Improving Microbial Biogasoline Production in <named-content content-type="genus-species">Escherichia coli</named-content> Using Tolerance Engineering

ABSTRACT Engineering microbial hosts for the production of fungible fuels requires mitigation of limitations posed on the production capacity. One such limitation arises from the inherent toxicity of solvent-like biofuel compounds to production strains, such as Escherichia coli. Here we show the imp...

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Autores principales:	Jee Loon Foo, Heather M. Jensen, Robert H. Dahl, Kevin George, Jay D. Keasling, Taek Soon Lee, Susanna Leong, Aindrila Mukhopadhyay
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2014
Materias:	Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/f4f6aa8b2f174502ac528b49695d8004
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spelling	oai:doaj.org-article:f4f6aa8b2f174502ac528b49695d80042021-11-15T15:47:03ZImproving Microbial Biogasoline Production in <named-content content-type="genus-species">Escherichia coli</named-content> Using Tolerance Engineering10.1128/mBio.01932-142150-7511https://doaj.org/article/f4f6aa8b2f174502ac528b49695d80042014-12-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.01932-14https://doaj.org/toc/2150-7511ABSTRACT Engineering microbial hosts for the production of fungible fuels requires mitigation of limitations posed on the production capacity. One such limitation arises from the inherent toxicity of solvent-like biofuel compounds to production strains, such as Escherichia coli. Here we show the importance of host engineering for the production of short-chain alcohols by studying the overexpression of genes upregulated in response to exogenous isopentenol. Using systems biology data, we selected 40 genes that were upregulated following isopentenol exposure and subsequently overexpressed them in E. coli. Overexpression of several of these candidates improved tolerance to exogenously added isopentenol. Genes conferring isopentenol tolerance phenotypes belonged to diverse functional groups, such as oxidative stress response (soxS, fpr, and nrdH), general stress response (metR, yqhD, and gidB), heat shock-related response (ibpA), and transport (mdlB). To determine if these genes could also improve isopentenol production, we coexpressed the tolerance-enhancing genes individually with an isopentenol production pathway. Our data show that expression of 6 of the 8 candidates improved the production of isopentenol in E. coli, with the methionine biosynthesis regulator MetR improving the titer for isopentenol production by 55%. Additionally, expression of MdlB, an ABC transporter, facilitated a 12% improvement in isopentenol production. To our knowledge, MdlB is the first example of a transporter that can be used to improve production of a short-chain alcohol and provides a valuable new avenue for host engineering in biogasoline production. IMPORTANCE The use of microbial host platforms for the production of bulk commodities, such as chemicals and fuels, is now a focus of many biotechnology efforts. Many of these compounds are inherently toxic to the host microbe, which in turn places a limit on production despite efforts to optimize the bioconversion pathways. In order to achieve economically viable production levels, it is also necessary to engineer production strains with improved tolerance to these compounds. We demonstrate that microbial tolerance engineering using transcriptomics data can also identify targets that improve production. Our results include an exporter and a methionine biosynthesis regulator that improve isopentenol production, providing a starting point to further engineer the host for biogasoline production.Jee Loon FooHeather M. JensenRobert H. DahlKevin GeorgeJay D. KeaslingTaek Soon LeeSusanna LeongAindrila MukhopadhyayAmerican Society for MicrobiologyarticleMicrobiologyQR1-502ENmBio, Vol 5, Iss 6 (2014)
institution	DOAJ
collection	DOAJ
language	EN
topic	Microbiology QR1-502
spellingShingle	Microbiology QR1-502 Jee Loon Foo Heather M. Jensen Robert H. Dahl Kevin George Jay D. Keasling Taek Soon Lee Susanna Leong Aindrila Mukhopadhyay Improving Microbial Biogasoline Production in <named-content content-type="genus-species">Escherichia coli</named-content> Using Tolerance Engineering
description	ABSTRACT Engineering microbial hosts for the production of fungible fuels requires mitigation of limitations posed on the production capacity. One such limitation arises from the inherent toxicity of solvent-like biofuel compounds to production strains, such as Escherichia coli. Here we show the importance of host engineering for the production of short-chain alcohols by studying the overexpression of genes upregulated in response to exogenous isopentenol. Using systems biology data, we selected 40 genes that were upregulated following isopentenol exposure and subsequently overexpressed them in E. coli. Overexpression of several of these candidates improved tolerance to exogenously added isopentenol. Genes conferring isopentenol tolerance phenotypes belonged to diverse functional groups, such as oxidative stress response (soxS, fpr, and nrdH), general stress response (metR, yqhD, and gidB), heat shock-related response (ibpA), and transport (mdlB). To determine if these genes could also improve isopentenol production, we coexpressed the tolerance-enhancing genes individually with an isopentenol production pathway. Our data show that expression of 6 of the 8 candidates improved the production of isopentenol in E. coli, with the methionine biosynthesis regulator MetR improving the titer for isopentenol production by 55%. Additionally, expression of MdlB, an ABC transporter, facilitated a 12% improvement in isopentenol production. To our knowledge, MdlB is the first example of a transporter that can be used to improve production of a short-chain alcohol and provides a valuable new avenue for host engineering in biogasoline production. IMPORTANCE The use of microbial host platforms for the production of bulk commodities, such as chemicals and fuels, is now a focus of many biotechnology efforts. Many of these compounds are inherently toxic to the host microbe, which in turn places a limit on production despite efforts to optimize the bioconversion pathways. In order to achieve economically viable production levels, it is also necessary to engineer production strains with improved tolerance to these compounds. We demonstrate that microbial tolerance engineering using transcriptomics data can also identify targets that improve production. Our results include an exporter and a methionine biosynthesis regulator that improve isopentenol production, providing a starting point to further engineer the host for biogasoline production.
format	article
author	Jee Loon Foo Heather M. Jensen Robert H. Dahl Kevin George Jay D. Keasling Taek Soon Lee Susanna Leong Aindrila Mukhopadhyay
author_facet	Jee Loon Foo Heather M. Jensen Robert H. Dahl Kevin George Jay D. Keasling Taek Soon Lee Susanna Leong Aindrila Mukhopadhyay
author_sort	Jee Loon Foo
title	Improving Microbial Biogasoline Production in <named-content content-type="genus-species">Escherichia coli</named-content> Using Tolerance Engineering
title_short	Improving Microbial Biogasoline Production in <named-content content-type="genus-species">Escherichia coli</named-content> Using Tolerance Engineering
title_full	Improving Microbial Biogasoline Production in <named-content content-type="genus-species">Escherichia coli</named-content> Using Tolerance Engineering
title_fullStr	Improving Microbial Biogasoline Production in <named-content content-type="genus-species">Escherichia coli</named-content> Using Tolerance Engineering
title_full_unstemmed	Improving Microbial Biogasoline Production in <named-content content-type="genus-species">Escherichia coli</named-content> Using Tolerance Engineering
title_sort	improving microbial biogasoline production in <named-content content-type="genus-species">escherichia coli</named-content> using tolerance engineering
publisher	American Society for Microbiology
publishDate	2014
url	https://doaj.org/article/f4f6aa8b2f174502ac528b49695d8004
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Improving Microbial Biogasoline Production in <named-content content-type="genus-species">Escherichia coli</named-content> Using Tolerance Engineering

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