Affinity purification of bacterial luciferase and NAD(P)H:FMN oxidoreductases by FMN-sepharose for analytical applications
A modified purification method for bacterial luciferases and NAD(P)H:FMN oxidoreductases is described which uses FMN-Sepharose alone or coupled to DEAE ion exchange chromatography for the simultaneous purification of luciferase and the various oxidoreductases from Vibrio harveyi, a bright mutant of...
Gespeichert in:
| Veröffentlicht in: | Journal of bioluminescence and chemiluminescence 1990-07, Vol.5 (3), p.187-192 |
|---|---|
| Hauptverfasser: | , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 192 |
|---|---|
| container_issue | 3 |
| container_start_page | 187 |
| container_title | Journal of bioluminescence and chemiluminescence |
| container_volume | 5 |
| creator | Lavi, Jukka T. Raunio, Raimo P. Stahlberg, Tom H. |
| description | A modified purification method for bacterial luciferases and NAD(P)H:FMN oxidoreductases is described which uses FMN-Sepharose alone or coupled to DEAE ion exchange chromatography for the simultaneous purification of luciferase and the various oxidoreductases from Vibrio harveyi, a bright mutant of Vibrio harveyi, Vibrio fischeri, and Photobacterium phosphoreum. This purification method is compared with DEAE-Sepharose Cl 6B fractionations from these organisms. Both methods allow the separation of oxidoreductases specific for either NADH or NADPH. The use of FMN-Sepharose coupled to DEAE-Sepharose fractionation allows the isolation of highly purified enzymes. Lacking interfering factors, these are very suitable for various analytical applications based on bacterial bioluminescence enzymes. The partially purified enzymes from the affinity column have higher specific activities than those obtained using DEAE-Sepharose. |
| doi_str_mv | 10.1002/bio.1170050308 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_80064702</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>80064702</sourcerecordid><originalsourceid>FETCH-LOGICAL-c328t-e38d5b6af3404f8cf6dc4b9a8d0be07b482ccdd4d05d32aa1f274d44f91fa3313</originalsourceid><addsrcrecordid>eNpFkMtv1DAQhy0EKtvClRuSTwgOWcaPJA63VaFdRNlWVRFHa-KHMGTXqZ1I3f71GO2qnEaa32NGHyFvGCwZAP_Yh7hkrAWoQYB6RhYMuq5ivGXPyQKUkpXouuYlOc35NwA0XS1OyAnnHCRrFuRx5X3YhWlPxzkFHwxOIe5o9LRHM7kUcKDDbIJ3CbOjuLN0s_r8_ubD-tPF9w2ND8HG5OxspiJn2u9pWVfZjb8wxRLwMZUQDvupVA8Ux3E43sivyAuPQ3avj_OM_Lj4cne-rq6uL7-er64qI7iaKieUrfsGvZAgvTK-sUb2HSoLvYO2l4obY620UFvBEZnnrbRS-o55FIKJM_Lu0DumeD-7POltyMYNA-5cnLNWhYpsgRfj8mA05fWcnNdjCltMe81A_4OtC2z9H3YJvD02z_3W2Sf7kW7Rq4Me8uQenmRMf3TTirbWPzeX-tsth3YtQXPxFwGuiyE</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>80064702</pqid></control><display><type>article</type><title>Affinity purification of bacterial luciferase and NAD(P)H:FMN oxidoreductases by FMN-sepharose for analytical applications</title><source>MEDLINE</source><source>Wiley Online Library All Journals</source><creator>Lavi, Jukka T. ; Raunio, Raimo P. ; Stahlberg, Tom H.</creator><creatorcontrib>Lavi, Jukka T. ; Raunio, Raimo P. ; Stahlberg, Tom H.</creatorcontrib><description>A modified purification method for bacterial luciferases and NAD(P)H:FMN oxidoreductases is described which uses FMN-Sepharose alone or coupled to DEAE ion exchange chromatography for the simultaneous purification of luciferase and the various oxidoreductases from Vibrio harveyi, a bright mutant of Vibrio harveyi, Vibrio fischeri, and Photobacterium phosphoreum. This purification method is compared with DEAE-Sepharose Cl 6B fractionations from these organisms. Both methods allow the separation of oxidoreductases specific for either NADH or NADPH. The use of FMN-Sepharose coupled to DEAE-Sepharose fractionation allows the isolation of highly purified enzymes. Lacking interfering factors, these are very suitable for various analytical applications based on bacterial bioluminescence enzymes. The partially purified enzymes from the affinity column have higher specific activities than those obtained using DEAE-Sepharose.