Development of a Synthetic 3‑Dehydroshikimate Biosensor in Escherichia coli for Metabolite Monitoring and Genetic Screening
Biosensors for target metabolites provide powerful high-throughput screening tools to obtain high-performing strains. However, well-characterized metabolite-sensing modules are often unavailable and limit rapid access to the robust biosensors with successful applications. In this study, we developed...
Gespeichert in:
Veröffentlicht in: | ACS synthetic biology 2019-02, Vol.8 (2), p.297-306 |
---|---|
Hauptverfasser: | , , , , , , , |
Format: | Artikel |
Sprache: | eng |
Schlagworte: | |
Online-Zugang: | Volltext |
Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
container_end_page | 306 |
---|---|
container_issue | 2 |
container_start_page | 297 |
container_title | ACS synthetic biology |
container_volume | 8 |
creator | Li, Liangpo Tu, Ran Song, Guotian Cheng, Jie Chen, Wujiu Li, Lin Wang, Lixian Wang, Qinhong |
description | Biosensors for target metabolites provide powerful high-throughput screening tools to obtain high-performing strains. However, well-characterized metabolite-sensing modules are often unavailable and limit rapid access to the robust biosensors with successful applications. In this study, we developed a strategy of transcriptome-assisted metabolite-sensing (TAMES) to identify the target metabolite-sensing module based on selectively comparative transcriptome analysis between the target metabolite producing and nonproducing strains and a subsequent quantative reverse transcription (RT-qPCR) evaluation. The strategy was applied to identify the sensing module cusR that responds positively to the metabolite 3-dehydroshikimate (DHS) and proved it was effective to narrow down the candidates. We further constructed the cusR-based synthetic biosensor and established the DHS biosensor-based high-throughput screening (HTS) platform to screen higher DHS-producing strains and successfully increased DHS production by more than 90%. This study demonstrated that the TAMES strategy was effective at exploiting the metabolite-sensing transcriptional regulator, and this could likely be extended to develop the biosensor-based HTS platforms for other molecules. |
doi_str_mv | 10.1021/acssynbio.8b00317 |
format | Article |
fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_2164102721</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>2164102721</sourcerecordid><originalsourceid>FETCH-LOGICAL-a339t-cf2e419d91ffcf1a39dcf5957ff8c4d2ed72a4bea01a259542677e3d87608f313</originalsourceid><addsrcrecordid>eNp9kM1OAjEcxBujEYI8gBfTo5fFdrufRwVEE4gH9Lzpdv91i0uL7WLCwcRX8BV9Eosg8WQvbSczk8wPoXNKBpSE9IoL5za6VGaQlYQwmh6hbkgTGsQkYcd_3h3Ud25B_IljFrPsFHUYSUieZVkXvY_gDRqzWoJusZGY4_lGtzW0SmD29fE5gnpTWeNq9aKWvAV8o4wD7YzFSuOxEzVYJWrFsTCNwtLrM2h56T_ePDNatcYq_Yy5rvAE9E_xXFgA7dUzdCJ546C_v3vo6Xb8OLwLpg-T--H1NOCM5W0gZAgRzaucSikk5SyvhIzzOJUyE1EVQpWGPCqBE8pDr0dhkqbAqixNSCYZZT10uetdWfO6BtcWS-UENA3XYNau8KwiDzUNt1a6swq_2lmQxcr65XZTUFJswRcH8MUevM9c7OvX5RKqQ-IXszcEO4PPFguzttqv_afwG92_k2g</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>2164102721</pqid></control><display><type>article</type><title>Development of a Synthetic 3‑Dehydroshikimate Biosensor in Escherichia coli for Metabolite Monitoring and Genetic Screening</title><source>MEDLINE</source><source>American Chemical Society Journals</source><creator>Li, Liangpo ; Tu, Ran ; Song, Guotian ; Cheng, Jie ; Chen, Wujiu ; Li, Lin ; Wang, Lixian ; Wang, Qinhong</creator><creatorcontrib>Li, Liangpo ; Tu, Ran ; Song, Guotian ; Cheng, Jie ; Chen, Wujiu ; Li, Lin ; Wang, Lixian ; Wang, Qinhong</creatorcontrib><description>Biosensors for target metabolites provide powerful high-throughput screening tools to obtain high-performing strains. However, well-characterized metabolite-sensing modules are often unavailable and limit rapid access to the robust biosensors with successful applications. In this study, we developed a strategy of transcriptome-assisted metabolite-sensing (TAMES) to identify the target metabolite-sensing module based on selectively comparative transcriptome analysis between the target metabolite producing and nonproducing strains and a subsequent quantative reverse transcription (RT-qPCR) evaluation. The strategy was applied to identify the sensing module cusR that responds positively to the metabolite 3-dehydroshikimate (DHS) and proved it was effective to narrow down the candidates. We further constructed the cusR-based synthetic biosensor and established the DHS biosensor-based high-throughput screening (HTS) platform to screen higher DHS-producing strains and successfully increased DHS production by more than 90%. This study demonstrated that the TAMES strategy was effective at exploiting the metabolite-sensing transcriptional regulator, and this could likely be extended to develop the biosensor-based HTS platforms for other molecules.