Oxygenation Cascade in Conversion of n-Alkanes to α,ω-Dioic Acids Catalyzed by Cytochrome P450 52A3
Purified recombinant cytochrome P450 52A3 and the corresponding NADPH-cytochrome P450 reductase from the alkane-assimilating yeast Candida maltosa were reconstituted into an active alkane monooxygenase system. Besides the primary product, 1-hexadecanol, the conversion of hexadecane yielded up to fiv...
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| Veröffentlicht in: | The Journal of biological chemistry 1998-12, Vol.273 (49), p.32528-32534 |
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| creator | Scheller, Ulrich Zimmer, Thomas Becher, Dörte Schauer, Frieder Schunck, Wolf-Hagen |
| description | Purified recombinant cytochrome P450 52A3 and the corresponding NADPH-cytochrome P450 reductase from the alkane-assimilating yeast Candida maltosa were reconstituted into an active alkane monooxygenase system. Besides the primary product, 1-hexadecanol, the conversion of hexadecane yielded up to five additional metabolites, which were identified by gas chromatography-electron impact mass spectrometry as hexadecanal, hexadecanoic acid, 1,16-hexadecanediol, 16-hydroxyhexadecanoic acid, and 1,16-hexadecanedioic acid. As shown by substrate binding studies, the final product 1,16-hexadecanedioic acid acts as a competitive inhibitor of n-alkane binding and may be important for the metabolic regulation of the P450 activity. Kinetic studies of the individual sequential reactions revealed highVmax values for the conversion of hexadecane, 1-hexadecanol, and hexadecanal (27, 23, and 69 min−1, respectively), whereas the oxidation of hexadecanoic acid, 1,16-hexadecanediol, and 16-hydroxyhexadecanoic acid occurred at significantly lower rates (9, 9, and 5 min−1, respectively). 1-Hexadecanol was found to be the main branch point between mono- and diterminal oxidation. Taken together with data on the incorporation of 18O2-derived oxygen into the hexadecane oxidation products, the present study demonstrates that a single P450 form is able to efficiently catalyze a cascade of sequential mono- and diterminal monooxygenation reactions fromn-alkanes to α,ω-dioic acids with high regioselectivity. |
| doi_str_mv | 10.1074/jbc.273.49.32528 |
| format | Article |
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Besides the primary product, 1-hexadecanol, the conversion of hexadecane yielded up to five additional metabolites, which were identified by gas chromatography-electron impact mass spectrometry as hexadecanal, hexadecanoic acid, 1,16-hexadecanediol, 16-hydroxyhexadecanoic acid, and 1,16-hexadecanedioic acid. As shown by substrate binding studies, the final product 1,16-hexadecanedioic acid acts as a competitive inhibitor of n-alkane binding and may be important for the metabolic regulation of the P450 activity. Kinetic studies of the individual sequential reactions revealed highVmax values for the conversion of hexadecane, 1-hexadecanol, and hexadecanal (27, 23, and 69 min−1, respectively), whereas the oxidation of hexadecanoic acid, 1,16-hexadecanediol, and 16-hydroxyhexadecanoic acid occurred at significantly lower rates (9, 9, and 5 min−1, respectively). 1-Hexadecanol was found to be the main branch point between mono- and diterminal oxidation. Taken together with data on the incorporation of 18O2-derived oxygen into the hexadecane oxidation products, the present study demonstrates that a single P450 form is able to efficiently catalyze a cascade of sequential mono- and diterminal monooxygenation reactions fromn-alkanes to α,ω-dioic acids with high regioselectivity.