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Expression of a β-Lactamase Preproinsulin Fusion Protein in Escherichia Coli

Bio/Technology volume 2, pages 165–168 (1984)Cite this article

Abstract

The level of expression of a β-lactamase human preproinsulin fusion protein1 in Escherichia coli was increased by plasmid and host manipulations. The initial expression level (0.01% of total protein) was increased two- to three-fold by replacing the p-lactamase promoter with two strong promoters: trp from Serratia marcescens and lacUV5 from E. coli. Replacement of the β-lactamase ribosome binding site (RBS) with a trp RBS enhanced the yield to a total of 0.16% of the total cell protein. Proinsulin was unstable, with a half-life of 14 minutes at 37°C. Proinsulin expression was increased three-fold (to about 0.5% of the total cell protein) by incubating the host at 30°C rather than 37°C.

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Authors and Affiliations

  1. Cetus Corporation, 1400 Fifty-Third Street, Emeryville, CA, 94608

    Anne W. Emerick, Barbara L. Bertolani, A. Ben-Bassat, Thomas J. White & Michael W. Konrad

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  1. Anne W. Emerick
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  2. Barbara L. Bertolani
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  3. A. Ben-Bassat
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  4. Thomas J. White
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  5. Michael W. Konrad
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Emerick, A., Bertolani, B., Ben-Bassat, A. et al. Expression of a β-Lactamase Preproinsulin Fusion Protein in Escherichia Coli. Nat Biotechnol 2, 165–168 (1984). https://doi.org/10.1038/nbt0284-165

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