Increased dosage of AOX1 promoter-regulated expression cassettes leads to transcription attenuation of the methanol metabolism in Pichia pastoris

The methanol-regulated alcohol oxidase promoter (P AOX1 ) of Pichia pastoris is one of the strongest promoters for heterologous gene expression in this methylotrophic yeast. Although increasing gene dosage is one of the most common strategies to increase recombinant protein productivities, the increase of gene dosage of Rhizopus oryzae lipase ( ROL ) in P. pastoris has been previously shown to reduce cell growth, lipase production and substrate...