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Microbial production of polyhydroxyalkanoate block copolymer by recombinant Pseudomonas putida

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Abstract

Polyhydroxyalkanoate (PHA) synthesis genes phaPCJ Ac cloned from Aeromonas caviae were transformed into Pseudomonas putida KTOY06ΔC, a mutant of P. putida KT2442, resulting in the ability of the recombinant P. putida KTOY06ΔC (phaPCJ A.c ) to produce a short-chain-length and medium-chain-length PHA block copolymer consisting of poly-3-hydroxybutyrate (PHB) as one block and random copolymer of 3-hydroxyvalerate (3HV) and 3-hydroxyheptanoate (3HHp) as another block. The novel block polymer was studied by differential scanning calorimetry (DSC), nuclear magnetic resonance, and rheology measurements. DSC studies showed the polymer to possess two glass transition temperatures (T g), one melting temperature (T m) and one cool crystallization temperature (T c). Rheology studies clearly indicated a polymer chain re-arrangement in the copolymer; these studies confirmed the polymer to be a block copolymer, with over 70 mol% homopolymer (PHB) of 3-hydroxybutyrate (3HB) as one block and around 30 mol% random copolymers of 3HV and 3HHp as the second block. The block copolymer was shown to have the highest tensile strength and Young’s modulus compared with a random copolymer with similar ratio and a blend of homopolymers PHB and PHVHHp with similar ratio. Compared with other commercially available PHA including PHB, PHBV, PHBHHx, and P3HB4HB, the short-chain- and medium-chain-length block copolymer PHB-b-PHVHHp showed differences in terms of mechanical properties and should draw more attentions from the PHA research community.

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Acknowledgement

Thanks to Mr. Honghui Wang for kindly making the homopolymers PHV and PHHp for this study. And also thanks to Dr. Shao Ping Ouyang of Tsinghua University for kindly donating P. putida KTOY06ΔC (phaPCJ A.c ). This research was financially supported by the Li Ka Shing Foundation, the National High Tech 863 Grants (project 2010AA101607), and the 973 Basic Research Fund (grant no. 2007CB707804).

Author information

Correspondence to Guo-Qiang Chen.

Additional information

Shi Yan Li, Cui Ling Dong, and Shen Yu Wang have contributed equally to this paper.

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Li, S.Y., Dong, C.L., Wang, S.Y. et al. Microbial production of polyhydroxyalkanoate block copolymer by recombinant Pseudomonas putida . Appl Microbiol Biotechnol 90, 659–669 (2011) doi:10.1007/s00253-010-3069-2

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Keywords

  • PHB
  • Polyhydroxyalkanoates
  • Block copolymer
  • Pseudomonas putida
  • 3-Hydroxybutyrate
  • 3-Hydroxyvalerate
  • 3-Hydroxyheptanoate