Systematic Optimization of Limonene Production in Engineered Escherichia coli
Limonene, a cyclic monoterpene, is widely used in food and cosmetics industries as well as in agriculture. In the work described herein, employing a systematic optimization strategy, we constructed an efficient platform for producing limonene via the heterologous mevalonate pathway in Escherichia co...
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Veröffentlicht in: | Journal of agricultural and food chemistry 2019-06, Vol.67 (25), p.7087-7097 |
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creator | Wu, Jihua Cheng, Si Cao, Jiayu Qiao, Jianjun Zhao, Guang-Rong |
description | Limonene, a cyclic monoterpene, is widely used in food and cosmetics industries as well as in agriculture. In the work described herein, employing a systematic optimization strategy, we constructed an efficient platform for producing limonene via the heterologous mevalonate pathway in Escherichia coli. By site-directed mutation of EfMvaS and tuning the initial translation of EfMvaE and EfMvaSA110G through ribosome binding site engineering, the upstream module for overproducing mevalonate was obtained. Expression of MmMK with ScPMK, ScPMD, and ScIDI under FAB80 promoter resulted in an efficient midstream module to produce 181.73 mg/L of limonene. Subsequently, coexpression of SlNPPS and MsLS in the downstream module led to a great improvement of limonene production to 694.61 mg/L. Finally, metabolically engineered strain ELIM78 produced 1.29 g/L of limonene in 84 h by fed-batch fermentation in a shake-flask. This is the first report on limonene biosynthesis in E. coli using neryl pyrophosphate synthase, which has promising potential for producing other monoterpenes. |
doi_str_mv | 10.1021/acs.jafc.9b01427 |
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In the work described herein, employing a systematic optimization strategy, we constructed an efficient platform for producing limonene via the heterologous mevalonate pathway in Escherichia coli. By site-directed mutation of EfMvaS and tuning the initial translation of EfMvaE and EfMvaSA110G through ribosome binding site engineering, the upstream module for overproducing mevalonate was obtained. Expression of MmMK with ScPMK, ScPMD, and ScIDI under FAB80 promoter resulted in an efficient midstream module to produce 181.73 mg/L of limonene. Subsequently, coexpression of SlNPPS and MsLS in the downstream module led to a great improvement of limonene production to 694.61 mg/L. Finally, metabolically engineered strain ELIM78 produced 1.29 g/L of limonene in 84 h by fed-batch fermentation in a shake-flask. This is the first report on limonene biosynthesis in E. coli using neryl pyrophosphate synthase, which has promising potential for producing other monoterpenes.</description><identifier>ISSN: 0021-8561</identifier><identifier>EISSN: 1520-5118</identifier><identifier>DOI: 10.1021/acs.jafc.9b01427</identifier><identifier>PMID: 31199132</identifier><language>eng</language><publisher>United States: American Chemical Society</publisher><subject>Batch Cell Culture Techniques ; Escherichia coli - genetics ; Escherichia coli - metabolism ; Fermentation ; Limonene - metabolism ; Metabolic Engineering ; Mevalonic Acid - metabolism ; Monoterpenes - metabolism</subject><ispartof>Journal of agricultural and food chemistry, 2019-06, Vol.67 (25), p.7087-7097</ispartof><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-a336t-a5e2eff2aa22c9ef902d04ee39fe388fc40069843bcb79e94006eff63c0c1b373</citedby><cites>FETCH-LOGICAL-a336t-a5e2eff2aa22c9ef902d04ee39fe388fc40069843bcb79e94006eff63c0c1b373</cites><orcidid>0000-0001-7626-5812 ; 0000-0002-6881-9042 ; 0000-0002-1548-9502</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/acs.jafc.9b01427$$EPDF$$P50$$Gacs$$H</linktopdf><linktohtml>$$Uhttps://pubs.acs.org/doi/10.1021/acs.jafc.9b01427$$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/31199132$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Wu, Jihua</creatorcontrib><creatorcontrib>Cheng, Si</creatorcontrib><creatorcontrib>Cao, Jiayu</creatorcontrib><creatorcontrib>Qiao, Jianjun</creatorcontrib><creatorcontrib>Zhao, Guang-Rong</creatorcontrib><title>Systematic Optimization of Limonene Production in Engineered Escherichia coli</title><title>Journal of agricultural and food chemistry</title><addtitle>J. Agric. Food Chem</addtitle><description>Limonene, a cyclic monoterpene, is widely used in food and cosmetics industries as well as in agriculture. In the work described herein, employing a systematic optimization strategy, we constructed an efficient platform for producing limonene via the heterologous mevalonate pathway in Escherichia coli. By site-directed mutation of EfMvaS and tuning the initial translation of EfMvaE and EfMvaSA110G through ribosome binding site engineering, the upstream module for overproducing mevalonate was obtained. Expression of MmMK with ScPMK, ScPMD, and ScIDI under FAB80 promoter resulted in an efficient midstream module to produce 181.73 mg/L of limonene. Subsequently, coexpression of SlNPPS and MsLS in the downstream module led to a great improvement of limonene production to 694.61 mg/L. Finally, metabolically engineered strain ELIM78 produced 1.29 g/L of limonene in 84 h by fed-batch fermentation in a shake-flask. This is the first report on limonene biosynthesis in E. coli using neryl pyrophosphate synthase, which has promising potential for producing other monoterpenes.