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Applied Microbiology and Biotechnology

, Volume 97, Issue 9, pp 4141–4148 | Cite as

The starch-binding domain as a tool for recombinant protein purification

  • D. Guillén
  • S. Moreno-Mendieta
  • P. Aguilera
  • S. Sánchez
  • A. Farres
  • R. Rodríguez-SanojaEmail author
Methods and protocols

Abstract

Recombinant protein purification with affinity tags is a widely employed technique. One of the most common tags used for protein purification is the histidine tag (Histag). In this work, we use a tandem starch-binding domain (SBDtag) as a tag for protein purification. Four proteins from different sources were fused to the SBDtag, and the resulting fusion proteins were purified by affinity chromatography using the Histag or the SBDtag. The results showed that the SBDtag is superior to the Histag for protein purification. The efficient adsorption of the fusion proteins to raw corn starch was also demonstrated, and two fusions were selected to test purification directly using raw starch from rice, corn, potato, and barley. The two fusion proteins were successfully recovered from crude bacterial extract using raw starch, thus demonstrating that the SBDtag can be used as an efficient affinity tag for recombinant protein purification on an inexpensive matrix.

Keywords

Starch-binding domain Purification tag Affinity tag Fusion protein Protein immobilization 

Notes

Acknowledgment

D. Guillén, S Moreno-Mendieta, and P. Aguilera were supported by personal grants from Consejo Nacional de Ciencia y Tecnología (CONACyT) México. This work is supported by UNAM-DGAPA grants IN209410-3, IN222113, and CONACYT grant 131149. We thank Beatriz Ruiz, Laura Escalante, and María Elena Munguía for technical assistance.

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Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • D. Guillén
    • 1
    • 2
  • S. Moreno-Mendieta
    • 1
    • 3
  • P. Aguilera
    • 1
  • S. Sánchez
    • 1
  • A. Farres
    • 4
  • R. Rodríguez-Sanoja
    • 1
    Email author
  1. 1.Departamento de Biología Molecular y Biotecnología, Instituto de Investigaciones BiomédicasUniversidad Nacional Autónoma de MéxicoMéxicoMéxico
  2. 2.Programa de Posgrado en Ciencias BioquímicasUniversidad Nacional Autónoma de MéxicoMéxicoMéxico
  3. 3.Programa de Doctorado en Ciencias BiomédicasUniversidad Nacional Autónoma de MéxicoMéxicoMéxico
  4. 4.Departamento de Alimentos y Biotecnología, Facultad de QuímicaUniversidad Nacional Autónoma de MéxicoMéxicoMéxico

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