High-level production of recombinant human IFN-α2a with co-expression of tRNAArg(AGG/AGA) in high-cell-density cultures of Escherichia coli

Chul Soo Shin, Min Seon Hong, Hang Cheol Shin, Jeewon Lee

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

The co-expression of the argU gene in a double-vector expression system of recombinant Escherichia coli BL21(DE3)[pET-IFN2a+pAC-argU] significantly enhanced the production level of recombinant human interferon-α2a (rhIFN-α2a) in high cell density cultures, compared to a recombinant E. coli culture containing only the single expression vector, pET-IFN2a. The dry cell mass concentration increased to almost 100 g/L, and more than 4 g/L of rhIFN-α2a was accumulated in the culture broth. Evidently, the synthesis of rhIFN-α2a was strongly dependent on the pre-induction growth rate and more efficient at a higher specific growth rate. The additional supply of tRNAArg(AGG/AGA) enhanced the expression level of the rhIFN-α2a gene in the early stage of the post-induction phase, yet thereafter the specific production rate of rhIFN-α2a rapidly decreased due to severe segregational instability of plasmid vector pET-IFN2a. It would appear that the plasmid instability, which only occurred to pET-IFN2a in the double vector system, was related to the effect of translational stress due to the overexpression of rhIFN-α2a.

Original languageEnglish
Pages (from-to)301-305
Number of pages5
JournalBiotechnology and Bioprocess Engineering
Volume6
Issue number4
Publication statusPublished - 2001
Externally publishedYes

Fingerprint

RNA, Transfer, Arg
Interferons
Escherichia coli
Cell Culture Techniques
Cell Count
Plasmids
Genes
Growth
Gene Expression

Keywords

  • Co-expression of argU gene
  • Codon usage
  • Escherichia coli
  • High-cell-density cultures
  • Human interferon-α2a
  • Plasmid instability

ASJC Scopus subject areas

  • Biotechnology
  • Applied Microbiology and Biotechnology
  • Biomedical Engineering

Cite this

High-level production of recombinant human IFN-α2a with co-expression of tRNAArg(AGG/AGA) in high-cell-density cultures of Escherichia coli. / Shin, Chul Soo; Hong, Min Seon; Shin, Hang Cheol; Lee, Jeewon.

In: Biotechnology and Bioprocess Engineering, Vol. 6, No. 4, 2001, p. 301-305.

Research output: Contribution to journalArticle

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