Inducible Synthetic Growth Regulation Using the ClpXP Proteasome Enhances cis,cis-Muconic Acid and Glycolic Acid Yields in Saccharomyces cerevisiae

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Journal Title
Journal ISSN
Volume Title
A1 Alkuperäisartikkeli tieteellisessä aikakauslehdessä
Date
2023-04-21
Major/Subject
Mcode
Degree programme
Language
en
Pages
1021–1033
Series
ACS Synthetic Biology, Volume 12, issue 4
Abstract
Engineered microbial cells can produce sustainable chemistry, but the production competes for resources with growth. Inducible synthetic control over the resource use would enable fast accumulation of sufficient biomass and then divert the resources to production. We developed inducible synthetic resource-use control overSaccharomyces cerevisiae by expressing a bacterial ClpXP proteasome from an inducible promoter. By individually targeting growth-essential metabolic enzymes Aro1, Hom3, and Acc1 to the ClpXP proteasome, cell growth could be efficiently repressed during cultivation. The ClpXP proteasome was specific to the target proteins, and there was no reduction in the targets when ClpXP was not induced. The inducible growth repression improved product yields from glucose (cis,cis-muconic acid) and per biomass (cis,cis-muconic acid and glycolic acid). The inducible ClpXP proteasome tackles uncertainties in strain optimization by enabling model-guided repression of competing, growth-essential, and metabolicenzymes. Most importantly, it allows improving production without compromising biomass accumulation when uninduced; therefore, it is expected to mitigate strain stability and low productivity challenges.
Description
Funding Information: Esa Tanhola and Anna-Liisa Ruskeepää are acknowledged for expert technical assistance in mass spectrometry analytics. Dr Tomi Määttä is acknowledged for valuable comments on proteomics data analysis. Dr Jussi Jäntti is acknowledged for valuable comments on strain engineering. The Proteomics Core Facility at EMBL Heidelberg is acknowledged and especially Mandy Rettel and Frank Stein for their expert support. Paula Jouhten acknowledges funding from the Academy of Finland (decision numbers 310514, 314125, 335783, 352410, and 352412). Natalia Kakko and Merja Penttilä acknowledge funding from the Jenny and Antti Wihuri Foundation (for the Center for Young Synbio Scientists). Publisher Copyright: © 2023 The Authors. Published by American Chemical Society.
Keywords
cis,cis-muconic acid, ClpXP proteasome, glycolic acid, Saccharomyces cerevisiae, synthetic regulation
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Citation
Kakko , N , Rantasalo , A , Koponen , T , Vidgren , V , Kannisto , M , Maiorova , N , Nygren , H , Mojzita , D , Penttilä , M & Jouhten , P 2023 , ' Inducible Synthetic Growth Regulation Using the ClpXP Proteasome Enhances cis,cis-Muconic Acid and Glycolic Acid Yields in Saccharomyces cerevisiae ' , ACS Synthetic Biology , vol. 12 , no. 4 , pp. 1021–1033 . https://doi.org/10.1021/acssynbio.2c00467