Skip to main content

High-level production of soluble tumor necrosis factor-related apoptosis-inducing ligand (Apo2L/TRAIL) in high-density cultivation of recombinant Escherichia coli using a combined feeding strategy

Abstract

Recombinant Escherichia coli strain C600/pBV-TRAIL (encoding for 114-281 amino acids of TRAIL's soluble fragment) produced a recombinant human tumor necrosis factor-related apoptosis-inducing ligand (Apo2L/TRAIL). Using a combined strategy of exponential feeding and pH-stat feeding, high concentrations of biomass (65 g dry wt l−1) and active soluble TRAIL (1.4 g l−1) were obtained within 30 h. The accumulation of acetate, which usually occurs during the process of high-density culture of Escherichia coli and especially in the induction stage of protein synthesis, was avoided.

This is a preview of subscription content, access via your institution.

References

  • Castan A, Enfors SO (2000) Characteristics of a DO-controlled fedbatch culture of Escherichia coli. Bioproc. Eng. 22: 509–515.

    Google Scholar 

  • Laemmi UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685.

    Google Scholar 

  • Lee SY (1996) High cell-density culture of Escherichia coli. Trends Biotechnol. 14: 98–105.

    Google Scholar 

  • Pan JG, Rhee JS, Leheault JM (1987) Physiological constraints in increasing biomass concentration of Escherichia coli B in fedbatch culture. Biotechnol. Lett. 9: 89–94.

    Google Scholar 

  • Schmidt M, Babu KR, Khanna N, Marten S, Rinas U (1999) Temperature-induced production of recombinant human insulin in high-cell density cultures of recombinant Escherichia coli. J. Biotechnol. 68: 71–83.

    Google Scholar 

  • Seeger A, Schneppe B, McCarthy JEG, Deckwer WD, Rinas U (1995) Comparison of temperature and isopropyl-â-D-thiogalacto-pyranoside-induced synthesis of basic fibroblast growth factor in high-cell-density cultures of recombinant Escherichia coli. Enzyme Microb. Technol. 17: 947–953.

    Google Scholar 

  • Walczak H, Miller RE, Arial K, Gliniak B, Griffith TS, Kubin M, Chin W, Jones J, Woodward A, Le T, Smith C, Smolak P, Goodwin RG, Rauch CT, Schuh JCL, Lynch DH (1999) Tumoricidal activity of tumor necrosis factor-related apoptosis-inducing ligand in vivo. Nat. Med. 5: 157–163.

    Google Scholar 

  • Wiley SR, Schooley K, Smolak PJ, Din WS, Huang CP, Nicholl JK, Sutherland GR, Smith TD, Rauch C, Smith CA (1995) Identification and characterization of a new member of the TNF family that induces apoptosis. Immunity 3: 673–682.

    Google Scholar 

  • Yin L, Jian C, Ying YM, Shi YL, Yoon MK (1998) Effect of additives and fed-batch culture strategies on the production of glutathione by recombinant Escherichia coli. Proc. Biochem. 33: 709–714.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Shen, YL., Zhang, Y., Sun, AY. et al. High-level production of soluble tumor necrosis factor-related apoptosis-inducing ligand (Apo2L/TRAIL) in high-density cultivation of recombinant Escherichia coli using a combined feeding strategy. Biotechnology Letters 26, 981–984 (2004). https://doi.org/10.1023/B:BILE.0000030043.84226.ac

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/B:BILE.0000030043.84226.ac

  • combined feeding strategy
  • Escherichia coli
  • fed-batch
  • high-cell-density cultivation
  • TRAIL