Research Article | Volume: 6, Issue: 1, Jan-Feb, 2018

Nonconventional induction strategies for production of recombinant human tumor necrosis factor-alpha in Escherichia coli

Tapan Kumar Singha,  Pooja Gulati,  Sanjay Kumar   
Tapan Kumar Singha, Pooja Gulati, Sanjay Kumar

Department of Microbiology, Maharshi Dayanand University, Rohtak, Haryana, India.

Open Access   

Published:  Jan 17, 2018

DOI: 10.7324/JABB.2018.60105
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Abstract

The present study investigates the production kinetics of recombinant human tumor necrosis factor-alpha (TNF-α) in Escherichia coli under three different nonconventional inducible expression systems, namely, salt, the thermal, and auto-inducible system in batch culture in a bioreactor. A codon-optimized synthetic gene of human TNF-α was inserted into plasmid pET-14b under T7 promoter to obtain its plasmid-based expression in E. coli. Very high specific yield (~47% of total cellular protein (TCP)) was obtained with auto-inducible expression system. The thermal and salt-induced expressions could yield up to ~30% and ~17% of TCP, respectively. Maximum specific productivities were reported to be 0.092 g/gDCW/h, 0.1 g/gDCW/h, and 0.079 g/gDCW/h for auto-inducible, thermal-inducible, and salt-inducible expression systems, respectively.


Keyword:     Tumor necrosis factor-alpha Expression kinetics Auto-induction Thermal induction Salt induction Escherichia coli.

Citation:

Singha TK, Gulati P, Kumar S. Nonconventional induction strategies for production of recombinant human tumor necrosis factor-alpha in Escherichia coli. J App Biol Biotech. 2018;6(1):23-27. DOI: 10.7324/JABB.2018.60105

Copyright: Author(s). This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike license.
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Reference

  1. Dubravec DB, Spriggs DR, Mannick JA, Rodrick ML. Circulating human peripheral blood granulocytes synthesize and secrete tumor necrosis factor alpha. Proceedings of the National Academy of Sciences. 1990; 87(17):6758-6761. https://doi.org/10.1073/pnas.87.17.6758
  2. Carswell EA, Old LJ, Kassel RL, Green S, Fiore N, Williamson B. An endotoxin-induced serum factor that causes necrosis of tumors. Proceedings of the National Academy of Sciences. 1975; 72(9):3666-3670. https://doi.org/10.1073/pnas.72.9.3666
  3. Creasey, AA, Reynolds MT, Laird W. Cures and partial regression of murine and human tumors by recombinant human tumor necrosis factor. Cancer Research. 1986; 46 (11): 5687-5690.
  4. Ghezzi P, Cerami A. Tumor Necrosis Factor as a Pharmacological Target. Molecular Biotechnology. 2005; 31(3):239-244. https://doi.org/10.1385/MB:31:3:239
  5. Beutler B, Cerami A. Cachectin (Tumor Necrosis Factor): A Macrophage Hormone Governing Cellular Metabolism and Inflammatory Response. Endocrine Reviews. 1988; 9(1):57-66. https://doi.org/10.1210/edrv-9-1-57
  6. Moss ML, Sklair-Tavron L, Nudelman R. Drug Insight: tumor necrosis factor-converting enzyme as a pharmaceutical target for rheumatoid arthritis. Nature Clinical Practice Rheumatology. 2008; 4(6):300-309. https://doi.org/10.1038/ncprheum0797
  7. Chang J, Girgis L. Clinical use of anti-TNF-alpha biological agents--a guide for GPs. Australian Family Physician. 2007; 36 (12): 1035-1038.
  8. Reinhart K, Karzai W. Anti-tumor necrosis factor therapy in sepsis: Update on clinical trials and lessons learned. Critical Care Medicine. 2001; 29:S121-S125. https://doi.org/10.1097/00003246-200107001-00037
  9. Davis JM, Narachi MA, Alton NK, Arakawa T. Structure of human tumor necrosis factor alpha derived from recombinant DNA. Biochemistry. 1987; 26 (5):1322-1326. https://doi.org/10.1021/bi00379a018
  10. Marmenout A, Fransen L, Tavernier J, et al. Molecular cloning and expression of human tumor necrosis factor and comparison with mouse tumor necrosis factor. European Journal of Biochemistry. 1985; 152(3):515-522. https://doi.org/10.1111/j.1432-1033.1985.tb09226.x
  11. Wang D, Shi L. High-Level Expression, Purification, and In Vitro Refolding of Soluble Tumor Necrosis Factor-Related Apoptosis-Inducing Ligand (TRAIL). Applied Biochemistry and Biotechnology. 2008; 157(1):1-9. https://doi.org/10.1007/s12010-007-8079-x
  12. Lu, Z, Wang Z, Ming Y, Bao H, Liu X. High-level expression of the human tumor necrosis factor-alpha. Chinese Journal of Biotechnology. 1995; 11 (1): 37-44.
  13. Yamada M, Furutani Y, Notake M, Yamagishi UI, Yamayoshi M, Fukui T, Nomura H. et al. Efficient production of human tumour necrosis factor in Escherichia coli. Journal of Biotechnology. 1985; 3(3):141-153. https://doi.org/10.1016/0168-1656(85)90015-X
  14. Dobrynin VN, Berkova NP, Boldyreva EF, Bystrov NS, Kravchenko VV. Expression in Escherichia coli of DNA coding for human tumor necrosis factor. Bioorg Khim. 1988; 14(11): 1530-1537.
  15. Nakamura S, Masegi T, Kitai K, Ichikawa Y, Kudo T, Aono R, Horikoshi K. Extracellular production of human tumor necrosis factor-alpha by Escherichia coli using a chemically-synthesized gene. Agricultural and Biological Chemistry. 1990; 54(12):3241-3250. https://doi.org/10.1271/bbb1961.54.3241
  16. Lin Z, Lei H, Cao P. Expression, purification, and in vitro refolding of soluble tumor necrosis factor-related apoptosis-inducing ligand (TRAIL). Protein Expression and Purification. 2007; 51(2):276-282. https://doi.org/10.1016/j.pep.2006.07.026
  17. Han W, Zhang Y, Yan Z, Shi J. Construction of a new tumour necrosis factor fusion-protein expression vector for high-level expression of heterologous genes in Escherichia coli. Biotechnology and Applied Biochemistry. 2003; 37(2):109. https://doi.org/10.1042/BA20020070

