Biosynthesis of the oxygenated diterpene nezukol in the medicinal plant Isodon rubescens is catalyzed by a pair of diterpene synthases
Plants produce an immense diversity of natural products (i.e. secondary or specialized metabolites) that offer a rich source of known and potentially new pharmaceuticals and other desirable bioproducts. The Traditional Chinese Medicinal plant Isodon rubescens (Lamiaceae) contains an array of bioacti...
Gespeichert in:
| Veröffentlicht in: | PloS one 2017-04, Vol.12 (4), p.e0176507-e0176507 |
|---|---|
| Hauptverfasser: | , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | e0176507 |
|---|---|
| container_issue | 4 |
| container_start_page | e0176507 |
| container_title | PloS one |
| container_volume | 12 |
| creator | Pelot, Kyle A Hagelthorn, Lynne M Addison, J Bennett Zerbe, Philipp |
| description | Plants produce an immense diversity of natural products (i.e. secondary or specialized metabolites) that offer a rich source of known and potentially new pharmaceuticals and other desirable bioproducts. The Traditional Chinese Medicinal plant Isodon rubescens (Lamiaceae) contains an array of bioactive labdane-related diterpenoid natural products. Of these, the ent-kauranoid oridonin is the most prominent specialized metabolite that has been extensively studied for its potent antimicrobial and anticancer efficacy. Mining of a previously established transcriptome of I. rubescens leaf tissue identified seven diterpene synthase (diTPSs) candidates. Here we report the functional characterization of four I. rubescens diTPSs. IrTPS5 and IrTPS3 were identified as an ent-copalyl diphosphate (CPP) synthase and a (+)-CPP synthase, respectively. Distinct transcript abundance of IrTPS5 and the predicted ent-CPP synthase IrTPS1 suggested a role of IrTPS5 in specialized ent-kaurene metabolism possibly en route to oridonin. Nicotiana benthamiana co-expression assays demonstrated that IrTPS4 functions sequentially with IrTPS3 to form miltiradiene. In addition, IrTPS2 converted the IrTPS3 product (+)-CPP into the hydroxylated tricyclic diterpene nezukol not previously identified in I. rubescens. Metabolite profiling verified the presence of nezukol in I. rubescens leaf tissue. The proposed IrTPS2-catalyzed reaction mechanism proceeds via the common ionization of the diphosphate group of (+)-CPP, followed by formation of an intermediary pimar-15-en-8-yl+ carbocation and neutralization of the carbocation by water capture at C-8 to yield nezukol, as confirmed by nuclear magnetic resonance (NMR) analysis. Oxygenation activity is rare for the family of class I diTPSs and offers new catalysts for developing metabolic engineering platforms to produce a broader spectrum of bioactive diterpenoid natural products. |
| doi_str_mv | 10.1371/journal.pone.0176507 |
| format | Article |
| fullrecord | <record><control><sourceid>gale_plos_</sourceid><recordid>TN_cdi_plos_journals_1892318471</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><galeid>A493764981</galeid><doaj_id>oai_doaj_org_article_113dae66d0854ab088fad4e9a8ea1805</doaj_id><sourcerecordid>A493764981</sourcerecordid><originalsourceid>FETCH-LOGICAL-c692t-7c17ad620736fd10afbf3d343e91cb204b46784b6c1da889d3fb0973f8b187223</originalsourceid><addsrcrecordid>eNqNk12P1CAUhhujcdfRf2CUxMToxYxQKNAbk3XjxySbbOLXLTktdIaxA7VQs7M_wN8t87Hr1OyF4QICz3kP54WTZU8JnhEqyJuVH3oH7azzzswwEbzA4l52SkqaT3mO6f2j9Un2KIQVxgWVnD_MTnLJWFHk_DT7_c76sHFxaYINyDcorZC_2iyMg2g00jaavjPOIGeuhx--RdbtmLXRtrbpAqhrwUU0D157h_qhMqE2LqAkV0OEdnOdZKoNAtSB7bcp_mruMkMw4XH2oIE2mCeHeZJ9-_D-6_mn6cXlx_n52cW05mUep6ImAnSqSFDeaIKhqRqqKaOmJHWVY1YxLiSreE00SFlq2lS4FLSRFZEiz-kke77X7Vof1MHCoIgsc0okEyQR8z2hPaxU19s19Bvlwardhu8XCvpo69YoQqgGw7nGsmBQYSkb0MyUIA0QmcyeZG8P2YYq-ZVsiT20I9HxibNLtfC_VMFwUQqcBF4dBHr_czAhqrVN7rbJceOH_b1FXrCSJ_TFP-jd1R2oBaQCrGt8yltvRdUZK6ngrJRbanYHlYY2a1un_9bYtD8KeD0KSEw0V3EBQwhq_uXz_7OX38fsyyN2aaCNy-DbIVrvwhhke7DufQi9aW5NJlht2-XGDbVtF3VolxT27PiBboNu-oP-AapOEgg</addsrcrecordid><sourcetype>Open Website</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>1892318471</pqid></control><display><type>article</type><title>Biosynthesis