A novel, genetically encoded whole-cell biosensor for directed evolution of myrcene synthase in Escherichia coli
β-myrcene is a high-value acyclic monoterpene. The low activity of myrcene synthase resulted to low biosynthetic titer of it. Biosensor is a promising tool applied for enzyme directed evolution. In this work, a novel genetically encoded biosensor responding to myrcene was established based on the My...
Gespeichert in:
| Veröffentlicht in: | Biosensors & bioelectronics 2023-05, Vol.228, p.115176-115176, Article 115176 |
|---|---|
| Hauptverfasser: | , , , , , , , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 115176 |
|---|---|
| container_issue | |
| container_start_page | 115176 |
| container_title | Biosensors & bioelectronics |
| container_volume | 228 |
| creator | Chen, Chang Liu, Jiajia Yao, Ge Bao, Shaoheng Wan, Xiukun Wang, Fuli Wang, Kang Song, Tianyu Han, Penggang Liu, Tiangang Jiang, Hui |
| description | β-myrcene is a high-value acyclic monoterpene. The low activity of myrcene synthase resulted to low biosynthetic titer of it. Biosensor is a promising tool applied for enzyme directed evolution. In this work, a novel genetically encoded biosensor responding to myrcene was established based on the MyrR regulator from Pseudomonas sp. Through sensing promoter characterization and engineering, the biosensor exhibiting excellent specificity and dynamic range was developed, and applied for directed evolution of myrcene synthase. After high-throughput screening of the myrcene synthase random mutation library, the best mutant R89G/N152S/D517N was obtained. Its catalytic efficiency was 1.47-fold than that of parent. Based on the mutants, the final production of myrcene reached 510.38 mg/L, which is the highest myrcene titer reported to date. This work demonstrates the great potential of whole-cell biosensor for improving enzymatic activity and the production of target metabolite. |
| doi_str_mv | 10.1016/j.bios.2023.115176 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_2786811816</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><els_id>S0956566323001185</els_id><sourcerecordid>2786811816</sourcerecordid><originalsourceid>FETCH-LOGICAL-c356t-b6d8aa5225890614bbd25184f262dce8e175e5c75ac514d9d1f938d8faad77253</originalsourceid><addsrcrecordid>eNp9kLFu2zAQhokiReOkfYEMAccMlcsjRYoCsgRBmhYI0KWdCYo81TRo0SFlB377SlCaMcPhlu_-w_8RcgVsDQzUt-26C6msOeNiDSChUR_ICnQjqpoLeUZWrJWqkkqJc3JRypYx1kDLPpFzoVoQWtcrsr-jQzpi_Er_4oBjcDbGE8XBJY-evmxSxMphjHR-hUNJmfbT-JDRjROBxxQPY0gDTT3dnbKbUmg5DePGFqRhoA_FbTAHtwmWuhTDZ_Kxt7Hgl9d9Sf58f_h9_6N6-vX48_7uqXJCqrHqlNfWSs6lbpmCuus8l6DrnivuHWqERqJ0jbROQu1bD30rtNe9tb5puBSX5GbJ3ef0fMAyml0ocxM7YDoUwxutNIAGNaF8QV1OpWTszT6Hnc0nA8zMps3WzPXNbNospqej69f8Q7dD_3byX-0E3C4ATi2PAbMpLkxicXFnfArv5f8DCcuQuA</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>2786811816</pqid></control><display><type>article</type><title>A novel, genetically encoded whole-cell biosensor for directed evolution of myrcene synthase in Escherichia coli</title><source>MEDLINE</source><source>Elsevier ScienceDirect Journals Complete</source><creator>Chen, Chang ; Liu, Jiajia ; Yao, Ge ; Bao, Shaoheng ; Wan, Xiukun ; Wang, Fuli ; Wang, Kang ; Song, Tianyu ; Han, Penggang ; Liu, Tiangang ; Jiang, Hui</creator><creatorcontrib>Chen, Chang ; Liu, Jiajia ; Yao, Ge ; Bao, Shaoheng ; Wan, Xiukun ; Wang, Fuli ; Wang, Kang ; Song, Tianyu ; Han, Penggang ; Liu, Tiangang ; Jiang, Hui</creatorcontrib><description>β-myrcene is a high-value acyclic monoterpene. The low activity of myrcene synthase resulted to low biosynthetic titer of it. Biosensor is a promising tool applied for enzyme directed evolution. In this work, a novel genetically encoded biosensor responding to myrcene was established based on the MyrR regulator from Pseudomonas sp. Through sensing promoter characterization and engineering, the biosensor exhibiting excellent specificity and dynamic range was developed, and applied for directed evolution of myrcene synthase. After high-throughput screening of the myrcene synthase random mutation library, the best mutant R89G/N152S/D517N was obtained. Its catalytic efficiency was 1.47-fold than that of parent. Based on the mutants, the final production of myrcene reached 510.38 mg/L, which is the highest myrcene titer reported to date. This work demonstrates the great potential of whole-cell biosensor for improving enzymatic activity and the production of target metabolite.