Electrochemical Method for the Detection of Lipase Activity
A novel electrochemical technique for the general assay of lipase activity is described. The method utilizes a solid-supported lipase substrate, which is formed by dripping and drying a small amount of an ethanol solution of 9-(5‘-ferrocenylpentanoyloxy)nonyl disulfide (FPONDS) onto gold modified by...
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| Veröffentlicht in: | Analytical chemistry (Washington) 2005-04, Vol.77 (8), p.2632-2636 |
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| container_title | Analytical chemistry (Washington) |
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| creator | Valincius, Gintaras Ignatjev, Ilja Niaura, Gediminas Kažemėkaitė, Marytė Talaikytė, Zita Razumas, Valdemaras Svendsen, Allan |
| description | A novel electrochemical technique for the general assay of lipase activity is described. The method utilizes a solid-supported lipase substrate, which is formed by dripping and drying a small amount of an ethanol solution of 9-(5‘-ferrocenylpentanoyloxy)nonyl disulfide (FPONDS) onto gold modified by a hexanethiol self-assembled monolayer. The redox ferrocene group of FPONDS generates the electrochemical signal, the intensity of which is proportional to the number of FPONDS molecules at the interface. Electrochemical and surface-enhanced infrared absorption spectroscopic data, as well as control experiments with an engineered, deactivated mutant enzyme, demonstrate that the wild-type lipase from Thermomyces lanuginosus is capable of cleaving the ester bonds of FPONDS molecules via an enzymatic hydrolysis mechanism, which includes the adsorption of the lipase onto the substrate surface. The hydrolysis liberates the ferrocene groups from the interface triggering a decay of the electrochemical redox signal. The rate of the electrochemical signal decrease is proportional to the lipase activity/concentration. These data suggest a general method for the direct measure of enzymatic activity of lipases. |
| doi_str_mv | 10.1021/ac048230+ |
| format | Article |
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The method utilizes a solid-supported lipase substrate, which is formed by dripping and drying a small amount of an ethanol solution of 9-(5‘-ferrocenylpentanoyloxy)nonyl disulfide (FPONDS) onto gold modified by a hexanethiol self-assembled monolayer. The redox ferrocene group of FPONDS generates the electrochemical signal, the intensity of which is proportional to the number of FPONDS molecules at the interface. Electrochemical and surface-enhanced infrared absorption spectroscopic data, as well as control experiments with an engineered, deactivated mutant enzyme, demonstrate that the wild-type lipase from Thermomyces lanuginosus is capable of cleaving the ester bonds of FPONDS molecules via an enzymatic hydrolysis mechanism, which includes the adsorption of the lipase onto the substrate surface. The hydrolysis liberates the ferrocene groups from the interface triggering a decay of the electrochemical redox signal. The rate of the electrochemical signal decrease is proportional to the lipase activity/concentration. These data suggest a general method for the direct measure of enzymatic activity of lipases.</description><identifier>ISSN: 0003-2700</identifier><identifier>EISSN: 1520-6882</identifier><identifier>DOI: 10.1021/ac048230+</identifier><identifier>PMID: 15828803</identifier><identifier>CODEN: ANCHAM</identifier><language>eng</language><publisher>Washington, DC: American Chemical Society</publisher><subject>Analytical chemistry ; Chemical bonds ; Chemistry ; Disulfides - chemistry ; Disulfides - metabolism ; Electrochemical methods ; Electrochemistry - methods ; Enzymes ; Exact sciences and technology ; Ferrous Compounds - chemistry ; Ferrous Compounds - metabolism ; Kinetics ; Lipase - analysis ; Lipase - genetics ; Lipase - metabolism ; Metallocenes ; Mitosporic Fungi - enzymology ; Mutation ; Oxidation-Reduction</subject><ispartof>Analytical chemistry (Washington), 2005-04, Vol.77 (8), p.2632-2636</ispartof><rights>2005 INIST-CNRS</rights><rights>Copyright American Chemical Society Apr 15, 2005</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-a405t-63eeb9252e5b592f9c925f8dd19ea6775add92ad258a126beea4a9769b8d47d3</citedby><cites>FETCH-LOGICAL-a405t-63eeb9252e5b592f9c925f8dd19ea6775add92ad258a126beea4a9769b8d47d3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://pubs.acs.org/doi/pdf/10.1021/ac048230+$$EPDF$$P50$$Gacs$$H</linktopdf><linktohtml>$$Uhttps://pubs.acs.org/doi/10.1021/ac048230+$$EHTML$$P50$$Gacs$$H</linktohtml><link.rule.ids>314,780,784,2765,27076,27924,27925,56738,56788</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=16708045$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/15828803$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Valincius, Gintaras</creatorcontrib><creatorcontrib>Ignatjev, Ilja</creatorcontrib><creatorcontrib>Niaura, Gediminas</creatorcontrib><creatorcontrib>Kažemėkaitė, Marytė</creatorcontrib><creatorcontrib>Talaikytė, Zita</creatorcontrib><creatorcontrib>Razumas, Valdemaras</creatorcontrib><creatorcontrib>Svendsen, Allan</creatorcontrib><title>Electrochemical