Use of a mixture of glucose and methanol as substrates for the production of recombinant trypsinogen in continuous cultures with Pichia pastoris Mut+

► Glucose is a feasible partial substitute for methanol in recombinant protein production with Pichia pastoris Mut+ strains. ► The repressive effect of glucose on methanol utilization was eliminated in carbon/energy limited continuous cultures. ► With mixtures of glucose and methanol both yield of biomass per consumed substrate and productivity were enhanced. ► Production of porcine trypsinogen was only stable at low dilution rates.