Sluijs P, Distel B . Analysis of the carboxyl-terminal peroxisomal targeting signal 1 in a homologous context in Saccharomyces cerevisiae. J Biol Chem. 1996; 271(42):26375-82. DOI: 10.1074/jbc.271.42.26375. View