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

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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References

  1. Anderson AJ, Dawes EA (1990) Occurrence, metabolism, metabolic role, and industrial uses of bacterial polyhydroxyalkanoates. Microbiol Rev 54:450–472

  2. Barbirato F, Larguier A, Conte T, Astruc S, Bories A (1997) Sensitivity to pH, product inhibition, and inhibition by NAD+ of 1, 3-propanediol dehydrogenase purified from Enterobacter agglomerans CNCM 1210. Arch Microbiol 168:160–163

  3. Chen GQ, Wu Q (2005) Microbial production and applications of chiral hydroxyalkanoates. Appl Microbiol Biotechnol 67:592–599

  4. Chen JY, Liu T, Zheng Z, Chen JC, Chen GQ (2004) Polyhydroxyalkanoate synthases PhaC1 and PhaC2 from Pseudomonas stutzeri 1317 had different substrate specificities. FEMS Microbiol Lett 234:231–237

  5. Choi MH, Yoon SC, Lenz RW (1999) Production of poly(3-hydroxybutyric acid-co-4-hydroxybutyric acid) and poly(4-hydroxybutyric acid) without subsequent degradation by Hydrogenophaga pseudoflava. Appl Environ Microbiol 65:1570–1577

  6. Daniel R, Boenigk R, Gottschalk G (1995) Purification of 1, 3-propanediol dehydrogenase from Citrobacter freundii and cloning, sequencing, and overexpression of the corresponding gene in Escherichia coli. J Bacteriol 177:2151–2156

  7. Drasbek KR, Christensen J, Jensen K (2006) Gamma-hydroxybutyrate—a drug of abuse. Acta Neurol Scand 114:145–156

  8. Efe C, Straathof AJJ, van der Wielen LAM (2008) Options for biochemical production of 4-hydroxybutyrate and its lactone as a substitute for petrochemical production. Biotechnol Bioeng 99:1392–1406

  9. Friedrich B, Hogrefe C, Schlegel HG (1981) Naturally occurring genetic transfer of hydrogen-oxidizing ability between strains of Alcaligenes eutrophus. J Bacteriol 147:198–205

  10. Gao HJ, Wu Q, Chen GQ (2002) Enhanced production of D-(−)-3-hydroxybutyric acid by recombinant Escherichia coli. FEMS Microbiol Lett 213:59–65

  11. Gerhardt A, Cinkaya I, Linder D, Huisman G, Buckel W (2000) Fermentation of 4-aminobutyrate by Clostridium aminobutyricum: cloning of two genes involved in the formation and dehydration of 4-hydroxybutyryl-CoA. Arch Microbiol 174:189–199

  12. Green LS, Li YZ, Emerich DW, Bergersen FJ, Day DA (2000) Catabolism of alpha-ketoglutarate by a sucA mutant of Bradyrhizobium japonicum: evidence for an alternative tricarboxylic acid cycle. J Bacteriol 182:2838–2844

  13. Hein S, Söhling B, Gottschalk G, Steinbüchel A (1997) Biosynthesis of poly(4-hydroxybutyric acid) by recombinant strains of Escherichia coli. FEMS Microbiol Lett 153:411–418

  14. Ishida K, Wang Y, Inoue Y (2001) Comonomer unit composition and thermal properties of poly(3-hydroxybutyrate-co-4-hydroxybutyrate) s biosynthesized by Ralstonia eutropha. Biomacromolecules 2:1285–1293

  15. Jo JE, Raj SM, Rathnasingh C, Selvakumar E, Jung WC, Park S (2008) Cloning, expression, and characterization of an aldehyde dehydrogenase from Escherichia coli K-12 that utilizes 3-hydroxypropionaldehyde as a substrate. Appl Microbiol Biotechnol 81:51–60

  16. Kato M, Bao HJ, Kang CK, Fukui T, Doi Y (1996) Production of a novel copolyester of 3-hydroxybutyric acid and medium chain length 3-hydroxyalkanaic acids by Pseudomonas sp 61–3 from sugars. Appl Microbiol Biotechnol 45:363–370

  17. Kim JS, Lee BH, Kim BS (2005) Production of poly(3-hydroxybutyrate-co-4-hydroxybutyrate) by Ralstonia eutropha. Biochem Eng J 23:169–174

  18. Kimura H, Ohura T, Takeishi M, Nakamura S, Doi Y (1999) Effective microbial production of poly(4-hydroxybutyrate) homopolymer by Ralstonia eutropha H16. Polym Int 48:1073–1079

