The starch-binding domain as a tool for recombinant protein purification
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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.
KeywordsStarch-binding domain Purification tag Affinity tag Fusion protein Protein immobilization
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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