</description><subject>Batch Cell Culture Techniques</subject><subject>Escherichia coli - genetics</subject><subject>Escherichia coli - metabolism</subject><subject>Fermentation</subject><subject>Limonene - metabolism</subject><subject>Metabolic Engineering</subject><subject>Mevalonic Acid - metabolism</subject><subject>Monoterpenes - metabolism</subject><issn>0021-8561</issn><issn>1520-5118</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2019</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp1kDtPwzAURi0EoqWwM6GMDKT4kZdHhMpDKioSMFuOc01dNXaxk6H8epy2sDFdX-t8n3QPQpcETwmm5FaqMF1Jraa8xiSj5REak5ziNCekOkZjHJm0ygsyQmchrDDGVV7iUzRihHBOGB2jl7dt6KCVnVHJYtOZ1nzHt7OJ08nctM6CheTVu6ZXu29jk5n9NBbAQ5PMglqCN2ppZKLc2pyjEy3XAS4Oc4I-Hmbv90_pfPH4fH83TyVjRZfKHChoTaWkVHHQHNMGZwCMa2BVpVWGccGrjNWqLjnwYY18wRRWpGYlm6Drfe_Gu68eQidaExSs19KC64OgLEZYxcmA4j2qvAvBgxYbb1rpt4JgMUgUUaIYJIqDxBi5OrT3dQvNX-DXWgRu9sAu6npv47H_9_0AryZ-VQ</recordid><startdate>20190626</startdate><enddate>20190626</enddate><creator>Wu, Jihua</creator><creator>Cheng, Si</creator><creator>Cao, Jiayu</creator><creator>Qiao, Jianjun</creator><creator>Zhao, Guang-Rong</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/0000-0001-7626-5812</orcidid><orcidid>https://orcid.org/0000-0002-6881-9042</orcidid><orcidid>https://orcid.org/0000-0002-1548-9502</orcidid></search><sort><creationdate>20190626</creationdate><title>Systematic Optimization of Limonene Production in Engineered Escherichia coli</title><author>Wu, Jihua ; Cheng, Si ; Cao, Jiayu ; Qiao, Jianjun ; Zhao, Guang-Rong</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-a336t-a5e2eff2aa22c9ef902d04ee39fe388fc40069843bcb79e94006eff63c0c1b373</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2019</creationdate><topic>Batch Cell Culture Techniques</topic><topic>Escherichia coli - genetics</topic><topic>Escherichia coli - metabolism</topic><topic>Fermentation</topic><topic>Limonene - metabolism</topic><topic>Metabolic Engineering</topic><topic>Mevalonic Acid - metabolism</topic><topic>Monoterpenes - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Wu, Jihua</creatorcontrib><creatorcontrib>Cheng, Si</creatorcontrib><creatorcontrib>Cao, Jiayu</creatorcontrib><creatorcontrib>Qiao, Jianjun</creatorcontrib><creatorcontrib>Zhao, Guang-Rong</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>Journal of agricultural and food chemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Wu, Jihua</au><au>Cheng, Si</au><au>Cao, Jiayu</au><au>Qiao, Jianjun</au><au>Zhao, Guang-Rong</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Systematic Optimization of Limonene Production in Engineered Escherichia coli</atitle><jtitle>Journal of agricultural and food chemistry</jtitle><addtitle>J. Agric. Food Chem</addtitle><date>2019-06-26</date><risdate>2019</risdate><volume>67</volume><issue>25</issue><spage>7087</spage><epage>7097</epage><pages>7087-7097</pages><issn>0021-8561</issn><eissn>1520-5118</eissn><abstract>Limonene, a cyclic monoterpene, is widely used in food and cosmetics industries as well as in agriculture. In the work described herein, employing a systematic optimization strategy, we constructed an efficient platform for producing limonene via the heterologous mevalonate pathway in Escherichia coli. By site-directed mutation of EfMvaS and tuning the initial translation of EfMvaE and EfMvaSA110G through ribosome binding site engineering, the upstream module for overproducing mevalonate was obtained. Expression of MmMK with ScPMK, ScPMD, and ScIDI under FAB80 promoter resulted in an efficient midstream module to produce 181.73 mg/L of limonene. Subsequently, coexpression of SlNPPS and MsLS in the downstream module led to a great improvement of limonene production to 694.61 mg/L. Finally, metabolically engineered strain ELIM78 produced 1.29 g/L of limonene in 84 h by fed-batch fermentation in a shake-flask. This is the first report on limonene biosynthesis in E. coli using neryl pyrophosphate synthase, which has promising potential for producing other monoterpenes.</abstract><cop>United States</cop><pub>American Chemical Society</pub><pmid>31199132</pmid><doi>10.1021/acs.jafc.9b01427</doi><tpages>11</tpages><orcidid>https://orcid.org/0000-0001-7626-5812</orcidid><orcidid>https://orcid.org/0000-0002-6881-9042</orcidid><orcidid>https://orcid.org/0000-0002-1548-9502</orcidid></addata></record> |
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subjects | Batch Cell Culture Techniques Escherichia coli - genetics Escherichia coli - metabolism Fermentation Limonene - metabolism Metabolic Engineering Mevalonic Acid - metabolism Monoterpenes - metabolism |
title | Systematic Optimization of Limonene Production in Engineered Escherichia coli |
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