  1. Menart V, Jevsevar S, Vilar M, Trobis A, Pavko A. Constitutive versus thermoinducible expression of heterologous proteins in Escherichia coli based on strong PR, PL promoters from phage lambda. Biotechnology and Bioengineering. 2003; 83(2):181-190. https://doi.org/10.1002/bit.10660
  2. Studier FW. Protein production by auto-induction in high-density shaking cultures. Protein Expression and Purification. 2005; 41(1):207-234. https://doi.org/10.1016/j.pep.2005.01.016
  3. Sambrook J, Green MR. Molecular Cloning: A Laboratory Manual; Cold Spring Harbor Laboratory Press, Cold Spring Harbour, New York. 2012, 157-210
  4. Tabors S, Richardson CC. A bacteriophage T7 RNA polymerase/promoter system for controlled exclusive expression of specific genes. Proceedings of the National Academy of Sciences. 1985; 82(4):1074-1078. https://doi.org/10.1073/pnas.82.4.1074
  5. Bhandari P, Gowrishankar J. An Escherichia coli host strain useful for efficient overproduction of cloned gene products with NaCl as the inducer. Journal of Bacteriology. 1997; 179 (13):4403-4406. https://doi.org/10.1128/jb.179.13.4403-4406.1997
  6. Zhang C, Liu Y, Zhao D, Li X, Yu R, and Su Z. Facile purification of Escherichia coli expressed tag-free recombinant human tumor necrosis factor alpha from supernatant. Protein Expression and Purification. 2014; 95: 195-203. https://doi.org/10.1016/j.pep.2013.12.012
  7. Grossman AD, Erickson JW, Gross CA. The htpR gene product of E. coli is a sigma factor for heat-shock promoters. Cell 1984; 38 (2):383-390. https://doi.org/10.1016/0092-8674(84)90493-8
  8. Weber J, Hoffmann F, Rinas U. Metabolic adaptation of Escherichia coli during temperature-induced recombinant protein production: 2. Redirection of metabolic fluxes. Biotechnology and Bioengineering 2002; 80(3):320-330. https://doi.org/10.1002/bit.10380
  9. Singha TK, Gulati P, Mohanty A, Khasa YP, Kapoor RK, and Kumar S. Efficient genetic approaches for improvement of plasmid based expression of recombinant protein in Escherichia coli: A review. Process Biochemistry. 2017; 55: 17-31. https://doi.org/10.1016/j.procbio.2017.01.026

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