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Plant-Derived Cell-Free Biofactories for the Production of Secondary Metabolites

Cell-free expression systems enable the production of proteins and metabolites within a few hours or days. Removing the cellular context while maintaining the protein biosynthesis apparatus provides an open system that allows metabolic pathways to be installed and optimized by expressing different n...

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Autores principales: Buntru, Matthias, Hahnengress, Nils, Croon, Alexander, Schillberg, Stefan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8832058/
https://www.ncbi.nlm.nih.gov/pubmed/35154185
http://dx.doi.org/10.3389/fpls.2021.794999
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author Buntru, Matthias
Hahnengress, Nils
Croon, Alexander
Schillberg, Stefan
author_facet Buntru, Matthias
Hahnengress, Nils
Croon, Alexander
Schillberg, Stefan
author_sort Buntru, Matthias
collection PubMed
description Cell-free expression systems enable the production of proteins and metabolites within a few hours or days. Removing the cellular context while maintaining the protein biosynthesis apparatus provides an open system that allows metabolic pathways to be installed and optimized by expressing different numbers and combinations of enzymes. This facilitates the synthesis of secondary metabolites that are difficult to produce in cell-based systems because they are toxic to the host cell or immediately converted into downstream products. Recently, we developed a cell-free lysate derived from tobacco BY-2 cell suspension cultures for the production of recombinant proteins. This system is remarkably productive, achieving yields of up to 3 mg/mL in a one-pot in vitro transcription–translation reaction and contains highly active energy and cofactor regeneration pathways. Here, we demonstrate for the first time that the BY-2 cell-free lysate also allows the efficient production of several classes of secondary metabolites. As case studies, we synthesized lycopene, indigoidine, betanin, and betaxanthins, which are useful in the food, cosmetic, textile, and pharmaceutical industries. Production was achieved by the co-expression of up to three metabolic enzymes. For all four products, we achieved medium to high yields. However, the yield of betanin (555 μg/mL) was outstanding, exceeding the level reported in yeast cells by a factor of more than 30. Our results show that the BY-2 cell-free lysate is suitable not only for the verification and optimization of metabolic pathways, but also for the efficient production of small to medium quantities of secondary metabolites.
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spelling pubmed-88320582022-02-12 Plant-Derived Cell-Free Biofactories for the Production of Secondary Metabolites Buntru, Matthias Hahnengress, Nils Croon, Alexander Schillberg, Stefan Front Plant Sci Plant Science Cell-free expression systems enable the production of proteins and metabolites within a few hours or days. Removing the cellular context while maintaining the protein biosynthesis apparatus provides an open system that allows metabolic pathways to be installed and optimized by expressing different numbers and combinations of enzymes. This facilitates the synthesis of secondary metabolites that are difficult to produce in cell-based systems because they are toxic to the host cell or immediately converted into downstream products. Recently, we developed a cell-free lysate derived from tobacco BY-2 cell suspension cultures for the production of recombinant proteins. This system is remarkably productive, achieving yields of up to 3 mg/mL in a one-pot in vitro transcription–translation reaction and contains highly active energy and cofactor regeneration pathways. Here, we demonstrate for the first time that the BY-2 cell-free lysate also allows the efficient production of several classes of secondary metabolites. As case studies, we synthesized lycopene, indigoidine, betanin, and betaxanthins, which are useful in the food, cosmetic, textile, and pharmaceutical industries. Production was achieved by the co-expression of up to three metabolic enzymes. For all four products, we achieved medium to high yields. However, the yield of betanin (555 μg/mL) was outstanding, exceeding the level reported in yeast cells by a factor of more than 30. Our results show that the BY-2 cell-free lysate is suitable not only for the verification and optimization of metabolic pathways, but also for the efficient production of small to medium quantities of secondary metabolites. Frontiers Media S.A. 2022-01-28 /pmc/articles/PMC8832058/ /pubmed/35154185 http://dx.doi.org/10.3389/fpls.2021.794999 Text en Copyright © 2022 Buntru, Hahnengress, Croon and Schillberg. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Plant Science
Buntru, Matthias
Hahnengress, Nils
Croon, Alexander
Schillberg, Stefan
Plant-Derived Cell-Free Biofactories for the Production of Secondary Metabolites
title Plant-Derived Cell-Free Biofactories for the Production of Secondary Metabolites
title_full Plant-Derived Cell-Free Biofactories for the Production of Secondary Metabolites
title_fullStr Plant-Derived Cell-Free Biofactories for the Production of Secondary Metabolites
title_full_unstemmed Plant-Derived Cell-Free Biofactories for the Production of Secondary Metabolites
title_short Plant-Derived Cell-Free Biofactories for the Production of Secondary Metabolites
title_sort plant-derived cell-free biofactories for the production of secondary metabolites
topic Plant Science
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8832058/
https://www.ncbi.nlm.nih.gov/pubmed/35154185
http://dx.doi.org/10.3389/fpls.2021.794999
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