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Molecular Biotechnology

, Volume 9, Issue 2, pp 99–106 | Cite as

Overproduction of soluble, extracellular cytotoxin α-sarcin inEscherichia coli

  • Dino Parente
  • Giuseppe Raucci
  • Laura D'Alatri
  • Guy d'Estais
  • Sabrina Novelli
  • Aurelio Pacilli
  • Maria Pia Saccinto
  • Antonio Mele
  • Rita De Santis
Research

Abstract

The goal of the present study was to establish the condition to obtain preparative amounts of the recombinant cytotoxin α-sarcin to be used for immunoconjugate production.

α-Sarcin cDNA was isolated fromAspergillus giganteus strain MDH 18894 and its expression inEscherichia coli was attempted by the use of both two-cistron and fusion protein-expression systems. Whereas the former resulted in low intracellular expression level of recombinant α-sarcin (r-Sar), the latter allowed high-level expression of the fusion protein in the culture supernant.

A variant form of α-sarcin with an additional threonine residue in position 1 (Thr-Sar) was obtained by proteolytic processing of the fusion protein with a final yield after purification of 40 mg/L of culture. Both recombinant proteins r-Sar and Thr-Sar were identical to native a-sarcin with respect to the biochemical properties and to the in vitro biological activity.

Index Entries

Ribosome-inactivating protein Aspergillus giganteus toxin E. coli expression recombinant protein secretion 

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Copyright information

© Humana Press Inc 1998

Authors and Affiliations

  • Dino Parente
    • 1
  • Giuseppe Raucci
    • 1
  • Laura D'Alatri
    • 1
  • Guy d'Estais
    • 2
  • Sabrina Novelli
    • 2
  • Aurelio Pacilli
    • 1
  • Maria Pia Saccinto
    • 1
  • Antonio Mele
    • 1
  • Rita De Santis
    • 1
  1. 1.Department of BiotechnologyMenarini Ricerche S.p.A.Pomezia (Rome)Italy
  2. 2.SUDBIOTEC S.c.r.l.Pomezia (Rome)Italy

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