Complete Biosynthesis of Anthocyanins Using <italic toggle="yes">E. coli</italic> Polycultures

ABSTRACT Fermentation-based chemical production strategies provide a feasible route for the rapid, safe, and sustainable production of a wide variety of important chemical products, ranging from fuels to pharmaceuticals. These strategies have yet to find wide industrial utilization due to their inab...

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Autores principales:	J. Andrew Jones, Victoria R. Vernacchio, Shannon M. Collins, Abhijit N. Shirke, Yu Xiu, Jacob A. Englaender, Brady F. Cress, Catherine C. McCutcheon, Robert J. Linhardt, Richard A. Gross, Mattheos A. G. Koffas
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2017
Materias:	Escherichia coli anthocyanins coculture de novo flavonoids pelargonidin 3-O-glucoside Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/e8e823118eb74a3091c85258ec13f55c
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spelling	oai:doaj.org-article:e8e823118eb74a3091c85258ec13f55c2021-11-15T15:51:29ZComplete Biosynthesis of Anthocyanins Using <italic toggle="yes">E. coli</italic> Polycultures10.1128/mBio.00621-172150-7511https://doaj.org/article/e8e823118eb74a3091c85258ec13f55c2017-07-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00621-17https://doaj.org/toc/2150-7511ABSTRACT Fermentation-based chemical production strategies provide a feasible route for the rapid, safe, and sustainable production of a wide variety of important chemical products, ranging from fuels to pharmaceuticals. These strategies have yet to find wide industrial utilization due to their inability to economically compete with traditional extraction and chemical production methods. Here, we engineer for the first time the complex microbial biosynthesis of an anthocyanin plant natural product, starting from sugar. This was accomplished through the development of a synthetic, 4-strain Escherichia coli polyculture collectively expressing 15 exogenous or modified pathway enzymes from diverse plants and other microbes. This synthetic consortium-based approach enables the functional expression and connection of lengthy pathways while effectively managing the accompanying metabolic burden. The de novo production of specific anthocyanin molecules, such as calistephin, has been an elusive metabolic engineering target for over a decade. The utilization of our polyculture strategy affords milligram-per-liter production titers. This study also lays the groundwork for significant advances in strain and process design toward the development of cost-competitive biochemical production hosts through nontraditional methodologies. IMPORTANCE To efficiently express active extensive recombinant pathways with high flux in microbial hosts requires careful balance and allocation of metabolic resources such as ATP, reducing equivalents, and malonyl coenzyme A (malonyl-CoA), as well as various other pathway-dependent cofactors and precursors. To address this issue, we report the design, characterization, and implementation of the first synthetic 4-strain polyculture. Division of the overexpression of 15 enzymes and transcription factors over 4 independent strain modules allowed for the division of metabolic burden and for independent strain optimization for module-specific metabolite needs. This study represents the most complex synthetic consortia constructed to date for metabolic engineering applications and provides a new paradigm in metabolic engineering for the reconstitution of extensive metabolic pathways in nonnative hosts.J. Andrew JonesVictoria R. VernacchioShannon M. CollinsAbhijit N. ShirkeYu XiuJacob A. EnglaenderBrady F. CressCatherine C. McCutcheonRobert J. LinhardtRichard A. GrossMattheos A. G. KoffasAmerican Society for MicrobiologyarticleEscherichia colianthocyaninscoculturede novoflavonoidspelargonidin 3-O-glucosideMicrobiologyQR1-502ENmBio, Vol 8, Iss 3 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Escherichia coli anthocyanins coculture de novo flavonoids pelargonidin 3-O-glucoside Microbiology QR1-502
spellingShingle	Escherichia coli anthocyanins coculture de novo flavonoids pelargonidin 3-O-glucoside Microbiology QR1-502 J. Andrew Jones Victoria R. Vernacchio Shannon M. Collins Abhijit N. Shirke Yu Xiu Jacob A. Englaender Brady F. Cress Catherine C. McCutcheon Robert J. Linhardt Richard A. Gross Mattheos A. G. Koffas Complete Biosynthesis of Anthocyanins Using <italic toggle="yes">E. coli</italic> Polycultures
description	ABSTRACT Fermentation-based chemical production strategies provide a feasible route for the rapid, safe, and sustainable production of a wide variety of important chemical products, ranging from fuels to pharmaceuticals. These strategies have yet to find wide industrial utilization due to their inability to economically compete with traditional extraction and chemical production methods. Here, we engineer for the first time the complex microbial biosynthesis of an anthocyanin plant natural product, starting from sugar. This was accomplished through the development of a synthetic, 4-strain Escherichia coli polyculture collectively expressing 15 exogenous or modified pathway enzymes from diverse plants and other microbes. This synthetic consortium-based approach enables the functional expression and connection of lengthy pathways while effectively managing the accompanying metabolic burden. The de novo production of specific anthocyanin molecules, such as calistephin, has been an elusive metabolic engineering target for over a decade. The utilization of our polyculture strategy affords milligram-per-liter production titers. This study also lays the groundwork for significant advances in strain and process design toward the development of cost-competitive biochemical production hosts through nontraditional methodologies. IMPORTANCE To efficiently express active extensive recombinant pathways with high flux in microbial hosts requires careful balance and allocation of metabolic resources such as ATP, reducing equivalents, and malonyl coenzyme A (malonyl-CoA), as well as various other pathway-dependent cofactors and precursors. To address this issue, we report the design, characterization, and implementation of the first synthetic 4-strain polyculture. Division of the overexpression of 15 enzymes and transcription factors over 4 independent strain modules allowed for the division of metabolic burden and for independent strain optimization for module-specific metabolite needs. This study represents the most complex synthetic consortia constructed to date for metabolic engineering applications and provides a new paradigm in metabolic engineering for the reconstitution of extensive metabolic pathways in nonnative hosts.
format	article
author	J. Andrew Jones Victoria R. Vernacchio Shannon M. Collins Abhijit N. Shirke Yu Xiu Jacob A. Englaender Brady F. Cress Catherine C. McCutcheon Robert J. Linhardt Richard A. Gross Mattheos A. G. Koffas
author_facet	J. Andrew Jones Victoria R. Vernacchio Shannon M. Collins Abhijit N. Shirke Yu Xiu Jacob A. Englaender Brady F. Cress Catherine C. McCutcheon Robert J. Linhardt Richard A. Gross Mattheos A. G. Koffas
author_sort	J. Andrew Jones
title	Complete Biosynthesis of Anthocyanins Using <italic toggle="yes">E. coli</italic> Polycultures
title_short	Complete Biosynthesis of Anthocyanins Using <italic toggle="yes">E. coli</italic> Polycultures
title_full	Complete Biosynthesis of Anthocyanins Using <italic toggle="yes">E. coli</italic> Polycultures
title_fullStr	Complete Biosynthesis of Anthocyanins Using <italic toggle="yes">E. coli</italic> Polycultures
title_full_unstemmed	Complete Biosynthesis of Anthocyanins Using <italic toggle="yes">E. coli</italic> Polycultures
title_sort	complete biosynthesis of anthocyanins using <italic toggle="yes">e. coli</italic> polycultures
publisher	American Society for Microbiology
publishDate	2017
url	https://doaj.org/article/e8e823118eb74a3091c85258ec13f55c
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Complete Biosynthesis of Anthocyanins Using <italic toggle="yes">E. coli</italic> Polycultures

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