Methanol Independent Expression by Pichia Pastoris Employing De-repression Technologies

This method for the first time describes reliable experimental procedures for efficient inducible methanol-free recombinant protein production in Pichia pastoris (Komagataella phaffii), employing the carbon source regulated promoters P_DCand P_DF in small scale experiments while cultivation parameters can be monitored online, and a constant glycerol feed is applied.