Method for the Detection of Lipase Activity</title><title>Analytical chemistry (Washington)</title><addtitle>Anal. Chem</addtitle><description>A novel electrochemical technique for the general assay of lipase activity is described. The method utilizes a solid-supported lipase substrate, which is formed by dripping and drying a small amount of an ethanol solution of 9-(5‘-ferrocenylpentanoyloxy)nonyl disulfide (FPONDS) onto gold modified by a hexanethiol self-assembled monolayer. The redox ferrocene group of FPONDS generates the electrochemical signal, the intensity of which is proportional to the number of FPONDS molecules at the interface. Electrochemical and surface-enhanced infrared absorption spectroscopic data, as well as control experiments with an engineered, deactivated mutant enzyme, demonstrate that the wild-type lipase from Thermomyces lanuginosus is capable of cleaving the ester bonds of FPONDS molecules via an enzymatic hydrolysis mechanism, which includes the adsorption of the lipase onto the substrate surface. The hydrolysis liberates the ferrocene groups from the interface triggering a decay of the electrochemical redox signal. The rate of the electrochemical signal decrease is proportional to the lipase activity/concentration. These data suggest a general method for the direct measure of enzymatic activity of lipases.</description><subject>Analytical chemistry</subject><subject>Chemical bonds</subject><subject>Chemistry</subject><subject>Disulfides - chemistry</subject><subject>Disulfides - metabolism</subject><subject>Electrochemical methods</subject><subject>Electrochemistry - methods</subject><subject>Enzymes</subject><subject>Exact sciences and technology</subject><subject>Ferrous Compounds - chemistry</subject><subject>Ferrous Compounds - metabolism</subject><subject>Kinetics</subject><subject>Lipase - analysis</subject><subject>Lipase - genetics</subject><subject>Lipase - metabolism</subject><subject>Metallocenes</subject><subject>Mitosporic Fungi - enzymology</subject><subject>Mutation</subject><subject>Oxidation-Reduction</subject><issn>0003-2700</issn><issn>1520-6882</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2005</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpl0F1rFDEUBuAglXatvfAPyFCqCGXsSTLJZPCqrPUDVpR2kd6FM8kZdursZk1mxf57U3Z1wV6F5Dy8nLyMveDwloPgF-igMkLC-RM24UpAqY0RB2wCALIUNcARe5bSHQDnwPUhO-LKCGNATti7q4HcGINb0LJ3OBRfaFwEX3QhFuOCivc05nkfVkXoilm_xkTFZX741Y_3z9nTDodEJ7vzmM0_XM2nn8rZ14-fp5ezEitQY6klUdsIJUi1qhFd4_KlM97zhlDXtULvG4FeKINc6JYIK2xq3bTGV7WXx-z1NnYdw88NpdEu--RoGHBFYZNsjqhkJXWGp__Bu7CJq7yaFbw2RmsuM3qzRS6GlCJ1dh37JcZ7y8E-tGn_tpnpy13epl2S38Ndexm82gFMubsu4sr1ae90DQYqlV25dX0a6fe_OcYfeXlZKzv_dmOvb25nt830u33IPdt6dGn_h0f7_QGIu5TQ</recordid><startdate>20050415</startdate><enddate>20050415</enddate><creator>Valincius, Gintaras</creator><creator>Ignatjev, Ilja</creator><creator>Niaura, Gediminas</creator><creator>Kažemėkaitė, Marytė</creator><creator>Talaikytė, Zita</creator><creator>Razumas, Valdemaras</creator><creator>Svendsen, Allan</creator><general>American Chemical Society</general><scope>BSCLL</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>7QF</scope><scope>7QO</scope><scope>7QQ</scope><scope>7SC</scope><scope>7SE</scope><scope>7SP</scope><scope>7SR</scope><scope>7TA</scope><scope>7TB</scope><scope>7TM</scope><scope>7U5</scope><scope>7U7</scope><scope>7U9</scope><scope>8BQ</scope><scope>8FD</scope><scope>C1K</scope><scope>F28</scope><scope>FR3</scope><scope>H8D</scope><scope>H8G</scope><scope>H94</scope><scope>JG9</scope><scope>JQ2</scope><scope>KR7</scope><scope>L7M</scope><scope>L~C</scope><scope>L~D</scope><scope>P64</scope><scope>7X8</scope></search><sort><creationdate>20050415</creationdate><title>Electrochemical Method for the Detection of Lipase Activity</title><author>Valincius, Gintaras ; Ignatjev, Ilja ; Niaura, Gediminas ; Kažemėkaitė, Marytė ; Talaikytė, Zita ; Razumas, Valdemaras ; Svendsen, Allan</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-a405t-63eeb9252e5b592f9c925f8dd19ea6775add92ad258a126beea4a9769b8d47d3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2005</creationdate><topic>Analytical chemistry</topic><topic>Chemical bonds</topic><topic>Chemistry</topic><topic>Disulfides - chemistry</topic><topic>Disulfides - metabolism</topic><topic>Electrochemical methods</topic><topic>Electrochemistry - methods</topic><topic>Enzymes</topic><topic>Exact sciences and