Purification and Characterization of a Novel NAD(P)+-Farnesol Dehydrogenase from Polygonum minus Leaves
Identifieur interne : 001E12 ( Ncbi/Merge ); précédent : 001E11; suivant : 001E13Purification and Characterization of a Novel NAD(P)+-Farnesol Dehydrogenase from Polygonum minus Leaves
Auteurs : Nor-Ain-Shahajar Ahmad-Sohdi [Malaisie] ; Ahmad-Faris Seman-Kamarulzaman [Malaisie] ; Zeti-Azura Mohamed-Hussein [Malaisie] ; Maizom Hassan [Malaisie]Source :
- PLoS ONE [ 1932-6203 ] ; 2015.
Abstract
Juvenile hormones have attracted attention as safe and selective targets for the design and development of environmentally friendly and biorational insecticides. In the juvenile hormone III biosynthetic pathway, the enzyme farnesol dehydrogenase catalyzes the oxidation of farnesol to farnesal. In this study, farnesol dehydrogenase was extracted from
Url:
DOI: 10.1371/journal.pone.0143310
PubMed: 26600471
PubMed Central: 4657912
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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Purification and Characterization of a Novel NAD(P)<sup>+</sup>-Farnesol Dehydrogenase from <italic>Polygonum minus</italic> Leaves</title><author><name sortKey="Ahmad Sohdi, Nor Ain Shahajar" sort="Ahmad Sohdi, Nor Ain Shahajar" uniqKey="Ahmad Sohdi N" first="Nor-Ain-Shahajar" last="Ahmad-Sohdi">Nor-Ain-Shahajar Ahmad-Sohdi</name><affiliation wicri:level="1"><nlm:aff id="aff001"><addr-line>Institute of Systems Biology, Universiti Kebangsaan Malaysia (UKM), 43600 UKM, Bangi, Selangor, Malaysia</addr-line></nlm:aff><country xml:lang="fr">Malaisie</country><wicri:regionArea>Institute of Systems Biology, Universiti Kebangsaan Malaysia (UKM), 43600 UKM, Bangi, Selangor</wicri:regionArea><wicri:noRegion>Selangor</wicri:noRegion></affiliation></author><author><name sortKey="Seman Kamarulzaman, Ahmad Faris" sort="Seman Kamarulzaman, Ahmad Faris" uniqKey="Seman Kamarulzaman A" first="Ahmad-Faris" last="Seman-Kamarulzaman">Ahmad-Faris Seman-Kamarulzaman</name><affiliation wicri:level="1"><nlm:aff id="aff001"><addr-line>Institute of Systems Biology, Universiti Kebangsaan Malaysia (UKM), 43600 UKM, Bangi, Selangor, Malaysia</addr-line></nlm:aff><country xml:lang="fr">Malaisie</country><wicri:regionArea>Institute of Systems Biology, Universiti Kebangsaan Malaysia (UKM), 43600 UKM, Bangi, Selangor</wicri:regionArea><wicri:noRegion>Selangor</wicri:noRegion></affiliation></author><author><name sortKey="Mohamed Hussein, Zeti Azura" sort="Mohamed Hussein, Zeti Azura" uniqKey="Mohamed Hussein Z" first="Zeti-Azura" last="Mohamed-Hussein">Zeti-Azura Mohamed-Hussein</name><affiliation wicri:level="1"><nlm:aff id="aff001"><addr-line>Institute of Systems Biology, Universiti Kebangsaan Malaysia (UKM), 43600 UKM, Bangi, Selangor, Malaysia</addr-line></nlm:aff><country xml:lang="fr">Malaisie</country><wicri:regionArea>Institute of Systems Biology, Universiti Kebangsaan Malaysia (UKM), 43600 UKM, Bangi, Selangor</wicri:regionArea><wicri:noRegion>Selangor</wicri:noRegion></affiliation><affiliation wicri:level="1"><nlm:aff id="aff002"><addr-line>School of Biosciences and Biotechnology, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, 43600 UKM, Bangi, Selangor, Malaysia</addr-line></nlm:aff><country xml:lang="fr">Malaisie</country><wicri:regionArea>School of Biosciences and Biotechnology, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, 43600 