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Improving Microbial Biogasoline Production in Escherichia coli Using Tolerance Engineering
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 o...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Society of Microbiology
2014
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4222104/ https://www.ncbi.nlm.nih.gov/pubmed/25370492 http://dx.doi.org/10.1128/mBio.01932-14 |
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author | Foo, Jee Loon Jensen, Heather M. Dahl, Robert H. George, Kevin Keasling, Jay D. Lee, Taek Soon Leong, Susanna Mukhopadhyay, Aindrila |
author_facet | Foo, Jee Loon Jensen, Heather M. Dahl, Robert H. George, Kevin Keasling, Jay D. Lee, Taek Soon Leong, Susanna Mukhopadhyay, Aindrila |
author_sort | Foo, Jee Loon |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-4222104 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | American Society of Microbiology |
record_format | MEDLINE/PubMed |
spelling | pubmed-42221042014-11-06 Improving Microbial Biogasoline Production in Escherichia coli Using Tolerance Engineering Foo, Jee Loon Jensen, Heather M. Dahl, Robert H. George, Kevin Keasling, Jay D. Lee, Taek Soon Leong, Susanna Mukhopadhyay, Aindrila mBio Research Article 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. American Society of Microbiology 2014-11-04 /pmc/articles/PMC4222104/ /pubmed/25370492 http://dx.doi.org/10.1128/mBio.01932-14 Text en Copyright © 2014 Foo et al. http://creativecommons.org/licenses/by-nc-sa/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution-Noncommercial-ShareAlike 3.0 Unported license (http://creativecommons.org/licenses/by-nc-sa/3.0/) , which permits unrestricted noncommercial use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Article Foo, Jee Loon Jensen, Heather M. Dahl, Robert H. George, Kevin Keasling, Jay D. Lee, Taek Soon Leong, Susanna Mukhopadhyay, Aindrila Improving Microbial Biogasoline Production in Escherichia coli Using Tolerance Engineering |
title | Improving Microbial Biogasoline Production in Escherichia coli Using Tolerance Engineering |
title_full | Improving Microbial Biogasoline Production in Escherichia coli Using Tolerance Engineering |
title_fullStr | Improving Microbial Biogasoline Production in Escherichia coli Using Tolerance Engineering |
title_full_unstemmed | Improving Microbial Biogasoline Production in Escherichia coli Using Tolerance Engineering |
title_short | Improving Microbial Biogasoline Production in Escherichia coli Using Tolerance Engineering |
title_sort | improving microbial biogasoline production in escherichia coli using tolerance engineering |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4222104/ https://www.ncbi.nlm.nih.gov/pubmed/25370492 http://dx.doi.org/10.1128/mBio.01932-14 |
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