Biotechnology Letters

, Volume 36, Issue 11, pp 2257–2262 | Cite as

Metabolic engineering of Escherichia coli for poly(3-hydroxypropionate) production from glycerol and glucose

  • Qi Wang
  • Peng Yang
  • Mo Xian
  • Lu Feng
  • Jiming Wang
  • Guang Zhao
Original Research Paper
First Online:
Received:
Accepted:

Abstract

A new poly(3-hydroxypropionate) (P3HP) biosynthetic pathway employing β-alanine as an intermediate from an inexpensive carbon source was developed in recombinant Escherichia coli. After a series of systematic optimization, the genes for L-aspartate decarboxylase and its maturation factor (panD and panM, from E. coli), β-alanine-pyruvate transaminase (pp0596, from Pseudomonas putida), 3-hydroxy acid dehydrogenase and 3-hydroxypropionyl-CoA synthase (ydfG and prpE respectively, from E. coli), and polyhydroxyalkanoate synthase (phaC1, from Cupriavidus necator) were cloned and expressed in E. coli. Under shake-flask conditions, the recombinant strain produced 0.5 g P3HP l−1 from glycerol and glucose, up to 10.2 % of CDW. Though the content of P3HP was low, this pathway has some advantages over other reported pathways, such as being redox neutral, does not require any coenzyme, and can use a wide range of carbon sources.

Keywords

β-Alanine Inexpensive carbon sources Poly (3-hydroxypropionate) Recombinant Escherichia coli 
This is a preview of subscription content, log in to check access.

Notes

Acknowledgments

This research was financially supported by the 100-Talent Project of CAS (for GZ), Director Innovation Foundation of QIBEBT, CAS (Y112141105), National Natural Scientific Foundation of China (31200030, 21376255), Natural Scientific Foundation of Shandong Province (ZR2013EMZ002), and Qingdao Applied Basic Research Program (12-1-4-9-(5)-jch). We also acknowledge Dr. Birgit Alber (Ohio State University) for supplying the plasmid pKS1.

Supplementary material

10529_2014_1600_MOESM1_ESM.docx (235 kb)
Supplementary material 1 (DOCX 235 kb)

