Nepeta cataria L. (catnip) can serve as a chassis for the engineering of secondary metabolic pathways
Objective Evaluation of Nepeta cataria as a host with specific endogenous metabolite background for transient expression and metabolic engineering of secondary biosynthetic sequences. Results The reporter gene gfp::licBM3 as well as three biosynthetic genes leading to the formation of the cannabinoi...
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| Veröffentlicht in: | Biotechnology letters 2024-10, Vol.46 (5), p.843-850 |
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| creator | Geissler, Marcus Neubauer, Christoph Sheludko, Yuriy V. Brückner, Adrian Warzecha, Heribert |
| description | Objective
Evaluation of
Nepeta cataria
as a host with specific endogenous metabolite background for transient expression and metabolic engineering of secondary biosynthetic sequences.
Results
The reporter gene
gfp::licBM3
as well as three biosynthetic genes leading to the formation of the cannabinoid precursor olivetolic acid were adopted to the modular cloning standard GoldenBraid, transiently expressed in two chemotypes of
N. cataria
and compared to
Nicotiana benthamiana
. To estimate the expression efficiency in both hosts, quantification of the reporter activity was carried out with a sensitive and specific lichenase assay. While
N.
benthamiana
exhibited lichenase activity of 676 ± 94 μmol g
−1
s
−1
,
N. cataria
cultivar ‘1000’, and the cultivar ‘Citriodora’ showed an activity of 37 ± 8 μmol g
−1
s
−1
and 18 ± 4 μmol g
−1
s
−1
, respectively. Further, combinatorial expression of genes involved in cannabinoid biosynthetic pathway
acyl-activating enzyme 1
(
aae1
),
olivetol synthase
(
ols
) and
olivetolic acid cyclase
(
oac
) in
N. cataria
cv. resulted presumably in the in vivo production of olivetolic acid glycosides.
Conclusion
Nepeta cataria
is amenable to
Agrobacterium
-mediated transient expression and could serve as a novel chassis for the engineering of secondary metabolic pathways and transient evaluation of heterologous genes. |
| doi_str_mv | 10.1007/s10529-024-03489-w |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_pubme</sourceid><recordid>TN_cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_11415451</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>3052594986</sourcerecordid><originalsourceid>FETCH-LOGICAL-c426t-26d42fdbbe6040242ecdaa50943bddc5c33b2b4421e5921be6106b23d7be58183</originalsourceid><addsrcrecordid>eNp9kctuFDEQRS0EIpPAD7BAltiERQe_3V4hFEFAGsEG1pbtrp5x1GM3dk9G-XscJoTHgpUt-9StunURekHJBSVEv6mUSGY6wkRHuOhNd3iEVlRq3imt1WO0IlTQTgrDTtBprdeEEKOJfopOeK-pVoqtEHyGGRaHg1tciQ6vL_B5u6c4v25vCVcoN4BdxQ3ZulpjxWMueNkChrSJCaDEtMF5bGTIaXDlFu-aoM9TDHh2y_bgbusz9GR0U4Xn9-cZ-vbh_dfLj936y9Wny3frLgimlo6pQbBx8B4UEc0WgzA4J4kR3A9DkIFzz7wQjII0jDaMEuUZH7QH2dOen6G3R91573cwBEhLcZOdS9y1wWx20f79k-LWbvKNpW1TUkjaFM7vFUr-voe62F2sAabJJcj7anlbuTTC9Kqhr_5Br_O-pObPcko0p8ww0Sh2pELJtRYYH6ahxN7FaI8x2ubX_ozRHlrRyz99PJT8yq0B_AjU-W7_UH73_o_sD48LqVw</addsrcrecordid><sourcetype>Open Access Repository</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>3107312924</pqid></control><display><type>article</type><title>Nepeta cataria L. (catnip) can serve as a chassis for the engineering of secondary metabolic pathways</title><source>MEDLINE</source><source>SpringerNature Journals</source><creator>Geissler, Marcus ; Neubauer, Christoph ; Sheludko, Yuriy V. ; Brückner, Adrian ; Warzecha, Heribert</creator><creatorcontrib>Geissler, Marcus ; Neubauer, Christoph ; Sheludko, Yuriy V. ; Brückner, Adrian ; Warzecha, Heribert</creatorcontrib><description>Objective
Evaluation of
Nepeta cataria
as a host with specific endogenous metabolite background for transient expression and metabolic engineering of secondary biosynthetic sequences.
