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High level expression of interleukin-1 beta in a recombinant Escherichia coli strain for use in a controlled
P M Arthur1, B Duckworth, M Seidman
1Otsuka Pharmaceutical Co., Ltd., Rockville, Maryland 20850.
Abstract:
A recombinant bacterial strain for the large scale production of human interleukin-1 beta (IL-1 beta) was constructed. The lambda Pr and the tryptophan systems were compared for efficiency of transcription and regulation. The efficiency of IL-1 protein production from these constructs was analyzed. Enhanced protein synthesis was achieved by the fusion of lambda Pr promoter sequences with trp leader sequences which included the trp RBS. A strain (JM101) was selected for use as a host and tested in a one liter bioreactor. A growth and induction regimen was established for use in bioreactors which results in the accumulation of 0.75-0.95 g l-1 of recombinant IL-1.