References

  1. Anderson AJ, Dawes EA (1990) Occurrence, metabolism, metabolic role, and industrial uses of bacterial polyhydroxyalkanoates. Microbiol Rev 54:450–472PubMedPubMedCentralGoogle Scholar
  2. Andreessen B, Steinbuchel A (2010) Biosynthesis and biodegradation of 3-hydroxypropionate-containing polyesters. Appl Environ Microbiol 76:4919–4925PubMedCrossRefPubMedCentralGoogle Scholar
  3. Andreessen B, Lange AB, Robenek H, Steinbuchel A (2010) Conversion of glycerol to poly(3-hydroxypropionate) in recombinant Escherichia coli. Appl Environ Microbiol 76:622–626PubMedCrossRefPubMedCentralGoogle Scholar
  4. Cao A, Kasuya K, Abe H, Doi Y, Inoue Y (1998) Studies on comonomer compositional distribution of the bacterial poly (3-hydroxybutyric acid-co-3-hydroxypropionic acid)s and crystal and thermal characteristics of their fractionated component copolyesters. Polymer 39(20):4801–4816CrossRefGoogle Scholar
  5. Chen GQ (2009) A microbial polyhydroxyalkanoates (PHA) based bio- and materials industry. Chem Soc Rev 38:2434–2446PubMedCrossRefGoogle Scholar
  6. Chowdhury EK, Akaishi Y, Nagata S, Misono H (2003) Cloning and overexpression of the 3-hydroxyisobutyrate dehydrogenase gene from pseudomonas putida E23. Biosci Biotechnol Biochem 67:438–441PubMedCrossRefGoogle Scholar
  7. Fujisawa H, Nagata S, Misono H (2003) Characterization of short-chain dehydrogenase/reductase homologues of Escherichia coli (YdfG) and Saccharomyces cerevisiae (YMR226C). BBA-Proteins Proteom 1645:89–94CrossRefGoogle Scholar
  8. Fukui T, Suzuki M, Tsuge T, Nakamura S (2009) Microbial synthesis of poly ((R)-3-hydroxybutyrate-co-3-hydroxypropionate) from unrelated carbon sources by engineered Cupriavidus necator. Biomacromolecules 10:700–706PubMedCrossRefGoogle Scholar
  9. Green PR, Kemper J, Schechtman L, Guo L, Satkowski M, Fiedler S, Steinbuchel A, Rehm BHA (2002) Formation of short chain length/medium chain length polyhydroxyalkanoate copolymers by fatty acid β-oxidationinhibited Ralstonia eutropha. Biomacromolecules 3:208–213PubMedCrossRefGoogle Scholar
  10. Hayaishi O, Nishizuka Y, Tatibana M, Takeshita M, Kuno S (1961) Enzymatic studies on the metabolism of β-Alanine. J Biol Chem 236:781–790PubMedGoogle Scholar
  11. Herter S, Fuchs G, Bacher A, Eisenreich W (2002) A bicyclic autotrophic CO2 fixation pathway in Chloroflexus aurantiacus. J Biol Chem 277:20277–20283PubMedCrossRefGoogle Scholar
  12. Liu Q, Luo G, Zhou XR, Chen GQ (2011) Biosynthesis of poly (3-hydroxydecanoate) and 3-hydroxydodecanoate dominating polyhydroxyalkanoates by β-oxidation pathway inhibited Pseudomonas putida. Metab Eng 13:11–17PubMedCrossRefGoogle Scholar
  13. Liu CS, Wang Q, Xian M, Ding YM, Zhao G (2013) Dissection of malonyl-coenzyme A reductase of Chloroflexus aurantiacus results in enzyme activity improvement. PLoS ONE 8(9):e75554PubMedCrossRefPubMedCentralGoogle Scholar
  14. Raj SM, Rathnasingh C, Jo JE, Park S (2008) Production of 3-hydroxypropionic acid from glycerol by a novel recombinant Escherichia coli BL21 strain. Process Biochem 43:1440–1446CrossRefGoogle Scholar
  15. Scott EM, Jakoby WB (1959) Soluble γ-aminobutyric-glutamic transaminase from Pseudomonas fluorescens. J Biol Chem 234:932–936PubMedGoogle Scholar
  16. Wang Q, Liu CS, Xian M, Zhang YG, Zhao G (2012) Biosynthetic pathway for poly(3-hydroxypropionate) in recombinant Escherichia coli. J Microbiol 50:693–697PubMedCrossRefGoogle Scholar
  17. Wang Q, Yang P, Liu CS, Xue YC, Xian M, Zhao G (2013) Biosynthesis of poly(3-hydroxypropionate) from glycerol by recombinant Escherichia coli. Bioresour Technol 131:548–551PubMedCrossRefGoogle Scholar
  18. Zhou Q, Shi ZY, Meng DC, Wu Q, Chen JC, Chen GQ (2011) Production of 3-hydroxypropionate homopolymer and poly (3-hydroxypropionate- co-4-hydroxybutyrate) copolymer by recombinant Escherichia coli. Metab Eng 13:777–785PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2014

Authors and Affiliations

  • Qi Wang
    • 1
    • 2
  • Peng Yang
    • 1
    • 3
  • Mo Xian
    • 1
  • Lu Feng
    • 1
  • Jiming Wang
    • 1
  • Guang Zhao
    • 1
  1. 1.CAS Key Laboratory of Biobased MaterialsQingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of SciencesQingdaoChina
  2. 2.University of Chinese Academy of SciencesBeijingChina
  3. 3.School of Biological EngineeringDalian Polytechnic UniversityDalianChina

Personalised recommendations