of the oxygenated diterpene nezukol in the medicinal plant Isodon rubescens is catalyzed by a pair of diterpene synthases</title><source>MEDLINE</source><source>DOAJ Directory of Open Access Journals</source><source>Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals</source><source>PubMed Central</source><source>Free Full-Text Journals in Chemistry</source><source>Public Library of Science (PLoS)</source><creator>Pelot, Kyle A ; Hagelthorn, Lynne M ; Addison, J Bennett ; Zerbe, Philipp</creator><contributor>Imai, Ryozo</contributor><creatorcontrib>Pelot, Kyle A ; Hagelthorn, Lynne M ; Addison, J Bennett ; Zerbe, Philipp ; Imai, Ryozo</creatorcontrib><description>Plants produce an immense diversity of natural products (i.e. secondary or specialized metabolites) that offer a rich source of known and potentially new pharmaceuticals and other desirable bioproducts. The Traditional Chinese Medicinal plant Isodon rubescens (Lamiaceae) contains an array of bioactive labdane-related diterpenoid natural products. Of these, the ent-kauranoid oridonin is the most prominent specialized metabolite that has been extensively studied for its potent antimicrobial and anticancer efficacy. Mining of a previously established transcriptome of I. rubescens leaf tissue identified seven diterpene synthase (diTPSs) candidates. Here we report the functional characterization of four I. rubescens diTPSs. IrTPS5 and IrTPS3 were identified as an ent-copalyl diphosphate (CPP) synthase and a (+)-CPP synthase, respectively. Distinct transcript abundance of IrTPS5 and the predicted ent-CPP synthase IrTPS1 suggested a role of IrTPS5 in specialized ent-kaurene metabolism possibly en route to oridonin. Nicotiana benthamiana co-expression assays demonstrated that IrTPS4 functions sequentially with IrTPS3 to form miltiradiene. In addition, IrTPS2 converted the IrTPS3 product (+)-CPP into the hydroxylated tricyclic diterpene nezukol not previously identified in I. rubescens. Metabolite profiling verified the presence of nezukol in I. rubescens leaf tissue. The proposed IrTPS2-catalyzed reaction mechanism proceeds via the common ionization of the diphosphate group of (+)-CPP, followed by formation of an intermediary pimar-15-en-8-yl+ carbocation and neutralization of the carbocation by water capture at C-8 to yield nezukol, as confirmed by nuclear magnetic resonance (NMR) analysis. Oxygenation activity is rare for the family of class I diTPSs and offers new catalysts for developing metabolic engineering platforms to produce a broader spectrum of bioactive diterpenoid natural products.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0176507</identifier><identifier>PMID: 28445526</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Alkyl and Aryl Transferases - chemistry ; Alkyl and Aryl Transferases - classification ; Alkyl and Aryl Transferases - metabolism ; Amino acid sequence ; Analysis ; Angiosperms ; Anticancer properties ; Biocatalysis ; Biological products ; Biology and Life Sciences ; Biosynthesis ; Bushy stunt ; Cancer ; Catalysis ; Catalysts ; Cloning ; Cloning, Molecular ; Cyclic AMP ; Cytochrome ; Cytochrome P450 ; Deoxyribonucleic acid ; Diterpenes ; Diterpenes - chemistry ; Diterpenes - metabolism ; Diterpenes, Kaurane - biosynthesis ; Diterpenes, Kaurane - chemistry ; DNA ; DNA fingerprinting ; E coli ; Economic importance ; Engineering ; Enzymes ; Essential oils ; Foliage ; Forskolin ; Gas Chromatography-Mass Spectrometry ; Gene Expression ; Gene silencing ; Herbal medicine ; Ionization ; Isodon - chemistry ; Isodon - genetics ; Isodon - metabolism ; Lamiaceae ; Leaves ; Magnetic resonance ; Magnetic Resonance Spectroscopy ; Medicinal plants ; Medicine and Health Sciences ; Metabolism ; Metabolites ; Metabolome ; Mining ; Mutagenesis ; Natural products ; Neutralization ; Nicotiana - chemistry ; Nicotiana - metabolism ; NMR ; Nuclear magnetic resonance ; Oxygenation ; Phylogeny ; Physical Sciences ; Plant biology ; Plant Leaves - chemistry ; Plant Leaves - metabolism ; Plant Proteins - chemistry ; Plant Proteins - classification ; Plant Proteins - metabolism ; Plant tissues ; Plants ; Plants, Medicinal - chemistry ; Plants, Medicinal - metabolism ; Proteins ; Reaction mechanisms ; Residues ; Secondary metabolites ; Sesquiterpenes ; Species diversity ; Transcription ; Tumors</subject><ispartof>PloS one, 2017-04, Vol.12 (4), p.e0176507-e0176507</ispartof><rights>COPYRIGHT 2017 Public Library of Science</rights><rights>2017 Pelot et al. This is an open access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2017 Pelot et al 2017 Pelot et al</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c692t-7c17ad620736fd10afbf3d343e91cb204b46784b6c1da889d3fb0973f8b187223</citedby><cites>FETCH-LOGICAL-c692t-7c17ad620736fd10afbf3d343e91cb204b46784b6c1da889d3fb0973f8b187223</cites><orcidid>0000-0001-5163-9523</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC5405970/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC5405970/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,723,776,780,860,881,2095,2914,23846,27903,27904,53769,53771,79346,79347</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/28445526$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><contributor>Imai, Ryozo</contributor><creatorcontrib>Pelot, Kyle A</creatorcontrib><creatorcontrib>Hagelthorn, Lynne M</creatorcontrib><creatorcontrib>Addison, J Bennett</creatorcontrib><creatorcontrib>Zerbe, Philipp</creatorcontrib><title>Biosynthesis of the oxygenated diterpene nezukol in the medicinal plant Isodon rubescens is catalyzed by a pair of diterpene synthases</title><title>PloS one</title><addtitle>PLoS One</addtitle><description>Plants produce an immense diversity of natural products (i.e. secondary or specialized metabolites) that offer a rich source of known and potentially new pharmaceuticals and other desirable bioproducts. The Traditional Chinese Medicinal plant Isodon rubescens (Lamiaceae) contains an array of bioactive labdane-related diterpenoid natural products. Of these, the ent-kauranoid oridonin is the most prominent specialized metabolite that has been extensively studied for its potent antimicrobial and anticancer efficacy. Mining of a previously established transcriptome of I. rubescens leaf tissue identified seven diterpene synthase (diTPSs) candidates. Here we report the functional characterization of four I. rubescens diTPSs. IrTPS5 and IrTPS3 were identified as an ent-copalyl diphosphate (CPP) synthase and a (+)-CPP synthase, respectively. Distinct transcript abundance of IrTPS5 and the predicted ent-CPP synthase IrTPS1 suggested a role of IrTPS5 in specialized ent-kaurene metabolism possibly en route to oridonin. Nicotiana benthamiana co-expression assays demonstrated that IrTPS4 functions sequentially with IrTPS3 to form miltiradiene. In addition, IrTPS2 converted the IrTPS3 product (+)-CPP into the hydroxylated tricyclic diterpene nezukol not previously identified in I. rubescens. Metabolite profiling verified the presence of nezukol in I. rubescens leaf tissue. The proposed IrTPS2-catalyzed reaction mechanism proceeds via the common ionization of the diphosphate group of (+)-CPP, followed by formation of an intermediary pimar-15-en-8-yl+ carbocation and neutralization of the carbocation by water capture at C-8 to yield nezukol, as confirmed by nuclear magnetic resonance (NMR) analysis. Oxygenation activity is rare for the family of class I diTPSs and offers new catalysts for developing metabolic engineering platforms to produce a broader spectrum of bioactive diterpenoid natural products.