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

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Title
High-level production of recombinant human IFN-α2a with co-expression of tRNAarg(AGG/AGA) in high-cell-density cultures of Escherichia coli
Author(s)
Chul Soo Shin; Min Seon Hong; Hang Cheol Shin; Jee Won Lee
Bibliographic Citation
Biotechnology and Bioprocess Engineering, vol. 6, no. 4, pp. 301-305
Publication Year
2001
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.
Keyword
codon usageco-expression of argU geneescherichia colihigh-cell-density cultureshuman interferon-α2aplasmid instability
ISSN
1226-8372
Publisher
Springer
DOI
http://dx.doi.org/10.1007/BF02931994
Type
Article
Appears in Collections:
1. Journal Articles > Journal Articles
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