Efficient production of active chicken avidin using a bacterial signal peptide in Escherichia coli

VR Hytönen, OH Laitinen, Tomi Airenne, H Kidron, NJ Meltola, EJ Porkka, J Hörhä, T Paldanius, Määttä JAE, HR Nordlund, Mark S Johnson, Tiina Salminen, KJ Airenne, S Ylä-Herttuala, MS Kulomaa

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    Chicken avidin is a highly popular tool with countless applications in the life sciences. In the present study, an efficient method for producing avidin protein in the periplasmic space of Escherichia coli in the active form is described. Avidin was produced by replacing the native signal sequence of the protein with a bacterial OmpA secretion signal. The yield after a single 2-iminobiotin-agarose affinity purification step was approx. 10 mg/l of virtually pure avidin. Purified avidin had 3.7 free biotin-binding sites per tetramer and showed the same biotin-binding affinity and thermal stability as egg-white avidin. Avidin crystallized under various conditions, which will enable X-ray crystallographic studies. Avidin produced in E. coli lacks the carbohydrate chains of chicken avidin and the absence of glycosylation should decrease the nonspecific binding that avidin exhibits towards many materials [Rosebrough and Hartley (1996) J. Nucl. Med. 37, 1380-1384]. The present method provides a feasible and inexpensive alternative for the production of recombinant avidin, avidin mutants and avidin fusion proteins for novel avidin-biotin technology applications.
    Original languageUndefined/Unknown
    Pages (from-to)385–390
    Number of pages6
    JournalBiochemical Journal
    Issue number2
    Publication statusPublished - 2004
    MoE publication typeA1 Journal article-refereed


    • bacterial
    • chicken avidin
    • expression system
    • Biotin
    • avidin-biotin technology

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