</description><identifier>ISSN: 0956-5663</identifier><identifier>EISSN: 1873-4235</identifier><identifier>DOI: 10.1016/j.bios.2023.115176</identifier><identifier>PMID: 36913884</identifier><language>eng</language><publisher>England: Elsevier B.V</publisher><subject>Acyclic Monoterpenes - metabolism ; Biosensing Techniques ; Biosensor ; Directed evolution ; Escherichia coli - genetics ; Escherichia coli - metabolism ; High-throughput screening ; Monoterpenes - metabolism ; Myrcene</subject><ispartof>Biosensors & bioelectronics, 2023-05, Vol.228, p.115176-115176, Article 115176</ispartof><rights>2023</rights><rights>Copyright © 2023. Published by Elsevier B.V.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c356t-b6d8aa5225890614bbd25184f262dce8e175e5c75ac514d9d1f938d8faad77253</citedby><cites>FETCH-LOGICAL-c356t-b6d8aa5225890614bbd25184f262dce8e175e5c75ac514d9d1f938d8faad77253</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.bios.2023.115176$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>315,781,785,3551,27929,27930,46000</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/36913884$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Chen, Chang</creatorcontrib><creatorcontrib>Liu, Jiajia</creatorcontrib><creatorcontrib>Yao, Ge</creatorcontrib><creatorcontrib>Bao, Shaoheng</creatorcontrib><creatorcontrib>Wan, Xiukun</creatorcontrib><creatorcontrib>Wang, Fuli</creatorcontrib><creatorcontrib>Wang, Kang</creatorcontrib><creatorcontrib>Song, Tianyu</creatorcontrib><creatorcontrib>Han, Penggang</creatorcontrib><creatorcontrib>Liu, Tiangang</creatorcontrib><creatorcontrib>Jiang, Hui</creatorcontrib><title>A novel, genetically encoded whole-cell biosensor for directed evolution of myrcene synthase in Escherichia coli</title><title>Biosensors & bioelectronics</title><addtitle>Biosens Bioelectron</addtitle><description>β-myrcene is a high-value acyclic monoterpene. The low activity of myrcene synthase resulted to low biosynthetic titer of it. Biosensor is a promising tool applied for enzyme directed evolution. In this work, a novel genetically encoded biosensor responding to myrcene was established based on the MyrR regulator from Pseudomonas sp. Through sensing promoter characterization and engineering, the biosensor exhibiting excellent specificity and dynamic range was developed, and applied for directed evolution of myrcene synthase. After high-throughput screening of the myrcene synthase random mutation library, the best mutant R89G/N152S/D517N was obtained. Its catalytic efficiency was 1.47-fold than that of parent. Based on the mutants, the final production of myrcene reached 510.38 mg/L, which is the highest myrcene titer reported to date. This work demonstrates the great potential of whole-cell biosensor for improving enzymatic activity and the production of target metabolite.</description><subject>Acyclic Monoterpenes - metabolism</subject><subject>Biosensing Techniques</subject><subject>Biosensor</subject><subject>Directed evolution</subject><subject>Escherichia coli - genetics</subject><subject>Escherichia coli - metabolism</subject><subject>High-throughput screening</subject><subject>Monoterpenes - metabolism</subject><subject>Myrcene</subject><issn>0956-5663</issn><issn>1873-4235</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2023</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kLFu2zAQhokiReOkfYEMAccMlcsjRYoCsgRBmhYI0KWdCYo81TRo0SFlB377SlCaMcPhlu_-w_8RcgVsDQzUt-26C6msOeNiDSChUR_ICnQjqpoLeUZWrJWqkkqJc3JRypYx1kDLPpFzoVoQWtcrsr-jQzpi_Er_4oBjcDbGE8XBJY-evmxSxMphjHR-hUNJmfbT-JDRjROBxxQPY0gDTT3dnbKbUmg5DePGFqRhoA_FbTAHtwmWuhTDZ_Kxt7Hgl9d9Sf58f_h9_6N6-vX48_7uqXJCqrHqlNfWSs6lbpmCuus8l6DrnivuHWqERqJ0jbROQu1bD30rtNe9tb5puBSX5GbJ3ef0fMAyml0ocxM7YDoUwxutNIAGNaF8QV1OpWTszT6Hnc0nA8zMps3WzPXNbNospqej69f8Q7dD_3byX-0E3C4ATi2PAbMpLkxicXFnfArv5f8DCcuQuA</recordid><startdate>20230515</startdate><enddate>20230515</enddate><creator>Chen, Chang</creator><creator>Liu, Jiajia</creator><creator>Yao, Ge</creator><creator>Bao, Shaoheng</creator><creator>Wan, Xiukun</creator><creator>Wang, Fuli</creator><creator>Wang, Kang</creator><creator>Song, Tianyu</creator><creator>Han, Penggang</creator><creator>Liu, Tiangang</creator><creator>Jiang, Hui</creator><general>Elsevier B.V</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></search><sort><creationdate>20230515</creationdate><title>A novel, genetically encoded whole-cell biosensor for directed evolution of myrcene synthase in Escherichia