Science China Chemistry

, Volume 59, Issue 11, pp 1390–1396 | Cite as

Enhanced production of poly-3-hydroxybutyrate by recombinant Escherichia coli containing NAD kinase and phbCAB operon



With the help of Tn5 transposon technique, gene yfjB encoding NAD kinase in Escherichia coli (E. coli) was inserted into chromosome of recombinant E. coli polyhydroxybutyrate (PHB) containing PHB synthesis operon integrated in the host genome. After successful transposition of an extra yfjB gene copy into genome, the selected recombinant named E. coli PHBTY4 showed stronger NAD kinase activity than that of E. coli PHB. Shake flask studies suggested that both cell dry weight and PHB accumulation were significantly increased in E. coli PHBTY4 compared with that of the control. E. coli PHBTY4 produced approximately 23 g/L PHB compared with its control which synthesized only 10 g/L PHB when grown under the same conditions in a 6 L fermentor after 32 h of cultivation. In addition, E. coli PHBTY4 maintained high genetic stability during the cultivation processes. These results revealed a practical method to construct genetically stable strains harboring extra NAD kinase gene to enhance NADP(H)-dependent bio-reactions.


PHB polyhydroxyalkanoates Tn5 transposon NAD kinase Escherichia coli 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Chen GQ. Chem Soc Rev, 2009, 38: 2434–2446CrossRefGoogle Scholar
  2. 2.
    Chen GQ, Patel MK. Chem Rev, 2012, 112: 2082–2099CrossRefGoogle Scholar
  3. 3.
    Cao Q, Zhang J, Liu H, Wu Q, Chen J, Chen GQ. Biomaterials, 2014, 35: 8273–8283CrossRefGoogle Scholar
  4. 4.
    Abraham GA, Gallardo A, San Roman J, Olivera ER, Jodra R, Garcia B, Minambres B, Garcia JL, Luengo JM. Biomacromolecules, 2001, 2: 562–567CrossRefGoogle Scholar
  5. 5.
    Shen R, Cai L, Meng D, Wu L, Guo K, Dong G, Liu L, Chen J, Wu Q, Chen G. Sci China Life Sci, 2014, 57: 4–10CrossRefGoogle Scholar
  6. 6.
    Tan D, Xue YS, Aibaidula G, Chen GQ. Bioresour Technol, 2011, 102: 8130–8136CrossRefGoogle Scholar
  7. 7.
    Aldor IS, Keasling JD. Curr Opin Biotechnol, 2003, 14: 475–483CrossRefGoogle Scholar
  8. 8.
    Andreessen B, Lange AB, Robenek H, Steinbuchel A. Appl Environ Microbiol, 2010, 76: 622–626CrossRefGoogle Scholar
  9. 9.
    Meng DC, Wang Y, Wu LP, Shen R, Chen JC, Wu Q, Chen GQ. Metab Eng, 2015, 29: 189–195CrossRefGoogle Scholar
  10. 10.
    Liang Q, Zhang H, Li S, Qi Q. Appl Microbiol Biotechnol, 2011, 89: 57–62CrossRefGoogle Scholar
  11. 11.
    Keasling JD. ACS Chem Biol, 2008, 3: 64–76CrossRefGoogle Scholar
  12. 12.
    Wang T, Ma X, Zhu H, Li A, Du G, Chen J. Appl Microbiol Biotechnol, 2012, 93: 1853–1863CrossRefGoogle Scholar
  13. 13.
    Friehs K. Adv Biochem Eng Biotechnol, 2004, 86: 47–82Google Scholar
  14. 14.
    Tyo KE, Ajikumar PK, Stephanopoulos G. Nat Biotechnol, 2009, 27: 760–765CrossRefGoogle Scholar
  15. 15.
    Lemuth K, Steuer K, Albermann C. Microb Cell Fact, 2011, 10: 29CrossRefGoogle Scholar
  16. 16.
    Yin J, Fu XZ, Wu Q, Chen JC, Chen GQ. Appl Microbiol Biotechnol, 2014, 98: 8987–8997CrossRefGoogle Scholar
