Expression and One-Step Purification of Recombinant Proteins Using an Alternative Episomal Vector for the Expression of N-Tagged Heterologous Proteins in Pichia pastoris

Abstract

Here we report the construction of an alternative episomal vector, pBGP3, which allows the expression of heterologous proteins with N-terminal hexahistidine and myc-epitope tags in Pichia pastoris. To test the usefulness of pBGP3, four cellulases from termites were expressed. Production was confirmed by activity assays and Western blot using anti-c-Myc antibody. Purification was performed by single-step Ni²⁺-affinity chromatography, which confirmed the efficiency of pBGP3.