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Combining Genotype Improvement and Statistical Media Optimization for Isoprenoid Production in E. coli
Isoprenoids are a large and diverse class of compounds that includes many high value natural products and are thus in great demand. To meet the increasing demand for isoprenoid compounds, metabolic engineering of microbes has been used to produce isoprenoids in an economical and sustainable manner....
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Public Library of Science
2013
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3790805/ https://www.ncbi.nlm.nih.gov/pubmed/24124471 http://dx.doi.org/10.1371/journal.pone.0075164 |
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author | Zhang, Congqiang Chen, Xixian Zou, Ruiyang Zhou, Kang Stephanopoulos, Gregory Too, Heng-Phon |
author_facet | Zhang, Congqiang Chen, Xixian Zou, Ruiyang Zhou, Kang Stephanopoulos, Gregory Too, Heng-Phon |
author_sort | Zhang, Congqiang |
collection | PubMed |
description | Isoprenoids are a large and diverse class of compounds that includes many high value natural products and are thus in great demand. To meet the increasing demand for isoprenoid compounds, metabolic engineering of microbes has been used to produce isoprenoids in an economical and sustainable manner. To achieve high isoprenoid yields using this technology, the availability of metabolic precursors feeding the deoxyxylulose phosphate (DXP) pathway, responsible for isoprenoid biosynthesis, has to be optimized. In this study, phosphoenolpyruvate, a vital DXP pathway precursor, was enriched by deleting the genes encoding the carbohydrate phosphotransferase system (PTS) in E. coli. Production of lycopene (a C40 isoprenoid) was maximized by optimizing growth medium and culture conditions. In optimized conditions, the lycopene yield from PTS mutant was seven fold higher than that obtained from the wild type strain. This resulted in the highest reported specific yield of lycopene produced from the DXP pathway in E. coli to date (20,000 µg/g dry cell weight). Both the copy number of the plasmid encoding the lycopene biosynthetic genes and the expression were found to be increased in the optimized media. Deletion of PTS together with a similar optimization strategy was also successful in enhancing the production of amorpha-1,4-diene, a distinct C15 isoprenoid, suggesting that the approaches developed herein can be generally applied to optimize production of other isoprenoids. |
format | Online Article Text |
id | pubmed-3790805 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-37908052013-10-11 Combining Genotype Improvement and Statistical Media Optimization for Isoprenoid Production in E. coli Zhang, Congqiang Chen, Xixian Zou, Ruiyang Zhou, Kang Stephanopoulos, Gregory Too, Heng-Phon PLoS One Research Article Isoprenoids are a large and diverse class of compounds that includes many high value natural products and are thus in great demand. To meet the increasing demand for isoprenoid compounds, metabolic engineering of microbes has been used to produce isoprenoids in an economical and sustainable manner. To achieve high isoprenoid yields using this technology, the availability of metabolic precursors feeding the deoxyxylulose phosphate (DXP) pathway, responsible for isoprenoid biosynthesis, has to be optimized. In this study, phosphoenolpyruvate, a vital DXP pathway precursor, was enriched by deleting the genes encoding the carbohydrate phosphotransferase system (PTS) in E. coli. Production of lycopene (a C40 isoprenoid) was maximized by optimizing growth medium and culture conditions. In optimized conditions, the lycopene yield from PTS mutant was seven fold higher than that obtained from the wild type strain. This resulted in the highest reported specific yield of lycopene produced from the DXP pathway in E. coli to date (20,000 µg/g dry cell weight). Both the copy number of the plasmid encoding the lycopene biosynthetic genes and the expression were found to be increased in the optimized media. Deletion of PTS together with a similar optimization strategy was also successful in enhancing the production of amorpha-1,4-diene, a distinct C15 isoprenoid, suggesting that the approaches developed herein can be generally applied to optimize production of other isoprenoids. Public Library of Science 2013-10-04 /pmc/articles/PMC3790805/ /pubmed/24124471 http://dx.doi.org/10.1371/journal.pone.0075164 Text en © 2013 Zhang et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Zhang, Congqiang Chen, Xixian Zou, Ruiyang Zhou, Kang Stephanopoulos, Gregory Too, Heng-Phon Combining Genotype Improvement and Statistical Media Optimization for Isoprenoid Production in E. coli |
title | Combining Genotype Improvement and Statistical Media Optimization for Isoprenoid Production in E. coli
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title_full | Combining Genotype Improvement and Statistical Media Optimization for Isoprenoid Production in E. coli
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title_fullStr | Combining Genotype Improvement and Statistical Media Optimization for Isoprenoid Production in E. coli
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title_full_unstemmed | Combining Genotype Improvement and Statistical Media Optimization for Isoprenoid Production in E. coli
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title_short | Combining Genotype Improvement and Statistical Media Optimization for Isoprenoid Production in E. coli
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title_sort | combining genotype improvement and statistical media optimization for isoprenoid production in e. coli |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3790805/ https://www.ncbi.nlm.nih.gov/pubmed/24124471 http://dx.doi.org/10.1371/journal.pone.0075164 |
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