</description><identifier>ISSN: 0021-9258</identifier><identifier>EISSN: 1083-351X</identifier><identifier>DOI: 10.1074/jbc.273.49.32528</identifier><identifier>PMID: 9829987</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>1-HEXADECANOL ; ACIDE ORGANIQUE ; ACIDOS ORGANICOS ; ACTIVIDAD ENZIMATICA ; ACTIVITE ENZYMATIQUE ; ALCOHOLES ; ALCOHOLES GRASOS ; ALCOHOLS ; ALCOOL ; ALCOOL GRAS ; ALKANE MONOOXYGENASE ; Alkanes - metabolism ; CANDIDA MALTOSA ; CARBOXYLIC ACIDS ; Catalysis ; CHEMICAL STRUCTURE ; CITOCROMO P450 ; Cytochrome P-450 Enzyme System - metabolism ; CYTOCHROME P450 ; Dicarboxylic Acids - metabolism ; ENZYMIC ACTIVITY ; ESTRUCTURA QUIMICA ; FATTY ALCOHOLS ; Fungal Proteins ; Gas Chromatography-Mass Spectrometry ; HEXADECANE ; HIDROCARBUROS ; HYDROCARBONS ; HYDROCARBURE ; Kinetics ; METABOLITE ; METABOLITES ; METABOLITOS ; Mixed Function Oxygenases ; ORGANIC ACIDS ; OXIDACION ; OXIDATION ; OXIDOREDUCTASES ; OXIDORREDUCTASAS ; OXYDATION ; OXYDOREDUCTASE ; Oxygen - metabolism ; PROTEINAS RECOMBINANTES ; PROTEINE RECOMBINANTE ; RECOMBINANT PROTEINS ; STRUCTURE CHIMIQUE ; UNSPECIFIC MONOOXYGENASE</subject><ispartof>The Journal of biological chemistry, 1998-12, Vol.273 (49), p.32528-32534</ispartof><rights>1998 © 1998 ASBMB. Currently published by Elsevier Inc; originally published by American Society for Biochemistry and Molecular Biology.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c411t-90cb43fe16ef773f569cbca572f367d3ca930bb73e10897fc7297309209875a13</citedby><cites>FETCH-LOGICAL-c411t-90cb43fe16ef773f569cbca572f367d3ca930bb73e10897fc7297309209875a13</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/9829987$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Scheller, Ulrich</creatorcontrib><creatorcontrib>Zimmer, Thomas</creatorcontrib><creatorcontrib>Becher, Dörte</creatorcontrib><creatorcontrib>Schauer, Frieder</creatorcontrib><creatorcontrib>Schunck, Wolf-Hagen</creatorcontrib><title>Oxygenation Cascade in Conversion of n-Alkanes to α,ω-Dioic Acids Catalyzed by Cytochrome P450 52A3</title><title>The Journal of biological chemistry</title><addtitle>J Biol Chem</addtitle><description>Purified recombinant cytochrome P450 52A3 and the corresponding NADPH-cytochrome P450 reductase from the alkane-assimilating yeast Candida maltosa were reconstituted into an active alkane monooxygenase system. Besides the primary product, 1-hexadecanol, the conversion of hexadecane yielded up to five additional metabolites, which were identified by gas chromatography-electron impact mass spectrometry as hexadecanal, hexadecanoic acid, 1,16-hexadecanediol, 16-hydroxyhexadecanoic acid, and 1,16-hexadecanedioic acid. As shown by substrate binding studies, the final product 1,16-hexadecanedioic acid acts as a competitive inhibitor of n-alkane binding and may be important for the metabolic regulation of the P450 activity. Kinetic studies of the individual sequential reactions revealed highVmax values for the conversion of hexadecane, 1-hexadecanol, and hexadecanal (27, 23, and 69 min−1, respectively), whereas the oxidation of hexadecanoic acid, 1,16-hexadecanediol, and 16-hydroxyhexadecanoic acid occurred at significantly lower rates (9, 9, and 5 min−1, respectively). 1-Hexadecanol was found to be the main branch point between mono- and diterminal oxidation. Taken together with data on the incorporation of 18O2-derived oxygen into the hexadecane oxidation products, the present study demonstrates that a single P450 form is able to efficiently catalyze a cascade of sequential mono- and diterminal monooxygenation reactions fromn-alkanes to α,ω-dioic acids with high regioselectivity.</description><subject>1-HEXADECANOL</subject><subject>ACIDE ORGANIQUE</subject><subject>ACIDOS ORGANICOS</subject><subject>ACTIVIDAD ENZIMATICA</subject><subject>ACTIVITE ENZYMATIQUE</subject><subject>ALCOHOLES</subject><subject>ALCOHOLES GRASOS</subject><subject>ALCOHOLS</subject><subject>ALCOOL</subject><subject>ALCOOL GRAS</subject><subject>ALKANE MONOOXYGENASE</subject><subject>Alkanes - metabolism</subject><subject>CANDIDA MALTOSA</subject><subject>CARBOXYLIC ACIDS</subject><subject>Catalysis</subject><subject>CHEMICAL STRUCTURE</subject><subject>CITOCROMO P450</subject><subject>Cytochrome P-450 Enzyme System - metabolism</subject><subject>CYTOCHROME P450</subject><subject>Dicarboxylic