</description><subject>Alkyl and Aryl Transferases - chemistry</subject><subject>Alkyl and Aryl Transferases - classification</subject><subject>Alkyl and Aryl Transferases - metabolism</subject><subject>Amino acid sequence</subject><subject>Analysis</subject><subject>Angiosperms</subject><subject>Anticancer properties</subject><subject>Biocatalysis</subject><subject>Biological products</subject><subject>Biology and Life Sciences</subject><subject>Biosynthesis</subject><subject>Bushy stunt</subject><subject>Cancer</subject><subject>Catalysis</subject><subject>Catalysts</subject><subject>Cloning</subject><subject>Cloning, Molecular</subject><subject>Cyclic AMP</subject><subject>Cytochrome</subject><subject>Cytochrome P450</subject><subject>Deoxyribonucleic acid</subject><subject>Diterpenes</subject><subject>Diterpenes - chemistry</subject><subject>Diterpenes - metabolism</subject><subject>Diterpenes, Kaurane - biosynthesis</subject><subject>Diterpenes, Kaurane - chemistry</subject><subject>DNA</subject><subject>DNA fingerprinting</subject><subject>E coli</subject><subject>Economic importance</subject><subject>Engineering</subject><subject>Enzymes</subject><subject>Essential oils</subject><subject>Foliage</subject><subject>Forskolin</subject><subject>Gas Chromatography-Mass Spectrometry</subject><subject>Gene Expression</subject><subject>Gene silencing</subject><subject>Herbal medicine</subject><subject>Ionization</subject><subject>Isodon - chemistry</subject><subject>Isodon - genetics</subject><subject>Isodon - metabolism</subject><subject>Lamiaceae</subject><subject>Leaves</subject><subject>Magnetic resonance</subject><subject>Magnetic Resonance Spectroscopy</subject><subject>Medicinal plants</subject><subject>Medicine and Health Sciences</subject><subject>Metabolism</subject><subject>Metabolites</subject><subject>Metabolome</subject><subject>Mining</subject><subject>Mutagenesis</subject><subject>Natural products</subject><subject>Neutralization</subject><subject>Nicotiana - chemistry</subject><subject>Nicotiana - metabolism</subject><subject>NMR</subject><subject>Nuclear magnetic resonance</subject><subject>Oxygenation</subject><subject>Phylogeny</subject><subject>Physical Sciences</subject><subject>Plant biology</subject><subject>Plant Leaves - chemistry</subject><subject>Plant Leaves - metabolism</subject><subject>Plant Proteins - chemistry</subject><subject>Plant Proteins - classification</subject><subject>Plant Proteins - metabolism</subject><subject>Plant tissues</subject><subject>Plants</subject><subject>Plants, Medicinal - chemistry</subject><subject>Plants, Medicinal - metabolism</subject><subject>Proteins</subject><subject>Reaction mechanisms</subject><subject>Residues</subject><subject>Secondary metabolites</subject><subject>Sesquiterpenes</subject><subject>Species diversity</subject><subject>Transcription</subject><subject>Tumors</subject><issn>1932-6203</issn><issn>1932-6203</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2017</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>BENPR</sourceid><sourceid>DOA</sourceid><recordid>eNqNk12P1CAUhhujcdfRf2CUxMToxYxQKNAbk3XjxySbbOLXLTktdIaxA7VQs7M_wN8t87Hr1OyF4QICz3kP54WTZU8JnhEqyJuVH3oH7azzzswwEbzA4l52SkqaT3mO6f2j9Un2KIQVxgWVnD_MTnLJWFHk_DT7_c76sHFxaYINyDcorZC_2iyMg2g00jaavjPOIGeuhx--RdbtmLXRtrbpAqhrwUU0D157h_qhMqE2LqAkV0OEdnOdZKoNAtSB7bcp_mruMkMw4XH2oIE2mCeHeZJ9-_D-6_mn6cXlx_n52cW05mUep6ImAnSqSFDeaIKhqRqqKaOmJHWVY1YxLiSreE00SFlq2lS4FLSRFZEiz-kke77X7Vof1MHCoIgsc0okEyQR8z2hPaxU19s19Bvlwardhu8XCvpo69YoQqgGw7nGsmBQYSkb0MyUIA0QmcyeZG8P2YYq-ZVsiT20I9HxibNLtfC_VMFwUQqcBF4dBHr_czAhqrVN7rbJceOH_b1FXrCSJ_TFP-jd1R2oBaQCrGt8yltvRdUZK6ngrJRbanYHlYY2a1un_9bYtD8KeD0KSEw0V3EBQwhq_uXz_7OX38fsyyN2aaCNy-DbIVrvwhhke7DufQi9aW5NJlht2-XGDbVtF3VolxT27PiBboNu-oP-AapOEgg</recordid><startdate>20170426</startdate><enddate>20170426</enddate><creator>Pelot, Kyle A</creator><creator>Hagelthorn, Lynne M</creator><creator>Addison, J Bennett</creator><creator>Zerbe, Philipp</creator><general>Public Library of Science</general><general>Public Library of Science (PLoS)</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>IOV</scope><scope>ISR</scope><scope>3V.</scope><scope>7QG</scope><scope>7QL</scope><scope>7QO</scope><scope>7RV</scope><scope>7SN</scope><scope>7SS</scope><scope>7T5</scope><scope>7TG</scope><scope>7TM</scope><scope>7U9</scope><scope>7X2</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8AO</scope><scope>8C1</scope><scope>8FD</scope><scope>8FE</scope><scope>8FG</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABJCF</scope><scope>ABUWG</scope><scope>AEUYN</scope><scope>AFKRA</scope><scope>ARAPS</scope><scope>ATCPS</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BGLVJ</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>D1I</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>KB.