Results
The reporter gene
gfp::licBM3
as well as three biosynthetic genes leading to the formation of the cannabinoid precursor olivetolic acid were adopted to the modular cloning standard GoldenBraid, transiently expressed in two chemotypes of
N. cataria
and compared to
Nicotiana benthamiana
. To estimate the expression efficiency in both hosts, quantification of the reporter activity was carried out with a sensitive and specific lichenase assay. While
N.
benthamiana
exhibited lichenase activity of 676 ± 94 μmol g
−1
s
−1
,
N. cataria
cultivar ‘1000’, and the cultivar ‘Citriodora’ showed an activity of 37 ± 8 μmol g
−1
s
−1
and 18 ± 4 μmol g
−1
s
−1
, respectively. Further, combinatorial expression of genes involved in cannabinoid biosynthetic pathway
acyl-activating enzyme 1
(
aae1
),
olivetol synthase
(
ols
) and
olivetolic acid cyclase
(
oac
) in
N. cataria
cv. resulted presumably in the in vivo production of olivetolic acid glycosides.
Conclusion
Nepeta cataria
is amenable to
Agrobacterium
-mediated transient expression and could serve as a novel chassis for the engineering of secondary metabolic pathways and transient evaluation of heterologous genes.</description><identifier>ISSN: 0141-5492</identifier><identifier>ISSN: 1573-6776</identifier><identifier>EISSN: 1573-6776</identifier><identifier>DOI: 10.1007/s10529-024-03489-w</identifier><identifier>PMID: 38717662</identifier><language>eng</language><publisher>Dordrecht: Springer Netherlands</publisher><subject>Applied Microbiology ; Biochemistry ; Biomedical and Life Sciences ; Biosynthetic Pathways - genetics ; Biotechnology ; Cannabinoids ; Cloning ; Combinatorial analysis ; Cultivars ; Gene sequencing ; Genes ; Genes, Reporter ; Glycosides ; Life Sciences ; Metabolic engineering ; Metabolic Engineering - methods ; Metabolic Networks and Pathways - genetics ; Metabolic pathways ; Metabolism ; Metabolites ; Microbiology ; Modular engineering ; Nepeta - chemistry ; Nepeta - genetics ; Nepeta - metabolism ; Nepeta cataria ; Nicotiana - genetics ; Nicotiana - metabolism ; Original Research Paper ; Plants, Genetically Modified - genetics ; Reporter gene</subject><ispartof>Biotechnology letters, 2024-10, Vol.46 (5), p.843-850</ispartof><rights>The Author(s) 2024</rights><rights>2024. The Author(s).</rights><rights>The Author(s) 2024. This work is published under http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>The Author(s) 2024 2024</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><cites>FETCH-LOGICAL-c426t-26d42fdbbe6040242ecdaa50943bddc5c33b2b4421e5921be6106b23d7be58183</cites><orcidid>0000-0001-8378-9243</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1007/s10529-024-03489-w$$EPDF$$P50$$Gspringer$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s10529-024-03489-w$$EHTML$$P50$$Gspringer$$Hfree_for_read</linktohtml><link.rule.ids>230,315,781,785,886,27929,27930,41493,42562,51324</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/38717662$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Geissler, Marcus</creatorcontrib><creatorcontrib>Neubauer, Christoph</creatorcontrib><creatorcontrib>Sheludko, Yuriy V.</creatorcontrib><creatorcontrib>Brückner, Adrian</creatorcontrib><creatorcontrib>Warzecha, Heribert</creatorcontrib><title>Nepeta cataria L. (catnip) can serve as a chassis for the engineering of secondary metabolic pathways</title><title>Biotechnology letters</title><addtitle>Biotechnol Lett</addtitle><addtitle>Biotechnol Lett</addtitle><description>Objective
Evaluation of
Nepeta cataria
as a host with specific endogenous metabolite background for transient expression and metabolic engineering of secondary biosynthetic sequences.
Results
The reporter gene
gfp::licBM3
as well as three biosynthetic genes leading to the formation of the cannabinoid precursor olivetolic acid were adopted to the modular cloning standard GoldenBraid, transiently expressed in two chemotypes of
N. cataria
and compared to
Nicotiana benthamiana
. To estimate the expression efficiency in both hosts, quantification of the reporter activity was carried out with a sensitive and specific lichenase assay. While
N.
benthamiana
exhibited lichenase activity of 676 ± 94 μmol g
−1
s
−1
,
N. cataria
cultivar ‘1000’, and the cultivar ‘Citriodora’ showed an activity of 37 ± 8 μmol g
−1
s
−1
and 18 ± 4 μmol g
−1
s
−1
, respectively. Further, combinatorial expression of genes involved in cannabinoid biosynthetic pathway
acyl-activating enzyme 1
(
aae1
),
olivetol synthase
(
ols
) and
olivetolic acid cyclase
(
oac
) in
N. cataria
cv. resulted presumably in the in vivo production of olivetolic acid glycosides.
