Quantitative analysis of isoprenoid diphosphate intermediates in recombinant and wild-type Escherichia coli strains
In biotechnology, the heterologous biosynthesis of isoprenoid compounds in Escherichia coli is a field of great interest and growth. In order to achieve higher isoprenoid yields in heterologous E. coli strains, it is necessary to quantify the pathway intermediates and adjust gene expression. In this...
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| Veröffentlicht in: | Applied microbiology and biotechnology 2008-11, Vol.81 (1), p.175-182 |
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| creator | Vallon, T Ghanegaonkar, S Vielhauer, O Müller, A Albermann, C Sprenger, G Reuss, M Lemuth, K |
| description | In biotechnology, the heterologous biosynthesis of isoprenoid compounds in Escherichia coli is a field of great interest and growth. In order to achieve higher isoprenoid yields in heterologous E. coli strains, it is necessary to quantify the pathway intermediates and adjust gene expression. In this study, we developed a precise and sensitive nonradioactive method for the simultaneous quantification of the isoprenoid precursors farnesyl diphosphate (FPP) and geranylgeranyl diphosphate (GGPP) in recombinant and wild-type E. coli cells. The method is based on the dephosphorylation of FPP and GGPP into the respective alcohols and involves their in situ extraction followed by separation and detection using gas chromatography-mass spectrometry. The integration of a geranylgeranyl diphosphate synthase gene into the E. coli chromosome leads to the accumulation of GGPP, generating quantities as high as those achieved with a multicopy expression vector. |
| doi_str_mv | 10.1007/s00253-008-1707-8 |
| format | Article |
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In order to achieve higher isoprenoid yields in heterologous E. coli strains, it is necessary to quantify the pathway intermediates and adjust gene expression. In this study, we developed a precise and sensitive nonradioactive method for the simultaneous quantification of the isoprenoid precursors farnesyl diphosphate (FPP) and geranylgeranyl diphosphate (GGPP) in recombinant and wild-type E. coli cells. The method is based on the dephosphorylation of FPP and GGPP into the respective alcohols and involves their in situ extraction followed by separation and detection using gas chromatography-mass spectrometry. The integration of a geranylgeranyl diphosphate synthase gene into the E. coli chromosome leads to the accumulation of GGPP, generating quantities as high as those achieved with a multicopy expression vector.</description><identifier>ISSN: 0175-7598</identifier><identifier>EISSN: 1432-0614</identifier><identifier>DOI: 10.1007/s00253-008-1707-8</identifier><identifier>PMID: 18813922</identifier><identifier>CODEN: AMBIDG</identifier><language>eng</language><publisher>Berlin/Heidelberg: Berlin/Heidelberg : Springer-Verlag</publisher><subject>Alcohols - metabolism ; Biological and medical sciences ; Biomass ; Bioreactors - microbiology ; Biosynthesis ; Biotechnology ; Carotenoids ; Chromosomal integration ; Chromosomes ; Cloning ; E coli ; Enzymes ; Escherichia coli ; Escherichia coli - chemistry ; Escherichia coli - genetics ; Escherichia coli - metabolism ; Fermentation ; Fundamental and applied biological sciences. Psychology ; Gas chromatography ; Gene Expression ; Genetic Engineering ; Genetic recombination ; Geranylgeranyl diphosphate synthase ; Heterologous expression ; Isoprenoid diphosphate ; Life Sciences ; Mass spectrometry ; Methods ; Microbial Genetics and Genomics ; Microbiology ; Plasmids ; Polyisoprenyl Phosphates - analysis ; Polyisoprenyl Phosphates - isolation & purification ; Polyisoprenyl Phosphates - metabolism ; Quantitative analysis ; Studies</subject><ispartof>Applied microbiology and biotechnology, 2008-11, Vol.81 (1), p.175-182</ispartof><rights>Springer-Verlag 2008</rights><rights>2009 