</scope><scope>KB0</scope><scope>KL.</scope><scope>L6V</scope><scope>LK8</scope><scope>M0K</scope><scope>M0S</scope><scope>M1P</scope><scope>M7N</scope><scope>M7P</scope><scope>M7S</scope><scope>NAPCQ</scope><scope>P5Z</scope><scope>P62</scope><scope>P64</scope><scope>PATMY</scope><scope>PDBOC</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>PTHSS</scope><scope>PYCSY</scope><scope>RC3</scope><scope>7X8</scope><scope>5PM</scope><scope>DOA</scope><orcidid>https://orcid.org/0000-0001-5163-9523</orcidid></search><sort><creationdate>20170426</creationdate><title>Biosynthesis of the oxygenated diterpene nezukol in the medicinal plant Isodon rubescens is catalyzed by a pair of diterpene synthases</title><author>Pelot, Kyle A ; Hagelthorn, Lynne M ; Addison, J Bennett ; Zerbe, Philipp</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c692t-7c17ad620736fd10afbf3d343e91cb204b46784b6c1da889d3fb0973f8b187223</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2017</creationdate><topic>Alkyl and Aryl Transferases - chemistry</topic><topic>Alkyl and Aryl Transferases - classification</topic><topic>Alkyl and Aryl Transferases - metabolism</topic><topic>Amino acid sequence</topic><topic>Analysis</topic><topic>Angiosperms</topic><topic>Anticancer properties</topic><topic>Biocatalysis</topic><topic>Biological products</topic><topic>Biology and Life Sciences</topic><topic>Biosynthesis</topic><topic>Bushy stunt</topic><topic>Cancer</topic><topic>Catalysis</topic><topic>Catalysts</topic><topic>Cloning</topic><topic>Cloning, Molecular</topic><topic>Cyclic AMP</topic><topic>Cytochrome</topic><topic>Cytochrome P450</topic><topic>Deoxyribonucleic acid</topic><topic>Diterpenes</topic><topic>Diterpenes - chemistry</topic><topic>Diterpenes - metabolism</topic><topic>Diterpenes, Kaurane - biosynthesis</topic><topic>Diterpenes, Kaurane - chemistry</topic><topic>DNA</topic><topic>DNA fingerprinting</topic><topic>E coli</topic><topic>Economic importance</topic><topic>Engineering</topic><topic>Enzymes</topic><topic>Essential oils</topic><topic>Foliage</topic><topic>Forskolin</topic><topic>Gas Chromatography-Mass Spectrometry</topic><topic>Gene Expression</topic><topic>Gene silencing</topic><topic>Herbal medicine</topic><topic>Ionization</topic><topic>Isodon - chemistry</topic><topic>Isodon - genetics</topic><topic>Isodon - metabolism</topic><topic>Lamiaceae</topic><topic>Leaves</topic><topic>Magnetic resonance</topic><topic>Magnetic Resonance Spectroscopy</topic><topic>Medicinal plants</topic><topic>Medicine and Health Sciences</topic><topic>Metabolism</topic><topic>Metabolites</topic><topic>Metabolome</topic><topic>Mining</topic><topic>Mutagenesis</topic><topic>Natural products</topic><topic>Neutralization</topic><topic>Nicotiana - chemistry</topic><topic>Nicotiana - metabolism</topic><topic>NMR</topic><topic>Nuclear magnetic resonance</topic><topic>Oxygenation</topic><topic>Phylogeny</topic><topic>Physical Sciences</topic><topic>Plant biology</topic><topic>Plant Leaves - chemistry</topic><topic>Plant Leaves - metabolism</topic><topic>Plant Proteins - chemistry</topic><topic>Plant Proteins - classification</topic><topic>Plant Proteins - metabolism</topic><topic>Plant tissues</topic><topic>Plants</topic><topic>Plants, Medicinal - chemistry</topic><topic>Plants, Medicinal - metabolism</topic><topic>Proteins</topic><topic>Reaction mechanisms</topic><topic>Residues</topic><topic>Secondary metabolites</topic><topic>Sesquiterpenes</topic><topic>Species diversity</topic><topic>Transcription</topic><topic>Tumors</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Pelot, Kyle A</creatorcontrib><creatorcontrib>Hagelthorn, Lynne M</creatorcontrib><creatorcontrib>Addison, J Bennett</creatorcontrib><creatorcontrib>Zerbe, Philipp</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Gale In Context: Opposing Viewpoints</collection><collection>Gale In Context: Science</collection><collection>ProQuest Central (Corporate)</collection><collection>Animal Behavior Abstracts</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Biotechnology Research Abstracts</collection><collection>Nursing & Allied Health Database</collection><collection>Ecology Abstracts</collection><collection>Entomology Abstracts (Full archive)</collection><collection>Immunology Abstracts</collection><collection>Meteorological & Geoastrophysical Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Agricultural Science Collection</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Public Health Database</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Technology Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>Materials Science & Engineering Collection</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest One Sustainability</collection><collection>ProQuest Central UK/Ireland</collection><collection>Advanced Technologies & Aerospace Collection</collection><collection>Agricultural & Environmental Science Collection</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Technology Collection</collection><collection>Natural Science Collection</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ProQuest One Community College</collection><collection>ProQuest Materials Science Collection</collection><collection>ProQuest Central Korea</collection><collection>Engineering Research Database</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Materials Science Database</collection><collection>Nursing & Allied Health Database (Alumni Edition)</collection><collection>Meteorological & Geoastrophysical Abstracts - Academic</collection><collection>ProQuest Engineering Collection</collection><collection>ProQuest Biological Science Collection</collection><collection>Agricultural Science Database</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biological Science Database</collection><collection>Engineering Database</collection><collection>Nursing & Allied Health Premium</collection><collection>Advanced Technologies & Aerospace Database</collection><collection>ProQuest Advanced Technologies & Aerospace Collection</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Environmental Science Database</collection><collection>Materials Science Collection</collection><collection>Publicly Available Content Database</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central China</collection><collection>Engineering Collection</collection><collection>Environmental Science Collection</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><collection>DOAJ Directory of Open Access Journals</collection><jtitle>PloS one</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Pelot, Kyle A</au><au>Hagelthorn, Lynne M</au><au>Addison, J Bennett</au><au>Zerbe, Philipp</au><au>Imai, Ryozo</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Biosynthesis of the oxygenated diterpene nezukol in the medicinal plant Isodon rubescens is catalyzed by a pair of diterpene synthases</atitle><jtitle>PloS one</jtitle><addtitle>PLoS One</addtitle><date>2017-04-26</date><risdate>2017</risdate><volume>12</volume><issue>4</issue><spage>e0176507</spage><epage>e0176507</epage><pages>e0176507-e0176507</pages><issn>1932-6203</issn><eissn>1932-6203</eissn><abstract>Plants produce an immense diversity of natural products (i.e. secondary or specialized metabolites) that offer a rich source of known and potentially new pharmaceuticals and other desirable bioproducts. The Traditional Chinese Medicinal plant Isodon rubescens (Lamiaceae) contains an array of bioactive labdane-related diterpenoid natural products. Of these, the ent-kauranoid oridonin is the most prominent specialized metabolite that has been extensively studied for its potent antimicrobial and anticancer efficacy. Mining of a previously established transcriptome of I. rubescens leaf tissue identified seven diterpene synthase (diTPSs) candidates. Here we report the functional characterization of four I. rubescens diTPSs. IrTPS5 and IrTPS3 were identified as an ent-copalyl diphosphate (CPP) synthase and a (+)-CPP synthase, respectively. Distinct transcript