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Microbial production of 4-hydroxybutyrate, poly-4-hydroxybutyrate, and poly(3-hydroxybutyrate-co-4-hydroxybutyrate) by recombinant microorganisms

  • Applied Genetics and Molecular Biotechnology
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Abstract

4-Hydroxybutyrate (4HB) was produced by Aeromonas hydrophila 4AK4, Escherichia coli S17-1, or Pseudomonas putida KT2442 harboring 1,3-propanediol dehydrogenase gene dhaT and aldehyde dehydrogenase gene aldD from P. putida KT2442 which are capable of transforming 1,4-butanediol (1,4-BD) to 4HB. 4HB containing fermentation broth was used for production of homopolymer poly-4-hydroxybutyrate [P(4HB)] and copolymers poly(3-hydroxybutyrate-co-4-hydroxybutyrate) [P(3HB-4HB)]. Recombinant A. hydrophila 4AK4 harboring plasmid pZL-dhaT-aldD containing dhaT and aldD was the most effective 4HB producer, achieving approximately 4 g/l 4HB from 10 g/l 1,4-BD after 48 h of incubation. The strain produced over 10 g/l 4HB from 20 g/l 1,4-BD after 52 h of cultivation in a 6-L fermenter. Recombinant E. coli S17-1 grown on 4HB containing fermentation broth was found to accumulate 83 wt.% of intracellular P(4HB) in shake flask study. Recombinant Ralstonia eutropha H16 grew to over 6 g/l cell dry weight containing 49 wt.% P(3HB-13%4HB) after 72 h.

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Acknowledgments

This research was supported by the National High Technology Research and Development Program of China (863 Program) Grant no. 2006AA02Z242 and 2006AA020104, the Natural Science Foundation of China Grant no. 30570024/C010103, and the Major State Basic Research Development Program of China (973 Program) Grant no. 2007CB707804. We are also very grateful to Professor Shuishan SONG (Hebei Institute of Biology, Shijiazhuang, Hebei, 050081, China) for his kind donation of plasmid pKSSE5.3.

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Authors and Affiliations

  1. Department of Biological Sciences and Biotechnology, Tsinghua University, Beijing, 100084, China

    Lei Zhang, Zhen-Yu Shi, Qiong Wu & Guo-Qiang Chen

  2. Multidisciplinary Research Center, Shantou University, Shantou, 515063, China

    Guo-Qiang Chen

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  1. Lei Zhang
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  2. Zhen-Yu Shi
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Correspondence to Qiong Wu or Guo-Qiang Chen.

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Zhang, L., Shi, ZY., Wu, Q. et al. Microbial production of 4-hydroxybutyrate, poly-4-hydroxybutyrate, and poly(3-hydroxybutyrate-co-4-hydroxybutyrate) by recombinant microorganisms. Appl Microbiol Biotechnol 84, 909–916 (2009). https://doi.org/10.1007/s00253-009-2023-7

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