Production of a soluble and functional recombinant streptavidin in Escherichia coil

Anna Gallizia, Claudia De Lalla, Errica Nardone, Paolo Santambrogio, Anna Brandazza, Alessandro Sidoli, Paolo Arosio

Research output: Contribution to journalArticlepeer-review


The cDNA for streptavidin (residues 15-159) was subcloned into an expression vector in fusion at the N-terminus with the T7-tag (12 residues). Conditions were found to express the protein in Escherichia coli in a soluble, assembled, and active form. The protein was purified in two simple steps which involved heating at 75°C and affinity chromatography on iminobiotin agarose. The purified protein was obtained in yields of 70 mg per liter of bacterial culture. Electron spray mass spectrometry analysis showed that the recombinant streptavidin had the expected molecular mass without covalent modifications. ELISA and surface plasmon resonance analyses showed it to be functionally analogous to the natural streptavidin. This appears to be an improvement over the reported methods of recombinant streptavidin production which involve protein renaturation or the use of eukaryotic expression systems.

Original languageEnglish
Pages (from-to)192-196
Number of pages5
JournalProtein Expression and Purification
Issue number2
Publication statusPublished - Nov 1998


  • Avidin
  • Biotin
  • Recombinant protein
  • Streptavidin

ASJC Scopus subject areas

  • Biochemistry


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