Conclusion
Nepeta cataria
is amenable to
Agrobacterium
-mediated transient expression and could serve as a novel chassis for the engineering of secondary metabolic pathways and transient evaluation of heterologous genes.</description><subject>Applied Microbiology</subject><subject>Biochemistry</subject><subject>Biomedical and Life Sciences</subject><subject>Biosynthetic Pathways - genetics</subject><subject>Biotechnology</subject><subject>Cannabinoids</subject><subject>Cloning</subject><subject>Combinatorial analysis</subject><subject>Cultivars</subject><subject>Gene sequencing</subject><subject>Genes</subject><subject>Genes, Reporter</subject><subject>Glycosides</subject><subject>Life Sciences</subject><subject>Metabolic engineering</subject><subject>Metabolic Engineering - methods</subject><subject>Metabolic Networks and Pathways - genetics</subject><subject>Metabolic pathways</subject><subject>Metabolism</subject><subject>Metabolites</subject><subject>Microbiology</subject><subject>Modular engineering</subject><subject>Nepeta - chemistry</subject><subject>Nepeta - genetics</subject><subject>Nepeta - metabolism</subject><subject>Nepeta cataria</subject><subject>Nicotiana - genetics</subject><subject>Nicotiana - metabolism</subject><subject>Original Research Paper</subject><subject>Plants, Genetically Modified - genetics</subject><subject>Reporter gene</subject><issn>0141-5492</issn><issn>1573-6776</issn><issn>1573-6776</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2024</creationdate><recordtype>article</recordtype><sourceid>C6C</sourceid><sourceid>EIF</sourceid><recordid>eNp9kctuFDEQRS0EIpPAD7BAltiERQe_3V4hFEFAGsEG1pbtrp5x1GM3dk9G-XscJoTHgpUt-9StunURekHJBSVEv6mUSGY6wkRHuOhNd3iEVlRq3imt1WO0IlTQTgrDTtBprdeEEKOJfopOeK-pVoqtEHyGGRaHg1tciQ6vL_B5u6c4v25vCVcoN4BdxQ3ZulpjxWMueNkChrSJCaDEtMF5bGTIaXDlFu-aoM9TDHh2y_bgbusz9GR0U4Xn9-cZ-vbh_dfLj936y9Wny3frLgimlo6pQbBx8B4UEc0WgzA4J4kR3A9DkIFzz7wQjII0jDaMEuUZH7QH2dOen6G3R91573cwBEhLcZOdS9y1wWx20f79k-LWbvKNpW1TUkjaFM7vFUr-voe62F2sAabJJcj7anlbuTTC9Kqhr_5Br_O-pObPcko0p8ww0Sh2pELJtRYYH6ahxN7FaI8x2ubX_ozRHlrRyz99PJT8yq0B_AjU-W7_UH73_o_sD48LqVw</recordid><startdate>20241001</startdate><enddate>20241001</enddate><creator>Geissler, Marcus</creator><creator>Neubauer, Christoph</creator><creator>Sheludko, Yuriy V.</creator><creator>Brückner, Adrian</creator><creator>Warzecha, Heribert</creator><general>Springer Netherlands</general><general>Springer Nature B.V</general><scope>C6C</scope><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>7QL</scope><scope>7QR</scope><scope>7T7</scope><scope>7TB</scope><scope>7U5</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>K9.</scope><scope>L7M</scope><scope>M7N</scope><scope>P64</scope><scope>7X8</scope><scope>5PM</scope><orcidid>https://orcid.org/0000-0001-8378-9243</orcidid></search><sort><creationdate>20241001</creationdate><title>Nepeta cataria L. (catnip) can serve as a chassis for the engineering of secondary metabolic pathways</title><author>Geissler, Marcus ; Neubauer, Christoph ; Sheludko, Yuriy V. ; Brückner, Adrian ; Warzecha, Heribert</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c426t-26d42fdbbe6040242ecdaa50943bddc5c33b2b4421e5921be6106b23d7be58183</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2024</creationdate><topic>Applied Microbiology</topic><topic>Biochemistry</topic><topic>Biomedical and Life Sciences</topic><topic>Biosynthetic Pathways - genetics</topic><topic>Biotechnology</topic><topic>Cannabinoids</topic><topic>Cloning</topic><topic>Combinatorial analysis</topic><topic>Cultivars</topic><topic>Gene sequencing</topic><topic>Genes</topic><topic>Genes, Reporter</topic><topic>Glycosides</topic><topic>Life Sciences</topic><topic>Metabolic engineering</topic><topic>Metabolic Engineering - methods</topic><topic>Metabolic Networks and Pathways - genetics</topic><topic>Metabolic pathways</topic><topic>Metabolism</topic><topic>Metabolites</topic><topic>Microbiology</topic><topic>Modular engineering</topic><topic>Nepeta - chemistry</topic><topic>Nepeta - genetics</topic><topic>Nepeta - metabolism</topic><topic>Nepeta cataria</topic><topic>Nicotiana - genetics</topic><topic>Nicotiana - metabolism</topic><topic>Original Research Paper</topic><topic>Plants, Genetically Modified - genetics</topic><topic>Reporter gene</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Geissler, Marcus</creatorcontrib><creatorcontrib>Neubauer, Christoph</creatorcontrib><creatorcontrib>Sheludko, Yuriy V.