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The Tendamistat Expression-Secretion System: Synthesis of Proinsulin Fusion Proteins with Streptomyces Lividans

  • Klaus-P. Koller
  • Günther Rieß
  • Klaus Sauber
  • László Vértesy
  • Eugen Uhlmann
  • Holger Wallmeier
Part of the Federation of European Microbiological Societies Symposium Series book series (FEMS, volume 55)

Abstract

Tendamistat (HOE 467), a potent inhibitor of the human pancreatic α-amylase, is an acidic protein of 74 amino acids (Vértesy et al., 1984). The inhibitor gene was cloned from an amplified genomic sequence of an over-producing strain of Streptomyces tendae and further characterized. By expression of the gene in the heterologous host Streptomyces lividans we demonstrated that secretion of this protein was mediated by a signal peptide dependent mechanism (Koller & Rieß, 1989). We have also evaluated the tendamistat-based secretion system for a number of foreign proteins, for example interleukin II and proinsulin (Bender et al., 1990, Koller et al., 1989). Our main interest focussed on the formation of disulphide bonds, stability and activity of secreted proteins.

Keywords

Disulphide Bridge Amylase Inhibitor Space Peptide Proinsulin Gene Secrete Fusion Protein 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Literature

  1. Aretz, W., Koller, K.-P., and Riess, G., 1989. Proteolytic enzymes from recombinant Streptomyces lividans TK24. 1989. FEMS Microbiology Letters 65: 31CrossRefGoogle Scholar
  2. Bender, E., Koller, K.-P., and Engels, J.W., 1990. Secretory synthesis of human interleukin-2 by Streptomyces lividans. Gene, 86: 227PubMedCrossRefGoogle Scholar
  3. Brawner, M., Taylor, D., and Fornwald, J. 1990. Expression of the soluble CD-4 receptor in Streptomyces. J. Cellular Biochemistry, Supplement 14A: 103Google Scholar
  4. Chang, S.-Y., and Chang, S. 1988. Secretion of heterologous proteins in Streptomyces lividans. Seventh Int. Symp. on Biology of Actinomycetes. Y. Okami, T. Beppu, H. Ogawara (Eds). Japan Scientific Societies Press, Tokyo. p. 103Google Scholar
  5. Koller, K.-P., Engels, J.W., and Uhlmann, E. 1984. Gene amplification and over-production of the a-amylase inhibitor (Hoe 467, Tendamistat) in Streptomyces tendae. Third European Congress on Biotechnology, Vol. III: 273Google Scholar
  6. Koller, K.-P., and Riess, G., 1989. Heterologous expression of the a-amylase inhibitor gene cloned from an amplified genomic sequence of Streptomyces tendae. J.Bacteriol. Vol. 171, No. 9: 4953PubMedGoogle Scholar
  7. Koller, K.-P., Riess, G., Sauber, K., Uhlmann, E., and Wallmeier, H., 1989. Recombinant Streptomyces lividans secretes a fusion protein of tendamistat and proinsulin. Bio/Technology Vol. 7: 1055Google Scholar
  8. Steiert, J.G., Pogell, B.M., Speedie, M.K., and Laredo, J. 1989. A gene coding for a membrane-bound hydrolase is expressed as a secreted, soluble enzyme in Streptomyces lividans. Bio/Technology, 7: 65CrossRefGoogle Scholar
  9. Thim, L., Hansen, M.T., Norris, K., Hoegh,I., Boel, E., Forstrom, J., Ammerer, G., and Fiil, N.P. 1986. Secretion and processing of insulin precursors in yeast. Proc. Natl. Acad. Sci. USA 83: 6766PubMedCrossRefGoogle Scholar
  10. Thim, L., Snel, L., Norris, K., and Hansen, M.T. 1989. Insulin and insulin analogs from Saccharomyces cerevisiae. In: Genetics and Molecular Biology of Industrial Microorganisms. C.L. Hershberger, S.W. Queener, G. Hegemann (Eds).American Scociety for Microbiology, Washington p. 322Google Scholar
  11. Vértesy,L., Oeding,V., Bender,R., Zepf,K. and Nesemann,G. 1984. Tendamistat (HOE 467), a tight binding a-amylase inhibitor from Streptomyces tendae 4158. Isolation and biochemical properties. Eur. J. Biochem. 141: 505PubMedCrossRefGoogle Scholar
  12. Vértesy,L., Tripier,D., Koller,K.-P. and Riess,G. 1991. Disulphide bridge formation of proinsulin fusion proteins during secretion in Streptomyces. Biol. Chem. Hoppe-Seyler 372: in pressGoogle Scholar

Copyright information

© Plenum Press, New York 1991

Authors and Affiliations

  • Klaus-P. Koller
    • 1
  • Günther Rieß
    • 1
  • Klaus Sauber
    • 1
  • László Vértesy
    • 1
  • Eugen Uhlmann
    • 1
  • Holger Wallmeier
    • 1
  1. 1.HOECHST AGFrankfurt/Main 80Germany

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