  19. Kovach ME, Elzer PH, Hill DS, Robertson GT, Farris MA, Roop RM, Peterson KM (1995) Four new derivatives of the broad-host-range cloning vector pBBR1MCS, carrying different antibiotic-resistance cassettes. Gene 166:175–176

  20. Lee SY (1997) E. coli moves into the plastic age. Nat Biotechnol 15:17–18

  21. Lee SH, Oh DH, Ahn WS, Lee Y, Choi JI, Lee SY (2000) Production of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) by high-cell-density cultivation of Aeromonas hydrophila. Biotechnol Bioeng 67:240–244

  22. Lee WH, Azizan MNM, Sudesh K (2004) Effects of culture conditions on the composition of poly(3-hydroxybutyrate-co-4-hydroxybutyrate) synthesized by Comamonas acidovorans. Polym Degrad Stab 84:129–134

  23. Madison LL, Huisman GW (1999) Metabolic engineering of poly(3-hydroxyalkanoates): from DNA to plastic. Microbiol Mol Biol Rev 63:21–53

  24. Malaoui H, Marczak R (2000) Purification and characterization of the 1–3-propanediol dehydrogenase of Clostridium butyricum E5. Enzym Microb Technol 27:399–405

  25. Martin DP, Williams SF (2003) Medical applications of poly-4-hydroxybutyrate: a strong flexible absorbable biomaterial. Biochem Eng J 16:97–105

  26. Ouyang SP, Sun SY, Liu Q, Chen J, Chen GQ (2007) Microbial transformation of benzene to cis-3, 5-cyclohexadien-1, 2-diols by recombinant bacteria harboring toluene dioxygenase gene tod. Appl Microbiol Biotechnol 74:43–49

  27. Qiu YZ, Ouyang SP, Wu Q, Chen GQ (2004) Metabolic engineering for production of copolyesters consisting of 3-hydroxybutyrate and 3-hydroxyhexanoate by Aeromonas hydrophila. Macromol Biosci 4:255–261

  28. Qu XH, Wu Q, Zhang KY, Chen GQ (2006) In vivo studies of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) based polymers: biodegradation and tissue reactions. Biomaterials 27:3540–3548

  29. 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 43:1440–1446

  30. Saito Y, Nakamura S, Hiramitsu M, Doi Y (1996) Microbial synthesis and properties of poly(3-hydroxybutyrate-co-4-hydroxybutyrate). Polym Int 39:169–174

  31. Simon R, Priefer U, Pühler A (1983) A broad host range mobilization system for in vivo genetic engineering: transposon mutagenesis in gram negative bacteria. Nat Biotechnol 1:784–791

  32. Söhling B, Gottschalk G (1996) Molecular analysis of the anaerobic succinate degradation pathway in Clostridium kluyveri. J Bacteriol 178:871–880

  33. Song SS, Hein S, Steinbüchel A (1999) Production of poly(4-hydroxybutyric acid) by fed-batch cultures of recombinant strains of Escherichia coli. Biotechnol Lett 21:193–197

  34. Steinbüchel A, Valentin HE (1995) Diversity of bacterial polyhydroxyalkanoic acids. FEMS Microbiol Lett 128:219–228

  35. Sudesh K, Fukui T, Taguchi K, Iwata T, Doi Y (1999) Improved production of poly(4-hydroxybutyrate) by Comamonas acidovorans and its freeze-fracture morphology. Int J Biol Macromol 25:79–85

  36. Trendelenburg G, Strohle A (2005) Gamma-hydroxybutyrrate—a neurotransmitter, medicine, and drug. Nervenarzt 76:832–838

  37. Valentin HE, Reiser S, Gruys KJ (2000) Poly(3-hydroxybutyrate-co-4-hydroxybutyrate) formation from gamma-aminobutyrate and glutamate. Biotechnol Bioeng 67:291–299

  38. Wang YW, Mo WK, Yao HL, Wu Q, Chen GQ (2004) Biodegradation studies of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate). Polym Degrad Stab 85:815–821

  39. Weinel C, Nelson KE, Tummler B (2002) Global features of the Pseudomonas putida KT2440 genome sequence. Environ Microbiol 4:809–818

  40. Zaia DAM, Zaia C, Lichtig J (1998) Determination of total protein by spectrophotometry: advantages and disadvantages of proposed methods. Quim Nova 21:787–793

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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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Correspondence to Qiong Wu or Guo-Qiang Chen.

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Zhang, L., Shi, Z., 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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Keywords

  • 1,3-Propanediol dehydrogenase
  • 4-Hydroxybutyrate
  • PHB
  • P(4HB)
  • Poly-4-hydroxybutyrate
  • Poly(3-hydroxybutyrate-co-4-hydroxybutyrate)