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Recombinant Streptomyces Lividans Secretes a Fusion Protein of Tendamistat and Proinsulin

Bio/Technology volume 7, pages 1055–1059 (1989)Cite this article

Abstract

We expressed a secreted protein fusion containing the complete α-amylase inhibitor of Streptomyces tendae (tendamistat, HOE 467) and proinsulin from the monkey, Macaca fascicularis, in Streptomyces lividans. The proinsulin gene was fused downstream of the tendamistat gene and separated by a linker sequence. The linker codes for a spacer peptide designed to allow independent folding of the two protein domains. After fermentation of recombinant strains, the fusion protein in the culture fluid had the expected molecular weight and reacted with antibodies against insulin and the inhibitor, respectively. The fusion protein still exhibits α-amylase inhibiting activity allowing for an easy selection of producing strains on agar plates. Additionally, biologically active des-Thr(B30) insulin was obtained after proteolytic digestion of the fusion protein with trypsin.

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Author notes

  1. Klaus-Peter Koller: Corresponding author.

Authors and Affiliations

  1. Hoechst AG, 6230, Frankfurt/M., 80, West Germany

    Klaus-Peter Koller, Günther Rieß, Klaus Sauber, Eugen Uhlmann & Holger Wallmeier

Authors
  1. Klaus-Peter Koller
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  2. Günther Rieß
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  3. Klaus Sauber
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  4. Eugen Uhlmann
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  5. Holger Wallmeier
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Koller, KP., Rieß, G., Sauber, K. et al. Recombinant Streptomyces Lividans Secretes a Fusion Protein of Tendamistat and Proinsulin. Nat Biotechnol 7, 1055–1059 (1989). https://doi.org/10.1038/nbt1089-1055

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