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c520t-33e051e48bfe050342e3cec8d18d6534f5e5fadf5900ac72da41bc80ea67c6613</citedby><cites>FETCH-LOGICAL-c520t-33e051e48bfe050342e3cec8d18d6534f5e5fadf5900ac72da41bc80ea67c6613</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1007/s00253-008-1707-8$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s00253-008-1707-8$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,780,784,27924,27925,41488,42557,51319</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=21017819$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/18813922$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Vallon, T</creatorcontrib><creatorcontrib>Ghanegaonkar, S</creatorcontrib><creatorcontrib>Vielhauer, O</creatorcontrib><creatorcontrib>Müller, A</creatorcontrib><creatorcontrib>Albermann, C</creatorcontrib><creatorcontrib>Sprenger, G</creatorcontrib><creatorcontrib>Reuss, M</creatorcontrib><creatorcontrib>Lemuth, K</creatorcontrib><title>Quantitative analysis of isoprenoid diphosphate intermediates in recombinant and wild-type Escherichia coli strains</title><title>Applied microbiology and biotechnology</title><addtitle>Appl Microbiol Biotechnol</addtitle><addtitle>Appl Microbiol Biotechnol</addtitle><description>In biotechnology, the heterologous biosynthesis of isoprenoid compounds in Escherichia coli is a field of great interest and growth. In order to achieve higher isoprenoid yields in heterologous E. coli strains, it is necessary to quantify the pathway intermediates and adjust gene expression. In this study, we developed a precise and sensitive nonradioactive method for the simultaneous quantification of the isoprenoid precursors farnesyl diphosphate (FPP) and geranylgeranyl diphosphate (GGPP) in recombinant and wild-type E. coli cells. The method is based on the dephosphorylation of FPP and GGPP into the respective alcohols and involves their in situ extraction followed by separation and detection using gas chromatography-mass spectrometry. The integration of a geranylgeranyl diphosphate synthase gene into the E. coli chromosome leads to the accumulation of GGPP, generating quantities as high as those achieved with a multicopy expression vector.</description><subject>Alcohols - metabolism</subject><subject>Biological and medical sciences</subject><subject>Biomass</subject><subject>Bioreactors - microbiology</subject><subject>Biosynthesis</subject><subject>Biotechnology</subject><subject>Carotenoids</subject><subject>Chromosomal integration</subject><subject>Chromosomes</subject><subject>Cloning</subject><subject>E coli</subject><subject>Enzymes</subject><subject>Escherichia coli</subject><subject>Escherichia coli - chemistry</subject><subject>Escherichia coli - genetics</subject><subject>Escherichia coli - metabolism</subject><subject>Fermentation</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gas chromatography</subject><subject>Gene Expression</subject><subject>Genetic Engineering</subject><subject>Genetic recombination</subject><subject>Geranylgeranyl diphosphate synthase</subject><subject>Heterologous expression</subject><subject>Isoprenoid diphosphate</subject><subject>Life Sciences</subject><subject>Mass spectrometry</subject><subject>Methods</subject><subject>Microbial Genetics and Genomics</subject><subject>Microbiology</subject><subject>Plasmids</subject><subject>Polyisoprenyl Phosphates - analysis</subject><subject>Polyisoprenyl Phosphates - isolation & purification</subject><subject>Polyisoprenyl Phosphates - metabolism</subject><subject>Quantitative analysis</subject><subject>Studies</subject><issn>0175-7598</issn><issn>1432-0614</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2008</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNqFkc1u1DAURi0EokPhAdhAhFR2gXvt-CfLqmoBqRJC0LXlcZyOq0wcfBPQvD0uGVGJBaxsy-f7bN3D2EuEdwig3xMAl6IGMDVq0LV5xDbYCF6DwuYx2wBqWWvZmhP2jOgOALlR6ik7QWNQtJxvGH1Z3DjH2c3xR6jc6IYDRapSX0VKUw5jil3VxWmXaNq5OVRxnEPehy6WA5VTlYNP-20cS03Jd9XPOHT1fJhCdUl-F3L0u-gqn4ZY0ZxdHOk5e9K7gcKL43rKbq4uv118rK8_f_h0cX5de8lhroUIIDE0ZtuXDYiGB-GDNx2aTknR9DLI3nW9bAGc17xzDW69geCU9kqhOGVv194pp-9LoNnuI_kwDG4MaSGrWtWKFtV_QY5cNkLpAr75C7xLSy5DKwxvFSr1G8IV8jkR5dDbKce9yweLYO-92dWbLd7svTdrSubVsXjZluE-JI6iCnB2BBx5N_TZjT7SH45jcW2wLRxfOSpX423IDz_81-uv11DvknW3uRTffOWAAlBq3XIUvwAT7LrA</recordid><startdate>20081101</startdate><enddate>20081101</enddate><creator>Vallon, T</creator><creator>Ghanegaonkar, S</creator><creator>Vielhauer, O</creator><creator>Müller, A</creator><creator>Albermann, C</creator><creator>Sprenger, G</creator><creator>Reuss, M</creator><creator>Lemuth, K</creator><general>Berlin/Heidelberg : Springer-Verlag</general><general>Springer Berlin Heidelberg</general><general>Springer</general><general>Springer Nature B.V</general><scope>FBQ</scope><scope>IQODW</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>3V.