abundance of IrTPS5 and the predicted ent-CPP synthase IrTPS1 suggested a role of IrTPS5 in specialized ent-kaurene metabolism possibly en route to oridonin. Nicotiana benthamiana co-expression assays demonstrated that IrTPS4 functions sequentially with IrTPS3 to form miltiradiene. In addition, IrTPS2 converted the IrTPS3 product (+)-CPP into the hydroxylated tricyclic diterpene nezukol not previously identified in I. rubescens. Metabolite profiling verified the presence of nezukol in I. rubescens leaf tissue. The proposed IrTPS2-catalyzed reaction mechanism proceeds via the common ionization of the diphosphate group of (+)-CPP, followed by formation of an intermediary pimar-15-en-8-yl+ carbocation and neutralization of the carbocation by water capture at C-8 to yield nezukol, as confirmed by nuclear magnetic resonance (NMR) analysis. Oxygenation activity is rare for the family of class I diTPSs and offers new catalysts for developing metabolic engineering platforms to produce a broader spectrum of bioactive diterpenoid natural products.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>28445526</pmid><doi>10.1371/journal.pone.0176507</doi><tpages>e0176507</tpages><orcidid>https://orcid.org/0000-0001-5163-9523</orcidid><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1932-6203 |
| ispartof | PloS one, 2017-04, Vol.12 (4), p.e0176507-e0176507 |
| issn | 1932-6203 1932-6203 |
| language | eng |
| recordid | cdi_plos_journals_1892318471 |
| source | MEDLINE; DOAJ Directory of Open Access Journals; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; PubMed Central; Free Full-Text Journals in Chemistry; Public Library of Science (PLoS) |
| subjects | Alkyl and Aryl Transferases - chemistry Alkyl and Aryl Transferases - classification Alkyl and Aryl Transferases - metabolism Amino acid sequence Analysis Angiosperms Anticancer properties Biocatalysis Biological products Biology and Life Sciences Biosynthesis Bushy stunt Cancer Catalysis Catalysts Cloning Cloning, Molecular Cyclic AMP Cytochrome Cytochrome P450 Deoxyribonucleic acid Diterpenes Diterpenes - chemistry Diterpenes - metabolism Diterpenes, Kaurane - biosynthesis Diterpenes, Kaurane - chemistry DNA DNA fingerprinting E coli Economic importance Engineering Enzymes Essential oils Foliage Forskolin Gas Chromatography-Mass Spectrometry Gene Expression Gene silencing Herbal medicine Ionization Isodon - chemistry Isodon - genetics Isodon - metabolism Lamiaceae Leaves Magnetic resonance Magnetic Resonance Spectroscopy Medicinal plants Medicine and Health Sciences Metabolism Metabolites Metabolome Mining Mutagenesis Natural products Neutralization Nicotiana - chemistry Nicotiana - metabolism NMR Nuclear magnetic resonance Oxygenation Phylogeny Physical Sciences Plant biology Plant Leaves - chemistry Plant Leaves - metabolism Plant Proteins - chemistry Plant Proteins - classification Plant Proteins - metabolism Plant tissues Plants Plants, Medicinal - chemistry Plants, Medicinal - metabolism Proteins Reaction mechanisms Residues Secondary metabolites Sesquiterpenes Species diversity Transcription Tumors |
| title | Biosynthesis of the oxygenated diterpene nezukol in the medicinal plant Isodon rubescens is catalyzed by a pair of diterpene synthases |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-27T00%3A45%3A16IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-gale_plos_&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Biosynthesis%20of%20the%20oxygenated%20diterpene%20nezukol%20in%20the%20medicinal%20plant%20Isodon%20rubescens%20is%20catalyzed%20by%20a%20pair%20of%20diterpene%20synthases&rft.jtitle=PloS%20one&rft.au=Pelot,%20Kyle%20A&rft.date=2017-04-26&rft.volume=12&rft.issue=4&rft.spage=e0176507&rft.epage=e0176507&rft.pages=e0176507-e0176507&rft.issn=1932-6203&rft.eissn=1932-6203&rft_id=info:doi/10.1371/journal.pone.0176507&rft_dat=%3Cgale_plos_%3EA493764981%3C/gale_plos_%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=1892318471&rft_id=info:pmid/28445526&rft_galeid=A493764981&rft_doaj_id=oai_doaj_org_article_113dae66d0854ab088fad4e9a8ea1805&rfr_iscdi=true |