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Towards repurposing the yeast peroxisome for compartmentalizing heterologous metabolic pathways

Author

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  • William C. DeLoache

    (UC Berkeley and UCSF Graduate Program in Bioengineering, University of California, Berkeley (UC Berkeley), Berkeley
    UC Berkeley)

  • Zachary N. Russ

    (UC Berkeley and UCSF Graduate Program in Bioengineering, University of California, Berkeley (UC Berkeley), Berkeley
    UC Berkeley)

  • John E. Dueber

    (UC Berkeley)

Abstract

Compartmentalization of enzymes into organelles is a promising strategy for limiting metabolic crosstalk and improving pathway efficiency, but improved tools and design rules are needed to make this strategy available to more engineered pathways. Here we focus on the Saccharomyces cerevisiae peroxisome and develop a sensitive high-throughput assay for peroxisomal cargo import. We identify an enhanced peroxisomal targeting signal type 1 (PTS1) for rapidly sequestering non-native cargo proteins. Additionally, we perform the first systematic in vivo measurements of nonspecific metabolite permeability across the peroxisomal membrane using a polymer exclusion assay. Finally, we apply these new insights to compartmentalize a two-enzyme pathway in the peroxisome and characterize the expression regimes where compartmentalization leads to improved product titre. This work builds a foundation for using the peroxisome as a synthetic organelle, highlighting both promise and future challenges on the way to realizing this goal.

Suggested Citation

  • William C. DeLoache & Zachary N. Russ & John E. Dueber, 2016. "Towards repurposing the yeast peroxisome for compartmentalizing heterologous metabolic pathways," Nature Communications, Nature, vol. 7(1), pages 1-11, September.
    • Handle: RePEc:nat:natcom:v:7:y:2016:i:1:d:10.1038_ncomms11152
    DOI: 10.1038/ncomms11152
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    Cited by:

    1. Briardo Llorente & Thomas C. Williams & Hugh D. Goold & Isak S. Pretorius & Ian T. Paulsen, 2022. "Harnessing bioengineered microbes as a versatile platform for space nutrition," Nature Communications, Nature, vol. 13(1), pages 1-7, December.
    2. Charlotte Cautereels & Jolien Smets & Peter Bircham & Dries De Ruysscher & Anna Zimmermann & Peter De Rijk & Jan Steensels & Anton Gorkovskiy & Joleen Masschelein & Kevin J. Verstrepen, 2024. "Combinatorial optimization of gene expression through recombinase-mediated promoter and terminator shuffling in yeast," Nature Communications, Nature, vol. 15(1), pages 1-17, December.

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