</scope><scope>7QL</scope><scope>7T7</scope><scope>7WY</scope><scope>7WZ</scope><scope>7X7</scope><scope>7XB</scope><scope>87Z</scope><scope>88A</scope><scope>88E</scope><scope>88I</scope><scope>8AO</scope><scope>8FD</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>8FL</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BEZIV</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FRNLG</scope><scope>FYUFA</scope><scope>F~G</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>HCIFZ</scope><scope>K60</scope><scope>K6~</scope><scope>K9.</scope><scope>L.-</scope><scope>LK8</scope><scope>M0C</scope><scope>M0S</scope><scope>M1P</scope><scope>M2P</scope><scope>M7N</scope><scope>M7P</scope><scope>P64</scope><scope>PQBIZ</scope><scope>PQBZA</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>Q9U</scope><scope>7QO</scope><scope>7X8</scope></search><sort><creationdate>20081101</creationdate><title>Quantitative analysis of isoprenoid diphosphate intermediates in recombinant and wild-type Escherichia coli strains</title><author>Vallon, T ; Ghanegaonkar, S ; Vielhauer, O ; Müller, A ; Albermann, C ; Sprenger, G ; Reuss, M ; Lemuth, K</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c520t-33e051e48bfe050342e3cec8d18d6534f5e5fadf5900ac72da41bc80ea67c6613</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2008</creationdate><topic>Alcohols - metabolism</topic><topic>Biological and medical sciences</topic><topic>Biomass</topic><topic>Bioreactors - microbiology</topic><topic>Biosynthesis</topic><topic>Biotechnology</topic><topic>Carotenoids</topic><topic>Chromosomal integration</topic><topic>Chromosomes</topic><topic>Cloning</topic><topic>E coli</topic><topic>Enzymes</topic><topic>Escherichia coli</topic><topic>Escherichia coli - chemistry</topic><topic>Escherichia coli - genetics</topic><topic>Escherichia coli - metabolism</topic><topic>Fermentation</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Gas chromatography</topic><topic>Gene Expression</topic><topic>Genetic Engineering</topic><topic>Genetic recombination</topic><topic>Geranylgeranyl diphosphate synthase</topic><topic>Heterologous expression</topic><topic>Isoprenoid diphosphate</topic><topic>Life Sciences</topic><topic>Mass spectrometry</topic><topic>Methods</topic><topic>Microbial Genetics and Genomics</topic><topic>Microbiology</topic><topic>Plasmids</topic><topic>Polyisoprenyl Phosphates - analysis</topic><topic>Polyisoprenyl Phosphates - isolation & purification</topic><topic>Polyisoprenyl Phosphates - metabolism</topic><topic>Quantitative analysis</topic><topic>Studies</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Vallon, T</creatorcontrib><creatorcontrib>Ghanegaonkar, S</creatorcontrib><creatorcontrib>Vielhauer, O</creatorcontrib><creatorcontrib>Müller, A</creatorcontrib><creatorcontrib>Albermann, C</creatorcontrib><creatorcontrib>Sprenger, G</creatorcontrib><creatorcontrib>Reuss, M</creatorcontrib><creatorcontrib>Lemuth, K</creatorcontrib><collection>AGRIS</collection><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Access via ABI/INFORM (ProQuest)</collection><collection>ABI/INFORM Global (PDF only)</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>ABI/INFORM Global (Alumni Edition)</collection><collection>Biology Database (Alumni Edition)</collection><collection>Medical Database (Alumni Edition)</collection><collection>Science Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech 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>ABI/INFORM Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest Central