</description><identifier>ISSN: 2161-5063</identifier><identifier>EISSN: 2161-5063</identifier><identifier>DOI: 10.1021/acssynbio.8b00317</identifier><identifier>PMID: 30609888</identifier><language>eng</language><publisher>United States: American Chemical Society</publisher><subject>Biosensing Techniques ; Escherichia coli - metabolism ; Gene Expression Regulation - genetics ; Metabolic Engineering - methods ; Shikimic Acid - analogs & derivatives ; Transcription Factors - metabolism ; Transcriptome - genetics</subject><ispartof>ACS synthetic biology, 2019-02, Vol.8 (2), p.297-306</ispartof><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-a339t-cf2e419d91ffcf1a39dcf5957ff8c4d2ed72a4bea01a259542677e3d87608f313</citedby><cites>FETCH-LOGICAL-a339t-cf2e419d91ffcf1a39dcf5957ff8c4d2ed72a4bea01a259542677e3d87608f313</cites><orcidid>0000-0003-0004-2721</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://pubs.acs.org/doi/pdf/10.1021/acssynbio.8b00317$$EPDF$$P50$$Gacs$$H</linktopdf><linktohtml>$$Uhttps://pubs.acs.org/doi/10.1021/acssynbio.8b00317$$EHTML$$P50$$Gacs$$H</linktohtml><link.rule.ids>314,780,784,2765,27076,27924,27925,56738,56788</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/30609888$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Li, Liangpo</creatorcontrib><creatorcontrib>Tu, Ran</creatorcontrib><creatorcontrib>Song, Guotian</creatorcontrib><creatorcontrib>Cheng, Jie</creatorcontrib><creatorcontrib>Chen, Wujiu</creatorcontrib><creatorcontrib>Li, Lin</creatorcontrib><creatorcontrib>Wang, Lixian</creatorcontrib><creatorcontrib>Wang, Qinhong</creatorcontrib><title>Development of a Synthetic 3‑Dehydroshikimate Biosensor in Escherichia coli for Metabolite Monitoring and Genetic Screening</title><title>ACS synthetic biology</title><addtitle>ACS Synth. Biol</addtitle><description>Biosensors for target metabolites provide powerful high-throughput screening tools to obtain high-performing strains. However, well-characterized metabolite-sensing modules are often unavailable and limit rapid access to the robust biosensors with successful applications. In this study, we developed a strategy of transcriptome-assisted metabolite-sensing (TAMES) to identify the target metabolite-sensing module based on selectively comparative transcriptome analysis between the target metabolite producing and nonproducing strains and a subsequent quantative reverse transcription (RT-qPCR) evaluation. The strategy was applied to identify the sensing module cusR that responds positively to the metabolite 3-dehydroshikimate (DHS) and proved it was effective to narrow down the candidates. We further constructed the cusR-based synthetic biosensor and established the DHS biosensor-based high-throughput screening (HTS) platform to screen higher DHS-producing strains and successfully increased DHS production by more than 90%. This study demonstrated that the TAMES strategy was effective at exploiting the metabolite-sensing transcriptional regulator, and this could likely be extended to develop the biosensor-based HTS platforms for other molecules.</description><subject>Biosensing Techniques</subject><subject>Escherichia coli - metabolism</subject><subject>Gene Expression Regulation - genetics</subject><subject>Metabolic Engineering - methods</subject><subject>Shikimic Acid - analogs & derivatives</subject><subject>Transcription Factors - metabolism</subject><subject>Transcriptome - genetics</subject><issn>2161-5063</issn><issn>2161-5063</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2019</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kM1OAjEcxBujEYI8gBfTo5fFdrufRwVEE4gH9Lzpdv91i0uL7WLCwcRX8BV9Eosg8WQvbSczk8wPoXNKBpSE9IoL5za6VGaQlYQwmh6hbkgTGsQkYcd_3h3Ud25B_IljFrPsFHUYSUieZVkXvY_gDRqzWoJusZGY4_lGtzW0SmD29fE5gnpTWeNq9aKWvAV8o4wD7YzFSuOxEzVYJWrFsTCNwtLrM2h56T_ePDNatcYq_Yy5rvAE9E_xXFgA7dUzdCJ546C_v3vo6Xb8OLwLpg-T--H1NOCM5W0gZAgRzaucSikk5SyvhIzzOJUyE1EVQpWGPCqBE8pDr0dhkqbAqixNSCYZZT10uetdWfO6BtcWS-UENA3XYNau8KwiDzUNt1a6swq_2lmQxcr65XZTUFJswRcH8MUevM9c7OvX5RKqQ-IXszcEO4PPFguzttqv_afwG92_k2g</recordid><startdate>20190215</startdate><enddate>20190215</enddate><creator>Li, Liangpo</creator><creator>Tu, Ran</creator><creator>Song, Guotian</creator><creator>Cheng, Jie</creator><creator>Chen, Wujiu</creator><creator>Li, Lin</creator><creator>Wang, Lixian</creator><creator>Wang, Qinhong</creator><general>American Chemical Society</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><orcidid>https://orcid.org/0000-0003-0004-2721</orcidid></search><sort><creationdate>20190215</creationdate><title>Development