Abstract
Poly(3-hydroxypropionate) (P3HP) is a biodegradable and biocompatible thermoplastic. In this study, we engineered a P3HP biosynthetic pathway in recombinant Escherichia coli. The genes for malonyl-CoA reductase (mcr, from Chloroflexus aurantiacus), propionyl-CoA synthetase (prpE, from E. coli), and polyhydroxyalkanoate synthase (phaC1, from Ralstonia eutropha) were cloned and expressed in E. coli. The E. coli genes accABCD encoding acetyl-CoA carboxylase were used to channel the carbon into the P3HP pathway. Using glucose as a sole carbon source, the cell yield and P3HP content were 1.32 g/L and 0.98% (wt/wt [cell dry weight]), respectively. Although the yield is relatively low, our study shows the feasibility of engineering a P3HP biosynthetic pathway using a structurally unrelated carbon source in bacteria.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Andreeßen, B., Lange, A.B., Robenek, H., and Steinbüchel, A. 2010. Conversion of glycerol to poly(3-hydroxypropionate) in recombinant Escherichia coli. Appl. Environ. Microbiol. 76, 622–626.
Andreeßen, B. and Steinbüchel, A. 2010. Biosynthesis and biodegradation of 3-hydroxypropionate-containing polyesters. Appl. Environ. Microbiol. 76, 4919–4925.
Arai, Y., Cao, A., Yoshie, N., and Inoue, Y. 1999. Studies on comonomer compositional distribution and its effect on some physical properties of bacterial poly(3-hydroxybutyric acid-co-3-hydroxypropionic acid). Polym. Int. 48, 1219–1228.
Brandl, H., Gross, R.A., Lenz, R.W., and Fuller, R.C. 1988. Pseudomonas oleovorans as a source of poly(b-Hydroxyalkanoates) for potential applications as biodegradable polyesters. Appl. Environ. Microbiol. 54, 1977–1982.
Cao, Y.J., Jiang, X.L., Zhang, R.B., and Xian, M. 2011. Improved phloroglucinol production by metabolically engineered Escherichia coli. Appl. Microbiol. Biotechnol. 91, 1545–1552.
Chen, G.Q. 2009. A microbial polyhydroxyalkanoates (PHA) based bio- and materials industry. Chem. Soc. Rev. 38, 2434–2446.
Clarke, D.M. and Bragg, P.D. 1985. Cloning and expression of the transhydrogenase gene of Escherichia coli. J. Bacteriol. 162, 367–373.
Friedrich, B., Pohlmann, A., Fricke, W.F., Reinecke, F., Kusian, B., Liesegang, H., Cramm, R., Eitinger, T., Ewering, C., Potter, M., and et al. 2006. Genome sequence of the bioplastic-producing “Knallgas” bacterium Ralstonia eutropha H16. Nat. Biotechnol. 24, 1257–1262.
Fukui, T., Suzuki, M., Tsuge, T., and Nakamura, S. 2009. Microbial synthesis of poly((R)-3-hydroxybutyrate-co-3-hydroxypropionate) from unrelated carbon sources by engineered Cupriavidus necator. Biomacromolecules 10, 700–706.
Green, P.R., Kemper, J., Schechtman, L., Guo, L., Satkowski, M., Fiedler, S., Steinbüchel, A., and Rehm, B.H. 2002. Formation of short chain length/medium chain length polyhydroxyalkanoate copolymers by fatty acid beta-oxidation inhibited Ralstonia eutropha. Biomacromolecules 3, 208–213.
Gresham, T.L., Jansen, J.E., and Shaver, F.W. 1948. β-Proponlactone. I. Polymerization reactions. J. Am. Chem. Soc. 70, 998–999.
Horswill, A.R. and Escalante-Semerena, J.C. 1999. The prpE gene of Salmonella typhimurium LT2 encodes propionyl-CoA synthetase. Microbiology 145(Pt 6), 1381–1388.
Hugler, M., Menendez, C., Schagger, H., and Fuchs, G. 2002. Malonyl-coenzyme A reductase from Chloroflexus aurantiacus, a key enzyme of the 3-hydroxypropionate cycle for autotrophic CO2 fixation. J. Bacteriol. 184, 2404–2410.