UK/Ireland</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Business Premium Collection</collection><collection>Natural Science Collection</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central Korea</collection><collection>Engineering Research Database</collection><collection>Business Premium Collection (Alumni)</collection><collection>Health Research Premium Collection</collection><collection>ABI/INFORM Global (Corporate)</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Business Collection (Alumni Edition)</collection><collection>ProQuest Business Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>ABI/INFORM Professional Advanced</collection><collection>ProQuest Biological Science Collection</collection><collection>ABI/INFORM Global</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Science Database</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biological Science Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>One Business (ProQuest)</collection><collection>ProQuest One Business (Alumni)</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 Basic</collection><collection>Biotechnology Research Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Applied microbiology and biotechnology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Vallon, T</au><au>Ghanegaonkar, S</au><au>Vielhauer, O</au><au>Müller, A</au><au>Albermann, C</au><au>Sprenger, G</au><au>Reuss, M</au><au>Lemuth, K</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Quantitative analysis of isoprenoid diphosphate intermediates in recombinant and wild-type Escherichia coli strains</atitle><jtitle>Applied microbiology and biotechnology</jtitle><stitle>Appl Microbiol Biotechnol</stitle><addtitle>Appl Microbiol Biotechnol</addtitle><date>2008-11-01</date><risdate>2008</risdate><volume>81</volume><issue>1</issue><spage>175</spage><epage>182</epage><pages>175-182</pages><issn>0175-7598</issn><eissn>1432-0614</eissn><coden>AMBIDG</coden><abstract>In biotechnology, the heterologous biosynthesis of isoprenoid compounds in Escherichia coli is a field of great interest and growth. In order to achieve higher isoprenoid yields in heterologous E. coli strains, it is necessary to quantify the pathway intermediates and adjust gene expression. In this study, we developed a precise and sensitive nonradioactive method for the simultaneous quantification of the isoprenoid precursors farnesyl diphosphate (FPP) and geranylgeranyl diphosphate (GGPP) in recombinant and wild-type E. coli cells. The method is based on the dephosphorylation of FPP and GGPP into the respective alcohols and involves their in situ extraction followed by separation and detection using gas chromatography-mass spectrometry. The integration of a geranylgeranyl diphosphate synthase gene into the E. coli chromosome leads to the accumulation of GGPP, generating quantities as high as those achieved with a multicopy expression vector.</abstract><cop>Berlin/Heidelberg</cop><pub>Berlin/Heidelberg : Springer-Verlag</pub><pmid>18813922</pmid><doi>10.1007/s00253-008-1707-8</doi><tpages>8</tpages></addata></record> |
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| subjects | Alcohols - metabolism Biological and medical sciences Biomass Bioreactors - microbiology Biosynthesis Biotechnology Carotenoids Chromosomal integration Chromosomes Cloning E coli Enzymes Escherichia coli Escherichia coli - chemistry Escherichia coli - genetics Escherichia coli - metabolism Fermentation Fundamental and applied biological sciences. Psychology Gas chromatography Gene Expression Genetic Engineering Genetic recombination Geranylgeranyl diphosphate synthase Heterologous expression Isoprenoid diphosphate Life Sciences Mass spectrometry Methods Microbial Genetics and Genomics Microbiology Plasmids Polyisoprenyl Phosphates - analysis Polyisoprenyl Phosphates - isolation & purification Polyisoprenyl Phosphates - metabolism Quantitative analysis Studies |
| title | Quantitative analysis of isoprenoid diphosphate intermediates in recombinant and wild-type Escherichia coli strains |
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