technology</topic><topic>Ferrous Compounds - chemistry</topic><topic>Ferrous Compounds - metabolism</topic><topic>Kinetics</topic><topic>Lipase - analysis</topic><topic>Lipase - genetics</topic><topic>Lipase - metabolism</topic><topic>Metallocenes</topic><topic>Mitosporic Fungi - enzymology</topic><topic>Mutation</topic><topic>Oxidation-Reduction</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Valincius, Gintaras</creatorcontrib><creatorcontrib>Ignatjev, Ilja</creatorcontrib><creatorcontrib>Niaura, Gediminas</creatorcontrib><creatorcontrib>Kažemėkaitė, Marytė</creatorcontrib><creatorcontrib>Talaikytė, Zita</creatorcontrib><creatorcontrib>Razumas, Valdemaras</creatorcontrib><creatorcontrib>Svendsen, Allan</creatorcontrib><collection>Istex</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>Aluminium Industry Abstracts</collection><collection>Biotechnology Research Abstracts</collection><collection>Ceramic Abstracts</collection><collection>Computer and Information Systems Abstracts</collection><collection>Corrosion Abstracts</collection><collection>Electronics & Communications Abstracts</collection><collection>Engineered Materials Abstracts</collection><collection>Materials Business File</collection><collection>Mechanical & Transportation Engineering Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Solid State and Superconductivity Abstracts</collection><collection>Toxicology Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>METADEX</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ANTE: Abstracts in New Technology & Engineering</collection><collection>Engineering Research Database</collection><collection>Aerospace Database</collection><collection>Copper Technical Reference Library</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Materials Research Database</collection><collection>ProQuest Computer Science Collection</collection><collection>Civil Engineering Abstracts</collection><collection>Advanced Technologies Database with Aerospace</collection><collection>Computer and Information Systems Abstracts Academic</collection><collection>Computer and Information Systems Abstracts Professional</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Analytical chemistry (Washington)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Valincius, Gintaras</au><au>Ignatjev, Ilja</au><au>Niaura, Gediminas</au><au>Kažemėkaitė, Marytė</au><au>Talaikytė, Zita</au><au>Razumas, Valdemaras</au><au>Svendsen, Allan</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Electrochemical Method for the Detection of Lipase Activity</atitle><jtitle>Analytical chemistry (Washington)</jtitle><addtitle>Anal. Chem</addtitle><date>2005-04-15</date><risdate>2005</risdate><volume>77</volume><issue>8</issue><spage>2632</spage><epage>2636</epage><pages>2632-2636</pages><issn>0003-2700</issn><eissn>1520-6882</eissn><coden>ANCHAM</coden><abstract>A novel electrochemical technique for the general assay of lipase activity is described. The method utilizes a solid-supported lipase substrate, which is formed by dripping and drying a small amount of an ethanol solution of 9-(5‘-ferrocenylpentanoyloxy)nonyl disulfide (FPONDS) onto gold modified by a hexanethiol self-assembled monolayer. The redox ferrocene group of FPONDS generates the electrochemical signal, the intensity of which is proportional to the number of FPONDS molecules at the interface. Electrochemical and surface-enhanced infrared absorption spectroscopic data, as well as control experiments with an engineered, deactivated mutant enzyme, demonstrate that the wild-type lipase from Thermomyces lanuginosus is capable of cleaving the ester bonds of FPONDS molecules via an enzymatic hydrolysis mechanism, which includes the adsorption of the lipase onto the substrate surface. The hydrolysis liberates the ferrocene groups from the interface triggering a decay of the electrochemical redox signal. The rate of the electrochemical signal decrease is proportional to the lipase activity/concentration. These data suggest a general method for the direct measure of enzymatic activity of lipases.</abstract><cop>Washington, DC</cop><pub>American Chemical Society</pub><pmid>15828803</pmid><doi>10.1021/ac048230+</doi><tpages>5</tpages></addata></record> |
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| subjects | Analytical chemistry Chemical bonds Chemistry Disulfides - chemistry Disulfides - metabolism Electrochemical methods Electrochemistry - methods Enzymes Exact sciences and technology Ferrous Compounds - chemistry Ferrous Compounds - metabolism Kinetics Lipase - analysis Lipase - genetics Lipase - metabolism Metallocenes Mitosporic Fungi - enzymology Mutation Oxidation-Reduction |
| title | Electrochemical Method for the Detection of Lipase Activity |
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