Biotechnology and Bioprocess Engineering:BBE

The Korean Society for Biotechnology and Bioengineering (한국생물공학회)
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High-level Production of Recombinant Human IFN-$\alpha2a$ with Co-expression of $tRNA^{Arg(AFF/AGA)}$ in High-cell-density Cultures of Escherichia coli

  • Shin, Chul-Soo (Bioprocess Engineering Laboratory, Hanhyo Institutes of Technology, Yusong, Taejon 305-390) ;
  • Hong, Min-Seon (Bioprocess Engineering Laboratory, Hanhyo Institutes of Technology, Yusong, Taejon 305-390) ;
  • Shin, Hang-Chel (Department of Life Science and Bioinformatics, Soongsil University, Seoul 156-743) ;
  • Lee, Jeewon (Laboratory of Microbial and Bioprocess Engineering, Korea Research Institute of Bioscience and Biotechnology(KRIBB). P. O. Box 115, Yusong, Taejon 305)
  • Published : 2001.07.01
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Abstract

The co-expression of the arg U gene in a double-vector expression system of recombi-nant Escherichia coli BL22(DE3)[pET-IEN2a+pAC-argU] significantly enhanced the production level of reconminant human interferon -$\alpha$2a(rhIFN-$\alpha$2a) in high cell density cultures, compared to a recombinant E. coli culture containing only the single expression vector, pET-IEN2a. The dry cell mass concentration increased to almost 100 g/L, and more than 4 g/L of rhIFN-$\alpha$2a was accumu-lated in the culture broth. Evidently, the synthesis of rhIFN-$\alpha$2a was strongly dependent on the pre-induction growtih rate and more efficient at a higher specific growth rate. The additional sup-ply of tRN $A^{Arg(AGG/AGA)}$ enhanced the expression level of the rhIFN-$\alpha$2a gene in the early stage of the post-induction phase, yet thereafter the specific production rate of rhIFN-$\alpha$2a rapidly de-creased due to severe segregational instability of plasmid vector pET-IEN2a. It would appear that the plasmid instability with only occurred to pET-IEN2a in the double vector system, was re-lated to the effect of translational stress due to the over expression of rhIFN-$\alpha$2a.

Keywords

References

  1. Trends Biotechnol. v.8 High expression of cloned genes in E. coli and its consequences. Carrier, M. J.;M. E. Nugent;W. C. A. Tacon;S. B. Primrose
  2. Enzyme Microb. Technol. v.8 Factors influencing productivity of fermentations applying recombinant microorganisms. Zabriskie, D. W.;E. J. Arcuri
  3. AIChE J. v.34 Optimizing the production of recombinant proteins in microorganisms. Georgiou, G.
  4. Gene v.85 Highlevel expression of recombinant genes in Escherichia coli is dependent on the availability of the dnaY gene product. Brinkmann U.;R. E. Matters;P. Buckel
  5. Biochem. Mol. Biol. Int. v.32 Enhancement of expression of human granulocytemacrophage colony stimulating factor by argU gene product in Escherichia coli. Hua, Z.;H. Wang;D. Chen;Y. Chen;D. Zhu
  6. Curr.Opin. Biotechnol. v.6 Effects of rare codon clusters on highlevel expression of heterologous proteins in Escherichia coli. Kane, J. F.
  7. Cell v.45 The E. coli dnaY gene encodes an arginine transfer RNA. Garcia, G. M.;P. K. Mar;D. A. Mullin;J. R. Walker;N. E. Prather
  8. Annu. Rev. Biochem. v.56 Interferons and their actions. Peska, S.;J. A. Langer;K. C. Zoon;C. E. Samuel
  9. Biotechnol. Lett. v.20 Supply of the argU gene product allows high-level expression of recombinant human interferon-α2a in Escherichia coli. Jeong, W.;H. C. Shin
  10. Biotechnol. Bioeng. v.69 Glucagon-induced self-association of recombinant proteins in Escherichia coli and affinity purification using a fragment of glucagon receptor. Kim, D-Y.;J. Lee;V. Saraswat;Y-H. Park
  11. FEMS Microbiol. Lett. v.179 Effect of specific production rate of recombinant protein on multimerization of plasmid vector and gene expression level. Saraswat, V.;D. Y. Kim;J. Lee;Y. H. Park
  12. Microbiol. Rev. v.60 Strategies for Achieving Highlevel expression of genes in Escherichia coli. Makrides, S. C.
  13. Appl. Microbiol. Biotechnol. v.49 Growth-associated synthesis of recombinant human glucagon and human growth hormone in high-celldensity cultures of Escherichia coli. Shin, C. S.;M. S. Hong;D. Y. Kim;H. C. Shin;J. Lee
  14. Symp. Soc. Gen. Mictobiol. v.11 Microbiological reactions to the environment. Herbert D.
  15. J. Gen. Microbiol. v.136 Effect of growth rate on plasmid maintenance by Escherichia coli HB101(pAT153). Brownline L.;J. R. Stephenson;J. A. Cole
  16. Plasmid v.24 Presence of the bacterial hemoglobin gene improves α-amylase production of a recombinant Escherichia coli strain. Khosravi M.;D. A. Webster;B. C. Stark
  17. J. Ind. Microbiol. Biotechnol. v.22 Synthesis of mini-proinsulin precursors using Ntermini of human TNF α as fusion partners in recombinant Escherichia coli. Shin, C. S.;M. S. Hong;D. Y. Kim;J. Lee;Y. H. Park