</creatorcontrib><creatorcontrib>Brückner, Adrian</creatorcontrib><creatorcontrib>Warzecha, Heribert</creatorcontrib><collection>Springer Nature OA/Free Journals</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Chemoreception Abstracts</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Mechanical & Transportation Engineering Abstracts</collection><collection>Solid State and Superconductivity Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Advanced Technologies Database with Aerospace</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Biotechnology letters</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Geissler, Marcus</au><au>Neubauer, Christoph</au><au>Sheludko, Yuriy V.</au><au>Brückner, Adrian</au><au>Warzecha, Heribert</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Nepeta cataria L. (catnip) can serve as a chassis for the engineering of secondary metabolic pathways</atitle><jtitle>Biotechnology letters</jtitle><stitle>Biotechnol Lett</stitle><addtitle>Biotechnol Lett</addtitle><date>2024-10-01</date><risdate>2024</risdate><volume>46</volume><issue>5</issue><spage>843</spage><epage>850</epage><pages>843-850</pages><issn>0141-5492</issn><issn>1573-6776</issn><eissn>1573-6776</eissn><abstract>Objective
Evaluation of
Nepeta cataria
as a host with specific endogenous metabolite background for transient expression and metabolic engineering of secondary biosynthetic sequences.
Results
The reporter gene
gfp::licBM3
as well as three biosynthetic genes leading to the formation of the cannabinoid precursor olivetolic acid were adopted to the modular cloning standard GoldenBraid, transiently expressed in two chemotypes of
N. cataria
and compared to
Nicotiana benthamiana
. To estimate the expression efficiency in both hosts, quantification of the reporter activity was carried out with a sensitive and specific lichenase assay. While
N.
benthamiana
exhibited lichenase activity of 676 ± 94 μmol g
−1
s
−1
,
N. cataria
cultivar ‘1000’, and the cultivar ‘Citriodora’ showed an activity of 37 ± 8 μmol g
−1
s
−1
and 18 ± 4 μmol g
−1
s
−1
, respectively. Further, combinatorial expression of genes involved in cannabinoid biosynthetic pathway
acyl-activating enzyme 1
(
aae1
),
olivetol synthase
(
ols
) and
olivetolic acid cyclase
(
oac
) in
N. cataria
cv. resulted presumably in the in vivo production of olivetolic acid glycosides.
Conclusion
Nepeta cataria
is amenable to
Agrobacterium
-mediated transient expression and could serve as a novel chassis for the engineering of secondary metabolic pathways and transient evaluation of heterologous genes.</abstract><cop>Dordrecht</cop><pub>Springer Netherlands</pub><pmid>38717662</pmid><doi>10.1007/s10529-024-03489-w</doi><tpages>8</tpages><orcidid>https://orcid.org/0000-0001-8378-9243</orcidid><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0141-5492 |
| ispartof | Biotechnology letters, 2024-10, Vol.46 (5), p.843-850 |
| issn | 0141-5492 1573-6776 1573-6776 |
| language | eng |
| recordid | cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_11415451 |
| source | MEDLINE; SpringerNature Journals |
| subjects | Applied Microbiology Biochemistry Biomedical and Life Sciences Biosynthetic Pathways - genetics Biotechnology Cannabinoids Cloning Combinatorial analysis Cultivars Gene sequencing Genes Genes, Reporter Glycosides Life Sciences Metabolic engineering Metabolic Engineering - methods Metabolic Networks and Pathways - genetics Metabolic pathways Metabolism Metabolites Microbiology Modular engineering Nepeta - chemistry Nepeta - genetics Nepeta - metabolism Nepeta cataria Nicotiana - genetics Nicotiana - metabolism Original Research Paper Plants, Genetically Modified - genetics Reporter gene |
| title | Nepeta cataria L. (catnip) can serve as a chassis for the engineering of secondary metabolic pathways |
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