  17. 17.
    Yin J, Wang H, Fu XZ, Gao X, Wu Q, Chen GQ. Appl Microbiol Biotechnol, 2015, 99: 5523–5534CrossRefGoogle Scholar
  18. 18.
    Haldimann A, Wanner BL. J Bacteriol, 2001, 183: 6384–6393CrossRefGoogle Scholar
  19. 19.
    Kuhlman TE, Cox EC. Nucleic Acids Res, 2010, 38: e92CrossRefGoogle Scholar
  20. 20.
    Reznikoff WS. Annu Rev Microbiol, 1993, 47: 945–963CrossRefGoogle Scholar
  21. 21.
    Dulermo R, Onodera T, Coste G, Passot F, Dutertre M, Porteron M, Confalonieri F, Sommer S, Pasternak C. PLoS One, 2015, 10: e0124358CrossRefGoogle Scholar
  22. 22.
    Xia Z, Zhang W, Lei L, Liu X, Wei HL. Appl Microbiol Biotechnol, 2015, 99: 6503–6514CrossRefGoogle Scholar
  23. 23.
    Yu BJ, Kim C. Methods Mol Biol, 2008, 416: 261–277CrossRefGoogle Scholar
  24. 24.
    Li ZJ, Cai L, Wu Q, Chen GQ. Appl Microbiol Biotechnol, 2009, 83: 939–947CrossRefGoogle Scholar
  25. 25.
    Fukui T, Yoshimoto A, Matsumoto M, Hosokawa S, Saito T. Arch Microbiol, 1976, 110: 149–156CrossRefGoogle Scholar
  26. 26.
    Reynen M, Reipen I, Sahm H, Sprenger GA. Mol Gen Genet, 1990, 223: 335–341CrossRefGoogle Scholar
  27. 27.
    Wu J, Xu J, Hong Q, Li S. Wei Sheng Wu Xue Bao, 2008, 48: 45–50Google Scholar
  28. 28.
    de Lorenzo V, Herrero M, Jakubzik U, Timmis KN. J Bacteriol, 1990, 172: 6568–6572CrossRefGoogle Scholar
  29. 29.
    Datsenko KA, Wanner BL. Proc Natl Acad Sci USA, 2000, 97: 6640–6645CrossRefGoogle Scholar
  30. 30.
    Yanisch-Perron C, Vieira J, Messing J. Gene, 1985, 33: 103–119CrossRefGoogle Scholar
  31. 31.
    McGuinness ET, Butler JR. Int J Biochem, 1985, 17: 1–11CrossRefGoogle Scholar
  32. 32.
    Kawai S, Mori S, Mukai T, Hashimoto W, Murata K. Eur J Biochem, 2001, 268: 4359–4365CrossRefGoogle Scholar
  33. 33.
    Shi F, Li K, Huan X, Wang X. Appl Biochem Biotechnol, 2013, 171: 504–521CrossRefGoogle Scholar
  34. 34.
    Higashitani A, Higashitani N, Yasuda S, Horiuchi K. Nucleic Acids Res, 1994, 22: 2426–2427CrossRefGoogle Scholar
  35. 35.
    Martin VJ, Mohn WW. J Microbiol Methods, 1999, 35: 163–166CrossRefGoogle Scholar
  36. 36.
    Grose JH, Joss L, Velick SF, Roth JR. Proc Natl Acad Sci USA, 2006, 103: 7601–7606CrossRefGoogle Scholar
  37. 37.
    Lim SJ, Jung YM, Shin HD, Lee YH. J Biosci Bioeng, 2002, 93: 543–549CrossRefGoogle Scholar
  38. 38.
    Simon R, Priefer U, Pühler A. Nat Biotechnol, 1983, 1: 784–791CrossRefGoogle Scholar
  39. 39.
    Cong YG, Xu QW. Acta Academiae Medicinae Militaris Tertiae, 2004, 26: 1145–1147Google Scholar

Copyright information

© Science China Press and Springer-Verlag Berlin Heidelberg 2016

Authors and Affiliations

  1. 1.Center for Synthetic and Systems Biology, School of Life Science, Tsinghua-Peking Center for Life SciencesTsinghua UniversityBeijingChina
  2. 2.Beijing Key Laboratory of Bioprocess, College of Life Science and TechnologyBeijing University of Chemical TechnologyBeijingChina
  3. 3.Center for Nano and Micro MechanicsTsinghua UniversityBeijingChina

Personalised recommendations