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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

Authors 

Chul Soo ShinMin Seon HongHang Cheol ShinJee Won Lee

Publisher 

Springer Verlag (Germany)

Issue Date 

2001

Citation 

Biotechnology and Bioprocess Engineering, vol. 6, no. 4, pp. 301-305

Keywords 

codon usageco-expression of argU geneescherichia colihigh-cell-density cultureshuman interferon-α2aplasmid instability

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.

ISSN 

1226-8372

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2017-04-19


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