title: The starch-binding domain as a tool for recombinant protein purification.
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.
authors.affiliation: Departamento de Biología Molecular y Biotecnología, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, México, Distrito Federal 04510, México.
authors.forename: D
authors.initials: D
authors.lastname: Guillén
authors.forename: S
authors.initials: S
authors.lastname: Moreno-Mendieta
authors.forename: P
authors.initials: P
authors.lastname: Aguilera
authors.forename: S
authors.initials: S
authors.lastname: Sánchez
authors.forename: A
authors.initials: A
authors.lastname: Farres
authors.forename: R
authors.initials: R
authors.lastname: Rodríguez-Sanoja
mesh_terms: D010940:Plant Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics; D011485:Protein Binding; D011993:Recombinant Fusion Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D013213:Starch / Q000378:metabolism*
chemical_list: D010940:Plant Proteins; D011993:Recombinant Fusion Proteins; D013213:Starch