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....
| Main Authors: | , , , , , |
|---|---|
| Other Authors: | |
| Format: | Article |
| Language: | en_US |
| Published: |
Public Library of Science
2014
|
| Online Access: | http://hdl.handle.net/1721.1/83863 https://orcid.org/0000-0001-6909-4568 |
| _version_ | 1826217741476954112 |
|---|---|
| author | Zhang, Congqiang Chen, Xixian Zou, Ruiyang Zhou, Kang Stephanopoulos, Gregory Too, Heng-Phon |
| author2 | Massachusetts Institute of Technology. Department of Chemical Engineering |
| author_facet | Massachusetts Institute of Technology. Department of Chemical Engineering Zhang, Congqiang Chen, Xixian Zou, Ruiyang Zhou, Kang Stephanopoulos, Gregory Too, Heng-Phon |
| author_sort | Zhang, Congqiang |
| collection | MIT |
| 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. |
| first_indexed | 2024-09-23T17:08:25Z |
| format | Article |
| id | mit-1721.1/83863 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T17:08:25Z |
| publishDate | 2014 |
| publisher | Public Library of Science |
| record_format | dspace |
| spelling | mit-1721.1/838632022-09-29T23:55:49Z 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 Massachusetts Institute of Technology. Department of Chemical Engineering Singapore-MIT Alliance in Research and Technology (SMART) Zhang, Congqiang Chen, Xixian Zhou, Kang Stephanopoulos, Gregory 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. Singapore-MIT Alliance 2014-01-10T18:46:03Z 2014-01-10T18:46:03Z 2013-10 2013-01 Article http://purl.org/eprint/type/JournalArticle 1932-6203 http://hdl.handle.net/1721.1/83863 Zhang, Congqiang, Xixian Chen, Ruiyang Zou, Kang Zhou, Gregory Stephanopoulos, and Heng-Phon Too. “Combining Genotype Improvement and Statistical Media Optimization for Isoprenoid Production in E. coli.” Edited by Arnold Driessen. PLoS ONE 8, no. 10 (October 4, 2013): e75164. https://orcid.org/0000-0001-6909-4568 en_US http://dx.doi.org/10.1371/journal.pone.0075164 PLoS ONE http://creativecommons.org/licenses/by/2.5/ application/pdf Public Library of Science PLoS |
| spellingShingle | 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 |
| title_full | Combining Genotype Improvement and Statistical Media Optimization for Isoprenoid Production in E. coli |
| title_fullStr | Combining Genotype Improvement and Statistical Media Optimization for Isoprenoid Production in E. coli |
| title_full_unstemmed | Combining Genotype Improvement and Statistical Media Optimization for Isoprenoid Production in E. coli |
| title_short | Combining Genotype Improvement and Statistical Media Optimization for Isoprenoid Production in E. coli |
| title_sort | combining genotype improvement and statistical media optimization for isoprenoid production in e coli |
| url | http://hdl.handle.net/1721.1/83863 https://orcid.org/0000-0001-6909-4568 |
| work_keys_str_mv | AT zhangcongqiang combininggenotypeimprovementandstatisticalmediaoptimizationforisoprenoidproductioninecoli AT chenxixian combininggenotypeimprovementandstatisticalmediaoptimizationforisoprenoidproductioninecoli AT zouruiyang combininggenotypeimprovementandstatisticalmediaoptimizationforisoprenoidproductioninecoli AT zhoukang combininggenotypeimprovementandstatisticalmediaoptimizationforisoprenoidproductioninecoli AT stephanopoulosgregory combininggenotypeimprovementandstatisticalmediaoptimizationforisoprenoidproductioninecoli AT toohengphon combininggenotypeimprovementandstatisticalmediaoptimizationforisoprenoidproductioninecoli |