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• W3047862007 abstract "The methylotrophic yeast Komagataella phaffii is among the most popular hosts for recombinant protein synthesis. Most recombinant proteins have been expressed in the wild-type Mut+ host strain from the methanol-inducible alcohol oxidase (AOX) promoter PAOX1. Since methanol metabolism has undesirable consequences, two additional host strains, Muts (Δaox1) and Mut- (Δaox1Δaox2), were introduced which consume less methanol and reportedly also express recombinant protein better than Mut+. Both results follow from a simple model based on two widespread assumptions, namely methanol is transported by diffusion and the sole inducer of PAOX1. To test this model, we studied 14C-methanol uptake in the Mut- strain and β-galactosidase expression in all three strains. We confirmed that methanol is transported by diffusion, but in contrast to the literature, Mut+ expressed β-galactosidase 5- and 10-fold faster than Muts and Mut-. These results imply that methanol is not the sole inducer of PAOX1-metabolites downstream of methanol also induce PAOX1. We find that formate or/and formaldehyde are probably true inducers since both induce PAOX1 expression in Mut- which cannot synthesize intracellular methanol from formate or formaldehyde. Formate offers a promising substitute for methanol since it does not appear to suffer from the deficiencies that afflict methanol. KEY POINTS: • This is the first study to systematically compare all three Mut phenotypes as host strains. • Mut+ strain expresses 5- and 10-fold faster than Muts and Mut- strains. • Methanol is transported by diffusion in Komagataella phaffii. • Formate and formaldehyde are true and strong inducers of PAOX1 expression." @default.
• W3047862007 created "2020-08-13" @default.
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• W3047862007 date "2020-08-06" @default.
• W3047862007 modified "2023-09-25" @default.
• W3047862007 title "The Mut+ strain of Komagataella phaffii (Pichia pastoris) expresses PAOX1 5 and 10 times faster than Muts and Mut− strains: evidence that formaldehyde or/and formate are true inducers of PAOX1" @default.
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• W3047862007 doi "https://doi.org/10.1007/s00253-020-10793-8" @default.
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