Large-scale production, purification, and characterisation of recombinant Phaseolus vulgaris phytohemagglutinin E-form expressed in the methylotrophic yeast Pichia pastoris

摘要: The kidney bean lectin Phaseolus vulgaris phytohemagglutinin E-form (PHA-E) was expressed and secreted by the methylotrophic yeast Pichia pastoris. To optimise yields of PHA-E, transformants P. pastoris were selected for high-level production recombinant protein. A scaleable process purification gram quantities PHA-E is reported. at approximately 100 mg/L 2- 200-L scale purified to 95% homogeneity in a single step using cation-exchange chromatography. consists four forms with molecular masses between 28.5 31.5 kDa, as assessed MALDI-TOF, whereas its native counterpart has mass 30.5 kDa. Endoglycosidase treatment revealed that range size protein attributed differences nature N-linked oligosaccharides bound primary amino acid sequence found be identical have an agglutination activity similar PHA-E. data presented here suggest that, pastoris, can produced synthesised plants respect structure biological activity.