</description><identifier>ISSN: 0884-3996</identifier><identifier>EISSN: 1099-1271</identifier><identifier>DOI: 10.1002/bio.1170050308</identifier><identifier>PMID: 2220416</identifier><language>eng</language><publisher>Chichester, UK: John Wiley & Sons, Ltd</publisher><subject>affinity purification ; Bacterial luciferase ; Chromatography, Affinity - methods ; Chromatography, Ion Exchange ; FMN Reductase ; Luciferases - isolation & purification ; Luminescent Measurements ; NAD ; NADH, NADPH Oxidoreductases - isolation & purification ; NADP ; oxidoreductases ; Photobacterium - enzymology ; Substrate Specificity ; Vibrio - enzymology</subject><ispartof>Journal of bioluminescence and chemiluminescence, 1990-07, Vol.5 (3), p.187-192</ispartof><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c328t-e38d5b6af3404f8cf6dc4b9a8d0be07b482ccdd4d05d32aa1f274d44f91fa3313</citedby><cites>FETCH-LOGICAL-c328t-e38d5b6af3404f8cf6dc4b9a8d0be07b482ccdd4d05d32aa1f274d44f91fa3313</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27923,27924</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/2220416$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Lavi, Jukka T.</creatorcontrib><creatorcontrib>Raunio, Raimo P.</creatorcontrib><creatorcontrib>Stahlberg, Tom H.</creatorcontrib><title>Affinity purification of bacterial luciferase and NAD(P)H:FMN oxidoreductases by FMN-sepharose for analytical applications</title><title>Journal of bioluminescence and chemiluminescence</title><addtitle>J. Biolumin. Chemilumin</addtitle><description>A modified purification method for bacterial luciferases and NAD(P)H:FMN oxidoreductases is described which uses FMN-Sepharose alone or coupled to DEAE ion exchange chromatography for the simultaneous purification of luciferase and the various oxidoreductases from Vibrio harveyi, a bright mutant of Vibrio harveyi, Vibrio fischeri, and Photobacterium phosphoreum. This purification method is compared with DEAE-Sepharose Cl 6B fractionations from these organisms. Both methods allow the separation of oxidoreductases specific for either NADH or NADPH. The use of FMN-Sepharose coupled to DEAE-Sepharose fractionation allows the isolation of highly purified enzymes. Lacking interfering factors, these are very suitable for various analytical applications based on bacterial bioluminescence enzymes. The partially purified enzymes from the affinity column have higher specific activities than those obtained using DEAE-Sepharose.</description><subject>affinity purification</subject><subject>Bacterial luciferase</subject><subject>Chromatography, Affinity - methods</subject><subject>Chromatography, Ion Exchange</subject><subject>FMN Reductase</subject><subject>Luciferases - isolation & purification</subject><subject>Luminescent Measurements</subject><subject>NAD</subject><subject>NADH, NADPH Oxidoreductases - isolation & purification</subject><subject>NADP</subject><subject>oxidoreductases</subject><subject>Photobacterium - enzymology</subject><subject>Substrate Specificity</subject><subject>Vibrio - enzymology</subject><issn>0884-3996</issn><issn>1099-1271</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1990</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpFkMtv1DAQhy0EKtvClRuSTwgOWcaPJA63VaFdRNlWVRFHa-KHMGTXqZ1I3f71GO2qnEaa32NGHyFvGCwZAP_Yh7hkrAWoQYB6RhYMuq5ivGXPyQKUkpXouuYlOc35NwA0XS1OyAnnHCRrFuRx5X3YhWlPxzkFHwxOIe5o9LRHM7kUcKDDbIJ3CbOjuLN0s_r8_ubD-tPF9w2ND8HG5OxspiJn2u9pWVfZjb8wxRLwMZUQDvupVA8Ux3E43sivyAuPQ3avj_OM_Lj4cne-rq6uL7-er64qI7iaKieUrfsGvZAgvTK-sUb2HSoLvYO2l4obY620UFvBEZnnrbRS-o55FIKJM_Lu0DumeD-7POltyMYNA-5cnLNWhYpsgRfj8mA05fWcnNdjCltMe81A_4OtC2z9H3YJvD02z_3W2Sf7kW7Rq4Me8uQenmRMf3TTirbWPzeX-tsth3YtQXPxFwGuiyE</recordid><startdate>19900701</startdate><enddate>19900701</enddate><creator>Lavi, Jukka T.</creator><creator>Raunio, Raimo P.</creator><creator>Stahlberg, Tom H.</creator><general>John Wiley & Sons, Ltd</general><scope>BSCLL</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>7X8</scope></search><sort><creationdate>19900701</creationdate><title>Affinity purification of bacterial luciferase and NAD(P)H:FMN oxidoreductases by FMN-sepharose for analytical applications</title><author>Lavi, Jukka T. ; Raunio, Raimo P. ; Stahlberg, Tom H.