Kang, C.K., Lee, H.S., and Kim, J.H. 1993. Accumulation of PHA and its copolyesters by Methylobacterium sp. KCTC 0048. Biotechnol. Lett. 15, 1017–1020.
Kroeger, J.K., Zarzycki, J., and Fuchs, G. 2011. A spectrophotometric assay for measuring acetyl-coenzyme A carboxylase. Anal. Biochem. 411, 100–105.
Lageveen, R.G., Huisman, G.W., Preusting, H., Ketelaar, P., Eggink, G., and Witholt, B. 1988. Formation of polyesters by Pseudomonas oleovorans — Effect of substrates on formation and composition of poly-(R)-3-hydroxyalkanoates and poly-(R)-3-hydroxyalkenoates. Appl. Environ. Microbiol. 54, 2924–2932.
Liu, Q., Luo, G., Zhou, X.R., and Chen, G.Q. 2011. Biosynthesis of poly(3-hydroxydecanoate) and 3-hydroxydodecanoate dominating polyhydroxyalkanoates by β-oxidation pathway inhibited Pseudomonas putida. Metab. Eng. 13, 11–17.
Nakamura, S., Kunioka, M., and Doi, Y. 1991. Biosynthesis and characterization of bacterial poly(3-hydroxybutyrate-co-3-hydroxypropionate). J. Macromol. Sci. Chem. A28, 15–24.
Ostle, A.G. and Holt, J.G. 1982. Nile blue A as a fluorescent stain for poly-β-hydroxybutyrate. Appl. Environ. Microbiol. 44, 238–241.
Poelarends, G.J., Johnson, W.H., Jr., Murzin, A.G., and Whitman, C.P. 2003. Mechanistic characterization of a bacterial malonate semialdehyde decarboxylase: identification of a new activity on the tautomerase superfamily. J. Biol. Chem. 278, 48674–48683.
Rathnasingh, C., Raj, S.M., Lee, Y., Catherine, C., Ashok, S., and Park, S. 2012. Production of 3-hydroxypropionic acid via malonyl-CoA pathway using recombinant Escherichia coli strains. J. Biotechnol. 157, 633–640.
Steinbüchel, A. and Lutke-Eversloh, T. 2003. Metabolic engineering and pathway construction for biotechnological production of relevant polyhydroxyalkanoates in microorganisms. Biochem. Eng. J. 16, 81–96.
Valentin, H.E., Mitsky, T.A., Mahadeo, D.A., Tran, M., and Gruys, K.J. 2000. Application of a propionyl coenzyme A synthetase for poly(3-hydroxypropionate-co-3-hydroxybutyrate) accumulation in recombinant Escherichia coli. Appl. Environ. Microbiol. 66, 5253–5258.
Wang, H.H., Li, X.T., and Chen, G.Q. 2009. Production and characterization of homopolymer polyhydroxyheptanoate (P3HHp) by a fadBA knockout mutant Pseudomonas putida KTOY06 derived from P. putida KT2442. Proc. Biochem. 44, 106–111.
Warnecke, T.E., Lynch, M.D., Karimpour-Fard, A., Sandoval, N., and Gill, R.T. 2008. A genomics approach to improve the analysis and design of strain selections. Metab. Eng. 10, 154–165.
Yamashita, M., Takemoto, Y., Ihara, E., and Yasuda, H. 1996. Organolanthanide-initiated living polymerizations of ɛ-caprolactone, δ-valerolactone, and β-propiolactone. Macromolecules 29, 1798–1806.
Zha, W., Rubin-Pitel, S.B., Shao, Z., and Zhao, H. 2009. Improving cellular malonyl-CoA level in Escherichia coli via metabolic engineering. Metab. Eng. 11, 192–198.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Wang, Q., Liu, C., Xian, M. et al. Biosynthetic pathway for poly(3-Hydroxypropionate) in recombinant Escherichia coli . J Microbiol. 50, 693–697 (2012). https://doi.org/10.1007/s12275-012-2234-y
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12275-012-2234-y