Acids - metabolism</subject><subject>ENZYMIC ACTIVITY</subject><subject>ESTRUCTURA QUIMICA</subject><subject>FATTY ALCOHOLS</subject><subject>Fungal Proteins</subject><subject>Gas Chromatography-Mass Spectrometry</subject><subject>HEXADECANE</subject><subject>HIDROCARBUROS</subject><subject>HYDROCARBONS</subject><subject>HYDROCARBURE</subject><subject>Kinetics</subject><subject>METABOLITE</subject><subject>METABOLITES</subject><subject>METABOLITOS</subject><subject>Mixed Function Oxygenases</subject><subject>ORGANIC ACIDS</subject><subject>OXIDACION</subject><subject>OXIDATION</subject><subject>OXIDOREDUCTASES</subject><subject>OXIDORREDUCTASAS</subject><subject>OXYDATION</subject><subject>OXYDOREDUCTASE</subject><subject>Oxygen - metabolism</subject><subject>PROTEINAS RECOMBINANTES</subject><subject>PROTEINE RECOMBINANTE</subject><subject>RECOMBINANT PROTEINS</subject><subject>STRUCTURE CHIMIQUE</subject><subject>UNSPECIFIC MONOOXYGENASE</subject><issn>0021-9258</issn><issn>1083-351X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1998</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp1kM1O3DAQx62qiC60916QfOqpWfwRxzG31fIpIYHUIvVmOc4YTLMxtbOo4Q14HF4EXqledgWnzmVGM_Ofjx9CXymZUiLL_dvGTpnk01JNOROs_oAmlNS84IL--ogmhDBaKCbqT2gnpVuSrVR0G22rmilVywmCi7_jNfRm8KHHc5OsaQH7HIb-HmJaZYPDfTHrfpseEh4Cfn76_vJYHPrgLZ5Z36asG0w3PkCLmxHPxyHYmxgWgC9LQbBgM_4ZbTnTJfiy8bvo6vjo5_y0OL84OZvPzgtbUjoUitim5A5oBU5K7kSlbGONkMzxSrbcGsVJ00gO-UklnZVMSU4UI_kXYSjfRd_Wc-9i-LOENOiFTxa6Lt8elklLQmTNRZkbybrRxpBSBKfvol-YOGpK9IqszmR1JqtLpV_JZsneZvayWUD7JtigfK87E7S5jj7pqx9UKZV3VtXqtoN1HTKAew9RJ-uht9D6CHbQbfD_X_4PYqyP8g</recordid><startdate>19981204</startdate><enddate>19981204</enddate><creator>Scheller, Ulrich</creator><creator>Zimmer, Thomas</creator><creator>Becher, Dörte</creator><creator>Schauer, Frieder</creator><creator>Schunck, Wolf-Hagen</creator><general>Elsevier Inc</general><scope>6I.</scope><scope>AAFTH</scope><scope>FBQ</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>19981204</creationdate><title>Oxygenation Cascade in Conversion of n-Alkanes to α,ω-Dioic Acids Catalyzed by Cytochrome P450 52A3</title><author>Scheller, Ulrich ; Zimmer, Thomas ; Becher, Dörte ; Schauer, Frieder ; Schunck, Wolf-Hagen</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c411t-90cb43fe16ef773f569cbca572f367d3ca930bb73e10897fc7297309209875a13</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1998</creationdate><topic>1-HEXADECANOL</topic><topic>ACIDE ORGANIQUE</topic><topic>ACIDOS ORGANICOS</topic><topic>ACTIVIDAD ENZIMATICA</topic><topic>ACTIVITE ENZYMATIQUE</topic><topic>ALCOHOLES</topic><topic>ALCOHOLES GRASOS</topic><topic>ALCOHOLS</topic><topic>ALCOOL</topic><topic>ALCOOL GRAS</topic><topic>ALKANE MONOOXYGENASE</topic><topic>Alkanes - metabolism</topic><topic>CANDIDA MALTOSA</topic><topic>CARBOXYLIC ACIDS</topic><topic>Catalysis</topic><topic>CHEMICAL STRUCTURE</topic><topic>CITOCROMO P450</topic><topic>Cytochrome P-450 Enzyme System - metabolism</topic><topic>CYTOCHROME P450</topic><topic>Dicarboxylic Acids - metabolism</topic><topic>ENZYMIC ACTIVITY</topic><topic>ESTRUCTURA QUIMICA</topic><topic>FATTY ALCOHOLS</topic><topic>Fungal Proteins</topic><topic>Gas Chromatography-Mass Spectrometry</topic><topic>HEXADECANE</topic><topic>HIDROCARBUROS</topic><topic>HYDROCARBONS</topic><topic>HYDROCARBURE</topic><topic>Kinetics</topic><topic>METABOLITE</topic><topic>METABOLITES</topic><topic>METABOLITOS</topic><topic>Mixed Function Oxygenases</topic><topic>ORGANIC ACIDS</topic><topic>OXIDACION</topic><topic>OXIDATION</topic><topic>OXIDOREDUCTASES</topic><topic>OXIDORREDUCTASAS</topic><topic>OXYDATION</topic><topic>OXYDOREDUCTASE</topic><topic>Oxygen - metabolism</topic><topic>PROTEINAS RECOMBINANTES</topic><topic>PROTEINE RECOMBINANTE</topic><topic>RECOMBINANT PROTEINS</topic><topic>STRUCTURE CHIMIQUE</topic><topic>UNSPECIFIC