of a Synthetic 3‑Dehydroshikimate Biosensor in Escherichia coli for Metabolite Monitoring and Genetic Screening</title><author>Li, Liangpo ; Tu, Ran ; Song, Guotian ; Cheng, Jie ; Chen, Wujiu ; Li, Lin ; Wang, Lixian ; Wang, Qinhong</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-a339t-cf2e419d91ffcf1a39dcf5957ff8c4d2ed72a4bea01a259542677e3d87608f313</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2019</creationdate><topic>Biosensing Techniques</topic><topic>Escherichia coli - metabolism</topic><topic>Gene Expression Regulation - genetics</topic><topic>Metabolic Engineering - methods</topic><topic>Shikimic Acid - analogs & derivatives</topic><topic>Transcription Factors - metabolism</topic><topic>Transcriptome - genetics</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Li, Liangpo</creatorcontrib><creatorcontrib>Tu, Ran</creatorcontrib><creatorcontrib>Song, Guotian</creatorcontrib><creatorcontrib>Cheng, Jie</creatorcontrib><creatorcontrib>Chen, Wujiu</creatorcontrib><creatorcontrib>Li, Lin</creatorcontrib><creatorcontrib>Wang, Lixian</creatorcontrib><creatorcontrib>Wang, Qinhong</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>ACS synthetic biology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Li, Liangpo</au><au>Tu, Ran</au><au>Song, Guotian</au><au>Cheng, Jie</au><au>Chen, Wujiu</au><au>Li, Lin</au><au>Wang, Lixian</au><au>Wang, Qinhong</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Development of a Synthetic 3‑Dehydroshikimate Biosensor in Escherichia coli for Metabolite Monitoring and Genetic Screening</atitle><jtitle>ACS synthetic biology</jtitle><addtitle>ACS Synth. Biol</addtitle><date>2019-02-15</date><risdate>2019</risdate><volume>8</volume><issue>2</issue><spage>297</spage><epage>306</epage><pages>297-306</pages><issn>2161-5063</issn><eissn>2161-5063</eissn><abstract>Biosensors for target metabolites provide powerful high-throughput screening tools to obtain high-performing strains. However, well-characterized metabolite-sensing modules are often unavailable and limit rapid access to the robust biosensors with successful applications. In this study, we developed a strategy of transcriptome-assisted metabolite-sensing (TAMES) to identify the target metabolite-sensing module based on selectively comparative transcriptome analysis between the target metabolite producing and nonproducing strains and a subsequent quantative reverse transcription (RT-qPCR) evaluation. The strategy was applied to identify the sensing module cusR that responds positively to the metabolite 3-dehydroshikimate (DHS) and proved it was effective to narrow down the candidates. We further constructed the cusR-based synthetic biosensor and established the DHS biosensor-based high-throughput screening (HTS) platform to screen higher DHS-producing strains and successfully increased DHS production by more than 90%. This study demonstrated that the TAMES strategy was effective at exploiting the metabolite-sensing transcriptional regulator, and this could likely be extended to develop the biosensor-based HTS platforms for other molecules.</abstract><cop>United States</cop><pub>American Chemical Society</pub><pmid>30609888</pmid><doi>10.1021/acssynbio.8b00317</doi><tpages>10</tpages><orcidid>https://orcid.org/0000-0003-0004-2721</orcidid></addata></record> |
fulltext | fulltext |
identifier | ISSN: 2161-5063 |
ispartof | ACS synthetic biology, 2019-02, Vol.8 (2), p.297-306 |
issn | 2161-5063 2161-5063 |
language | eng |
recordid | cdi_proquest_miscellaneous_2164102721 |
source | MEDLINE; American Chemical Society Journals |
subjects | Biosensing Techniques Escherichia coli - metabolism Gene Expression Regulation - genetics Metabolic Engineering - methods Shikimic Acid - analogs & derivatives Transcription Factors - metabolism Transcriptome - genetics |
title | Development of a Synthetic 3‑Dehydroshikimate Biosensor in Escherichia coli for Metabolite Monitoring and Genetic Screening |
url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-23T09%3A35%3A24IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Development%20of%20a%20Synthetic%203%E2%80%91Dehydroshikimate%20Biosensor%20in%20Escherichia%20coli%20for%20Metabolite%20Monitoring%20and%20Genetic%20Screening&rft.jtitle=ACS%20synthetic%20biology&rft.au=Li,%20Liangpo&rft.date=2019-02-15&rft.volume=8&rft.issue=2&rft.spage=297&rft.epage=306&rft.pages=297-306&rft.issn=2161-5063&rft.eissn=2161-5063&rft_id=info:doi/10.1021/acssynbio.8b00317&rft_dat=%3Cproquest_cross%3E2164102721%3C/proquest_cross%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=2164102721&rft_id=info:pmid/30609888&rfr_iscdi=true |