Orthologous promoters from related methylotrophic yeasts surpass expression of endogenous promoters of Pichia pastoris

Methylotrophic yeasts such as Komagataella phaffii (syn. Pichia pastoris , Pp ), Hansenula polymorpha ( Hp ), Candida boidinii ( Cb ) and Pichia methanolica ( Pm ) are widely used protein production platforms. Typically, strong, tightly regulated promoters of genes coding for their methanol utilizat...

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Bibliographic Details
Published in:AMB Express 2020-02, Vol.10 (1), p.38-38, Article 38
Main Authors: Vogl, Thomas, Fischer, Jasmin Elgin, Hyden, Patrick, Wasmayer, Richard, Sturmberger, Lukas, Glieder, Anton
Format: Article
Language:English
Subjects:
Alcohol oxidase
Biomedical and Life Sciences
Biotechnology
Depletion
Derepression
Gene expression
Komagataella phaffii
Life Sciences
Methanol
Methanol-free
Microbial Genetics and Genomics
Microbiology
Open reading frames
Original
Original Article
Orthologous promoters
Pichia pastoris
Promoters
Yeast
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Summary:Methylotrophic yeasts such as Komagataella phaffii (syn. Pichia pastoris , Pp ), Hansenula polymorpha ( Hp ), Candida boidinii ( Cb ) and Pichia methanolica ( Pm ) are widely used protein production platforms. Typically, strong, tightly regulated promoters of genes coding for their methanol utilization (MUT) pathways are used to drive heterologous gene expression. Despite highly similar open reading frames in the MUT pathways of the four yeasts, the regulation of the respective promoters varies strongly between species. While most endogenous Pp MUT promoters remain tightly repressed after depletion of a repressing carbon, Hp , Cb and Pm MUT promoters are derepressed to up to 70% of methanol induced levels, enabling methanol free production processes in their respective host background. Here, we have tested a series of orthologous promoters from Hp , Cb and Pm in Pp . Unexpectedly, when induced with methanol, the promoter of the HpMOX gene reached very similar expression levels as the strong methanol, inducible, and most frequently used promoter of the Pp alcohol oxidase 1 gene ( P PpAOX1 ). The HpFMD promoter even surpassed P PpAOX1 up to three-fold, when induced with methanol, and reached under methanol-free/derepressed conditions similar expression as the methanol induced P PpAOX1 . These results demonstrate that orthologous promoters from related yeast species can give access to otherwise unobtainable regulatory profiles and may even considerably surpass endogenous promoters in P. pastoris .
ISSN:2191-0855
2191-0855
DOI:10.1186/s13568-020-00972-1