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Pathway Engineering, Re-targeting, and Synthetic Scaffolding Improve the Production of Squalene in Plants

[Image: see text] Plants are increasingly becoming an option for sustainable bioproduction of chemicals and complex molecules like terpenoids. The triterpene squalene has a variety of biotechnological uses and is the precursor to a diverse array of triterpenoids, but we currently lack a sustainable...

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Autores principales: Bibik, Jacob D., Weraduwage, Sarathi M., Banerjee, Aparajita, Robertson, Ka’shawn, Espinoza-Corral, Roberto, Sharkey, Thomas D., Lundquist, Peter K., Hamberger, Björn R.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Chemical Society 2022
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9208017/
https://www.ncbi.nlm.nih.gov/pubmed/35549088
http://dx.doi.org/10.1021/acssynbio.2c00051
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author Bibik, Jacob D.
Weraduwage, Sarathi M.
Banerjee, Aparajita
Robertson, Ka’shawn
Espinoza-Corral, Roberto
Sharkey, Thomas D.
Lundquist, Peter K.
Hamberger, Björn R.
author_facet Bibik, Jacob D.
Weraduwage, Sarathi M.
Banerjee, Aparajita
Robertson, Ka’shawn
Espinoza-Corral, Roberto
Sharkey, Thomas D.
Lundquist, Peter K.
Hamberger, Björn R.
author_sort Bibik, Jacob D.
collection PubMed
description [Image: see text] Plants are increasingly becoming an option for sustainable bioproduction of chemicals and complex molecules like terpenoids. The triterpene squalene has a variety of biotechnological uses and is the precursor to a diverse array of triterpenoids, but we currently lack a sustainable strategy to produce large quantities for industrial applications. Here, we further establish engineered plants as a platform for production of squalene through pathway re-targeting and membrane scaffolding. The squalene biosynthetic pathway, which natively resides in the cytosol and endoplasmic reticulum, was re-targeted to plastids, where screening of diverse variants of enzymes at key steps improved squalene yields. The highest yielding enzymes were used to create biosynthetic scaffolds on co-engineered, cytosolic lipid droplets, resulting in squalene yields up to 0.58 mg/gFW or 318% higher than a cytosolic pathway without scaffolding during transient expression. These scaffolds were also re-targeted to plastids where they associated with membranes throughout, including the formation of plastoglobules or plastidial lipid droplets. Plastid scaffolding ameliorated the negative effects of squalene biosynthesis and showed up to 345% higher rates of photosynthesis than without scaffolding. This study establishes a platform for engineering the production of squalene in plants, providing the opportunity to expand future work into production of higher-value triterpenoids.
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spelling pubmed-92080172022-06-21 Pathway Engineering, Re-targeting, and Synthetic Scaffolding Improve the Production of Squalene in Plants Bibik, Jacob D. Weraduwage, Sarathi M. Banerjee, Aparajita Robertson, Ka’shawn Espinoza-Corral, Roberto Sharkey, Thomas D. Lundquist, Peter K. Hamberger, Björn R. ACS Synth Biol [Image: see text] Plants are increasingly becoming an option for sustainable bioproduction of chemicals and complex molecules like terpenoids. The triterpene squalene has a variety of biotechnological uses and is the precursor to a diverse array of triterpenoids, but we currently lack a sustainable strategy to produce large quantities for industrial applications. Here, we further establish engineered plants as a platform for production of squalene through pathway re-targeting and membrane scaffolding. The squalene biosynthetic pathway, which natively resides in the cytosol and endoplasmic reticulum, was re-targeted to plastids, where screening of diverse variants of enzymes at key steps improved squalene yields. The highest yielding enzymes were used to create biosynthetic scaffolds on co-engineered, cytosolic lipid droplets, resulting in squalene yields up to 0.58 mg/gFW or 318% higher than a cytosolic pathway without scaffolding during transient expression. These scaffolds were also re-targeted to plastids where they associated with membranes throughout, including the formation of plastoglobules or plastidial lipid droplets. Plastid scaffolding ameliorated the negative effects of squalene biosynthesis and showed up to 345% higher rates of photosynthesis than without scaffolding. This study establishes a platform for engineering the production of squalene in plants, providing the opportunity to expand future work into production of higher-value triterpenoids. American Chemical Society 2022-05-13 2022-06-17 /pmc/articles/PMC9208017/ /pubmed/35549088 http://dx.doi.org/10.1021/acssynbio.2c00051 Text en © 2022 The Authors. Published by American Chemical Society https://creativecommons.org/licenses/by/4.0/Permits the broadest form of re-use including for commercial purposes, provided that author attribution and integrity are maintained (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Bibik, Jacob D.
Weraduwage, Sarathi M.
Banerjee, Aparajita
Robertson, Ka’shawn
Espinoza-Corral, Roberto
Sharkey, Thomas D.
Lundquist, Peter K.
Hamberger, Björn R.
Pathway Engineering, Re-targeting, and Synthetic Scaffolding Improve the Production of Squalene in Plants
title Pathway Engineering, Re-targeting, and Synthetic Scaffolding Improve the Production of Squalene in Plants
title_full Pathway Engineering, Re-targeting, and Synthetic Scaffolding Improve the Production of Squalene in Plants
title_fullStr Pathway Engineering, Re-targeting, and Synthetic Scaffolding Improve the Production of Squalene in Plants
title_full_unstemmed Pathway Engineering, Re-targeting, and Synthetic Scaffolding Improve the Production of Squalene in Plants
title_short Pathway Engineering, Re-targeting, and Synthetic Scaffolding Improve the Production of Squalene in Plants
title_sort pathway engineering, re-targeting, and synthetic scaffolding improve the production of squalene in plants
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9208017/
https://www.ncbi.nlm.nih.gov/pubmed/35549088
http://dx.doi.org/10.1021/acssynbio.2c00051
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