Towards repurposing the yeast peroxisome for compartmentalizing heterologous metabolic pathways

DeLoache, William C.; Russ, Zachary N.; Dueber, John E.

doi:10.1038/ncomms11152

Title: Towards repurposing the yeast peroxisome for compartmentalizing heterologous metabolic pathways

Journal Article · Wed Mar 30 00:00:00 EDT 2016 · Nature Communications

DOI:https://doi.org/10.1038/ncomms11152· OSTI ID:1255249

DeLoache, William C. ^[1];

^[1]; Dueber, John E. ^[1]

Univ. of California, Berkeley, CA (United States)

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. Lastly, 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.

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Research Organization:: University of California, Berkeley, CA (United States)

Sponsoring Organization:: USDOE Office of Science (SC)

Grant/Contract Number:: SC0008084

OSTI ID:: 1255249

Journal Information:: Nature Communications, Vol. 7; ISSN 2041-1723

Publisher:: Nature Publishing GroupCopyright Statement

Country of Publication:: United States

Language:: English

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Cited by: 93 works

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