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Biosynthesis of polyhydroxyalkanoate homopolymers by Pseudomonas putida

  • Applied Genetics and Molecular Biotechnology
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Abstract

Pseudomonas putida KT2442 has been a well-studied producer of medium-chain-length (mcl) polyhydroxyalkanoate (PHA) copolymers containing C6 ~ C14 monomer units. A mutant was constructed from P. putida KT2442 by deleting its phaG gene encoding R-3-hydroxyacyl-ACP-CoA transacylase and several other β-oxidation related genes including fadB, fadA, fadB2x, and fadAx. This mutant termed P. putida KTHH03 synthesized mcl homopolymers including poly(3-hydroxyhexanoate) (PHHx) and poly(3-hydroxyheptanoate) (PHHp), together with a near homopolymer poly(3-hydroxyoctanoate-co-2 mol% 3-hydroxyhexanoate) (PHO*) in presence of hexanoate, heptanoate, and octanoate, respectively. When deleted with its mcl PHA synthase genes phaC1 and phaC2, the recombinant mutant termed P. putida KTHH08 harboring pZWJ4-31 containing PHA synthesis operon phaPCJ from Aeromonas hydrophila 4AK4 accumulated homopolymer poly(3-hydroxyvalerate) (PHV) when valerate was used as carbon source. The phaC deleted recombinant mutant termed P. putida KTHH06 harboring pBHH01 holding PHA synthase PhbC from Ralstonia eutropha produced homopolymers poly(3-hydroxybutyrate) (PHB) and poly(4-hydroxybutyrate) using γ-butyrolactone was added as precursor. All the homopolymers were physically characterized. Their weight average molecular weights ranged from 1.8 × 105 to 1.6 × 106, their thermal stability changed with side chain lengths. The derivatives of P. putida KT2442 have been developed into a platform for production of various PHA homopolymers.

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Acknowledgements

This research was supported by Li Ka Shing Foundation and National High Tech 863 Grant (project no. 2006AA02Z242), as well as the State Basic Science Foundation 973 (2007CB707804). We thank Dr. Ouyang SP of Tsinghua University for kindly donating P. putida KTOY06, KTOY08, and plasmid pSPK02.We also thank Dr. Haimu Ye, Mr. Linping Wu, Liang Wang, and Hao Ni for many other forms of help they gave. In addition, Mr. Shenyu Wang, Guangyao Li, Wenfeng Ou, and Jueyu Pan provided assistances in the form of the use of some facilities for cell growth and physical characterization.

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Correspondence to Guo-Qiang Chen.

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Hong-hui Wang and Xin-rong Zhou contributed equally to this paper.

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Wang, Hh., Zhou, Xr., Liu, Q. et al. Biosynthesis of polyhydroxyalkanoate homopolymers by Pseudomonas putida . Appl Microbiol Biotechnol 89, 1497–1507 (2011). https://doi.org/10.1007/s00253-010-2964-x

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  • DOI: https://doi.org/10.1007/s00253-010-2964-x

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