MONOOXYGENASE</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Scheller, Ulrich</creatorcontrib><creatorcontrib>Zimmer, Thomas</creatorcontrib><creatorcontrib>Becher, Dörte</creatorcontrib><creatorcontrib>Schauer, Frieder</creatorcontrib><creatorcontrib>Schunck, Wolf-Hagen</creatorcontrib><collection>ScienceDirect Open Access Titles</collection><collection>Elsevier:ScienceDirect:Open Access</collection><collection>AGRIS</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>The Journal of biological chemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Scheller, Ulrich</au><au>Zimmer, Thomas</au><au>Becher, Dörte</au><au>Schauer, Frieder</au><au>Schunck, Wolf-Hagen</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Oxygenation Cascade in Conversion of n-Alkanes to α,ω-Dioic Acids Catalyzed by Cytochrome P450 52A3</atitle><jtitle>The Journal of biological chemistry</jtitle><addtitle>J Biol Chem</addtitle><date>1998-12-04</date><risdate>1998</risdate><volume>273</volume><issue>49</issue><spage>32528</spage><epage>32534</epage><pages>32528-32534</pages><issn>0021-9258</issn><eissn>1083-351X</eissn><abstract>Purified recombinant cytochrome P450 52A3 and the corresponding NADPH-cytochrome P450 reductase from the alkane-assimilating yeast Candida maltosa were reconstituted into an active alkane monooxygenase system. Besides the primary product, 1-hexadecanol, the conversion of hexadecane yielded up to five additional metabolites, which were identified by gas chromatography-electron impact mass spectrometry as hexadecanal, hexadecanoic acid, 1,16-hexadecanediol, 16-hydroxyhexadecanoic acid, and 1,16-hexadecanedioic acid. As shown by substrate binding studies, the final product 1,16-hexadecanedioic acid acts as a competitive inhibitor of n-alkane binding and may be important for the metabolic regulation of the P450 activity. Kinetic studies of the individual sequential reactions revealed highVmax values for the conversion of hexadecane, 1-hexadecanol, and hexadecanal (27, 23, and 69 min−1, respectively), whereas the oxidation of hexadecanoic acid, 1,16-hexadecanediol, and 16-hydroxyhexadecanoic acid occurred at significantly lower rates (9, 9, and 5 min−1, respectively). 1-Hexadecanol was found to be the main branch point between mono- and diterminal oxidation. Taken together with data on the incorporation of 18O2-derived oxygen into the hexadecane oxidation products, the present study demonstrates that a single P450 form is able to efficiently catalyze a cascade of sequential mono- and diterminal monooxygenation reactions fromn-alkanes to α,ω-dioic acids with high regioselectivity.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>9829987</pmid><doi>10.1074/jbc.273.49.32528</doi><tpages>7</tpages><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0021-9258 |
| ispartof | The Journal of biological chemistry, 1998-12, Vol.273 (49), p.32528-32534 |
| issn | 0021-9258 1083-351X |
| language | eng |
| recordid | cdi_proquest_miscellaneous_70078354 |
| source | MEDLINE; EZB-FREE-00999 freely available EZB journals; Alma/SFX Local Collection |
| subjects | 1-HEXADECANOL ACIDE ORGANIQUE ACIDOS ORGANICOS ACTIVIDAD ENZIMATICA ACTIVITE ENZYMATIQUE ALCOHOLES ALCOHOLES GRASOS ALCOHOLS ALCOOL ALCOOL GRAS ALKANE MONOOXYGENASE Alkanes - metabolism CANDIDA MALTOSA CARBOXYLIC ACIDS Catalysis CHEMICAL STRUCTURE CITOCROMO P450 Cytochrome P-450 Enzyme System - metabolism CYTOCHROME P450 Dicarboxylic Acids - metabolism ENZYMIC ACTIVITY ESTRUCTURA QUIMICA FATTY ALCOHOLS Fungal Proteins Gas Chromatography-Mass Spectrometry HEXADECANE HIDROCARBUROS HYDROCARBONS HYDROCARBURE Kinetics METABOLITE METABOLITES METABOLITOS Mixed Function Oxygenases ORGANIC ACIDS OXIDACION OXIDATION OXIDOREDUCTASES OXIDORREDUCTASAS OXYDATION OXYDOREDUCTASE Oxygen - metabolism PROTEINAS RECOMBINANTES PROTEINE RECOMBINANTE RECOMBINANT PROTEINS STRUCTURE CHIMIQUE UNSPECIFIC MONOOXYGENASE |
| title | Oxygenation Cascade in Conversion of n-Alkanes to α,ω-Dioic Acids Catalyzed by Cytochrome P450 52A3 |
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