coli</title><author>Chen, Chang ; Liu, Jiajia ; Yao, Ge ; Bao, Shaoheng ; Wan, Xiukun ; Wang, Fuli ; Wang, Kang ; Song, Tianyu ; Han, Penggang ; Liu, Tiangang ; Jiang, Hui</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c356t-b6d8aa5225890614bbd25184f262dce8e175e5c75ac514d9d1f938d8faad77253</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2023</creationdate><topic>Acyclic Monoterpenes - metabolism</topic><topic>Biosensing Techniques</topic><topic>Biosensor</topic><topic>Directed evolution</topic><topic>Escherichia coli - genetics</topic><topic>Escherichia coli - metabolism</topic><topic>High-throughput screening</topic><topic>Monoterpenes - metabolism</topic><topic>Myrcene</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Chen, Chang</creatorcontrib><creatorcontrib>Liu, Jiajia</creatorcontrib><creatorcontrib>Yao, Ge</creatorcontrib><creatorcontrib>Bao, Shaoheng</creatorcontrib><creatorcontrib>Wan, Xiukun</creatorcontrib><creatorcontrib>Wang, Fuli</creatorcontrib><creatorcontrib>Wang, Kang</creatorcontrib><creatorcontrib>Song, Tianyu</creatorcontrib><creatorcontrib>Han, Penggang</creatorcontrib><creatorcontrib>Liu, Tiangang</creatorcontrib><creatorcontrib>Jiang, Hui</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>Biosensors & bioelectronics</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Chen, Chang</au><au>Liu, Jiajia</au><au>Yao, Ge</au><au>Bao, Shaoheng</au><au>Wan, Xiukun</au><au>Wang, Fuli</au><au>Wang, Kang</au><au>Song, Tianyu</au><au>Han, Penggang</au><au>Liu, Tiangang</au><au>Jiang, Hui</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A novel, genetically encoded whole-cell biosensor for directed evolution of myrcene synthase in Escherichia coli</atitle><jtitle>Biosensors & bioelectronics</jtitle><addtitle>Biosens Bioelectron</addtitle><date>2023-05-15</date><risdate>2023</risdate><volume>228</volume><spage>115176</spage><epage>115176</epage><pages>115176-115176</pages><artnum>115176</artnum><issn>0956-5663</issn><eissn>1873-4235</eissn><abstract>β-myrcene is a high-value acyclic monoterpene. The low activity of myrcene synthase resulted to low biosynthetic titer of it. Biosensor is a promising tool applied for enzyme directed evolution. In this work, a novel genetically encoded biosensor responding to myrcene was established based on the MyrR regulator from Pseudomonas sp. Through sensing promoter characterization and engineering, the biosensor exhibiting excellent specificity and dynamic range was developed, and applied for directed evolution of myrcene synthase. After high-throughput screening of the myrcene synthase random mutation library, the best mutant R89G/N152S/D517N was obtained. Its catalytic efficiency was 1.47-fold than that of parent. Based on the mutants, the final production of myrcene reached 510.38 mg/L, which is the highest myrcene titer reported to date. This work demonstrates the great potential of whole-cell biosensor for improving enzymatic activity and the production of target metabolite.</abstract><cop>England</cop><pub>Elsevier B.V</pub><pmid>36913884</pmid><doi>10.1016/j.bios.2023.115176</doi><tpages>1</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0956-5663 |
| ispartof | Biosensors & bioelectronics, 2023-05, Vol.228, p.115176-115176, Article 115176 |
| issn | 0956-5663 1873-4235 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_2786811816 |
| source | MEDLINE; Elsevier ScienceDirect Journals Complete |
| subjects | Acyclic Monoterpenes - metabolism Biosensing Techniques Biosensor Directed evolution Escherichia coli - genetics Escherichia coli - metabolism High-throughput screening Monoterpenes - metabolism Myrcene |
| title | A novel, genetically encoded whole-cell biosensor for directed evolution of myrcene synthase in Escherichia coli |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-13T04%3A09%3A53IST&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=A%20novel,%20genetically%20encoded%20whole-cell%20biosensor%20for%20directed%20evolution%20of%20myrcene%20synthase%20in%20Escherichia%20coli&rft.jtitle=Biosensors%20&%20bioelectronics&rft.au=Chen,%20Chang&rft.date=2023-05-15&rft.volume=228&rft.spage=115176&rft.epage=115176&rft.pages=115176-115176&rft.artnum=115176&rft.issn=0956-5663&rft.eissn=1873-4235&rft_id=info:doi/10.1016/j.bios.2023.115176&rft_dat=%3Cproquest_cross%3E2786811816%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=2786811816&rft_id=info:pmid/36913884&rft_els_id=S0956566323001185&rfr_iscdi=true |