Expression and characterization of a thermophilic trehalose synthase from meiothermus ruber cbs-01 in Pichia pastoris

Abstract

Trehalose synthase (TreS) was proved to catalyze the reversible reaction of maltose into trehalose by intramolecular transglucosylation. In this work, a yeast expression system was constructed to express TreS from Meiothermus ruber CBS-01 in the eukaryotic Pichia pastoris expression system. The TreS gene with 6× His tag at the 3 end was subcloned into the eukaryotic expression vector pPIC3.5K. Then the constructed vector was integrated into Pichia pastoris strain KM71. The recombinant was induced by sterile methanol and the bioactive TreS was expressed successfully intracellular. After optimizing culture conditions, we got approximately 150 mg/L recombinant protein. It was the first time to express TreS from M. ruber CBS-01 in eukaryotic expression system. The purified TreS was also characterized in details. © Springer-Verlag Berlin Heidelberg 2014.