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c328t-e38d5b6af3404f8cf6dc4b9a8d0be07b482ccdd4d05d32aa1f274d44f91fa3313</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1990</creationdate><topic>affinity purification</topic><topic>Bacterial luciferase</topic><topic>Chromatography, Affinity - methods</topic><topic>Chromatography, Ion Exchange</topic><topic>FMN Reductase</topic><topic>Luciferases - isolation & purification</topic><topic>Luminescent Measurements</topic><topic>NAD</topic><topic>NADH, NADPH Oxidoreductases - isolation & purification</topic><topic>NADP</topic><topic>oxidoreductases</topic><topic>Photobacterium - enzymology</topic><topic>Substrate Specificity</topic><topic>Vibrio - enzymology</topic><toplevel>online_resources</toplevel><creatorcontrib>Lavi, Jukka T.</creatorcontrib><creatorcontrib>Raunio, Raimo P.</creatorcontrib><creatorcontrib>Stahlberg, Tom H.</creatorcontrib><collection>Istex</collection><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 bioluminescence and chemiluminescence</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Lavi, Jukka T.</au><au>Raunio, Raimo P.</au><au>Stahlberg, Tom H.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Affinity purification of bacterial luciferase and NAD(P)H:FMN oxidoreductases by FMN-sepharose for analytical applications</atitle><jtitle>Journal of bioluminescence and chemiluminescence</jtitle><addtitle>J. Biolumin. Chemilumin</addtitle><date>1990-07-01</date><risdate>1990</risdate><volume>5</volume><issue>3</issue><spage>187</spage><epage>192</epage><pages>187-192</pages><issn>0884-3996</issn><eissn>1099-1271</eissn><abstract>A modified purification method for bacterial luciferases and NAD(P)H:FMN oxidoreductases is described which uses FMN-Sepharose alone or coupled to DEAE ion exchange chromatography for the simultaneous purification of luciferase and the various oxidoreductases from Vibrio harveyi, a bright mutant of Vibrio harveyi, Vibrio fischeri, and Photobacterium phosphoreum. This purification method is compared with DEAE-Sepharose Cl 6B fractionations from these organisms. Both methods allow the separation of oxidoreductases specific for either NADH or NADPH. The use of FMN-Sepharose coupled to DEAE-Sepharose fractionation allows the isolation of highly purified enzymes. Lacking interfering factors, these are very suitable for various analytical applications based on bacterial bioluminescence enzymes. The partially purified enzymes from the affinity column have higher specific activities than those obtained using DEAE-Sepharose.</abstract><cop>Chichester, UK</cop><pub>John Wiley & Sons, Ltd</pub><pmid>2220416</pmid><doi>10.1002/bio.1170050308</doi><tpages>6</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0884-3996 |
| ispartof | Journal of bioluminescence and chemiluminescence, 1990-07, Vol.5 (3), p.187-192 |
| issn | 0884-3996 1099-1271 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_80064702 |
| source | MEDLINE; Wiley Online Library All Journals |
| subjects | affinity purification Bacterial luciferase Chromatography, Affinity - methods Chromatography, Ion Exchange FMN Reductase Luciferases - isolation & purification Luminescent Measurements NAD NADH, NADPH Oxidoreductases - isolation & purification NADP oxidoreductases Photobacterium - enzymology Substrate Specificity Vibrio - enzymology |
| title | Affinity purification of bacterial luciferase and NAD(P)H:FMN oxidoreductases by FMN-sepharose for analytical applications |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-11T22%3A45%3A16IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Affinity%20purification%20of%20bacterial%20luciferase%20and%20NAD(P)H:FMN%20oxidoreductases%20by%20FMN-sepharose%20for%20analytical%20applications&rft.jtitle=Journal%20of%20bioluminescence%20and%20chemiluminescence&rft.au=Lavi,%20Jukka%20T.&rft.date=1990-07-01&rft.volume=5&rft.issue=3&rft.spage=187&rft.epage=192&rft.pages=187-192&rft.issn=0884-3996&rft.eissn=1099-1271&rft_id=info:doi/10.1002/bio.1170050308&rft_dat=%3Cproquest_cross%3E80064702%3C/proquest_cross%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=80064702&rft_id=info:pmid/2220416&rfr_iscdi=true |