UKM, Bangi, Selangor</wicri:regionArea><wicri:noRegion>Selangor</wicri:noRegion></affiliation></author><author><name sortKey="Hassan, Maizom" sort="Hassan, Maizom" uniqKey="Hassan M" first="Maizom" last="Hassan">Maizom Hassan</name><affiliation wicri:level="1"><nlm:aff id="aff001"><addr-line>Institute of Systems Biology, Universiti Kebangsaan Malaysia (UKM), 43600 UKM, Bangi, Selangor, Malaysia</addr-line></nlm:aff><country xml:lang="fr">Malaisie</country><wicri:regionArea>Institute of Systems Biology, Universiti Kebangsaan Malaysia (UKM), 43600 UKM, Bangi, Selangor</wicri:regionArea><wicri:noRegion>Selangor</wicri:noRegion></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">26600471</idno><idno type="pmc">4657912</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4657912</idno><idno type="RBID">PMC:4657912</idno><idno type="doi">10.1371/journal.pone.0143310</idno><date when="2015">2015</date><idno type="wicri:Area/Pmc/Corpus">000202</idno><idno type="wicri:Area/Pmc/Curation">000202</idno><idno type="wicri:Area/Pmc/Checkpoint">000371</idno><idno type="wicri:Area/Ncbi/Merge">001E12</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Purification and Characterization of a Novel NAD(P)<sup>+</sup>-Farnesol Dehydrogenase from <italic>Polygonum minus</italic> Leaves</title><author><name sortKey="Ahmad Sohdi, Nor Ain Shahajar" sort="Ahmad Sohdi, Nor Ain Shahajar" uniqKey="Ahmad Sohdi N" first="Nor-Ain-Shahajar" last="Ahmad-Sohdi">Nor-Ain-Shahajar Ahmad-Sohdi</name><affiliation wicri:level="1"><nlm:aff id="aff001"><addr-line>Institute of Systems Biology, Universiti Kebangsaan Malaysia (UKM), 43600 UKM, Bangi, Selangor, Malaysia</addr-line></nlm:aff><country xml:lang="fr">Malaisie</country><wicri:regionArea>Institute of Systems Biology, Universiti Kebangsaan Malaysia (UKM), 43600 UKM, Bangi, Selangor</wicri:regionArea><wicri:noRegion>Selangor</wicri:noRegion></affiliation></author><author><name sortKey="Seman Kamarulzaman, Ahmad Faris" sort="Seman Kamarulzaman, Ahmad Faris" uniqKey="Seman Kamarulzaman A" first="Ahmad-Faris" last="Seman-Kamarulzaman">Ahmad-Faris Seman-Kamarulzaman</name><affiliation wicri:level="1"><nlm:aff id="aff001"><addr-line>Institute of Systems Biology, Universiti Kebangsaan Malaysia (UKM), 43600 UKM, Bangi, Selangor, Malaysia</addr-line></nlm:aff><country xml:lang="fr">Malaisie</country><wicri:regionArea>Institute of Systems Biology, Universiti Kebangsaan Malaysia (UKM), 43600 UKM, Bangi, Selangor</wicri:regionArea><wicri:noRegion>Selangor</wicri:noRegion></affiliation></author><author><name sortKey="Mohamed Hussein, Zeti Azura" sort="Mohamed Hussein, Zeti Azura" uniqKey="Mohamed Hussein Z" first="Zeti-Azura" last="Mohamed-Hussein">Zeti-Azura Mohamed-Hussein</name><affiliation wicri:level="1"><nlm:aff id="aff001"><addr-line>Institute of Systems Biology, Universiti Kebangsaan Malaysia (UKM), 43600 UKM, Bangi, Selangor, Malaysia</addr-line></nlm:aff><country xml:lang="fr">Malaisie</country><wicri:regionArea>Institute of Systems Biology, Universiti Kebangsaan Malaysia (UKM), 43600 UKM, Bangi, Selangor</wicri:regionArea><wicri:noRegion>Selangor</wicri:noRegion></affiliation><affiliation wicri:level="1"><nlm:aff id="aff002"><addr-line>School of Biosciences and Biotechnology, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, 43600 UKM, Bangi, Selangor, Malaysia</addr-line></nlm:aff><country xml:lang="fr">Malaisie</country><wicri:regionArea>School of Biosciences and Biotechnology, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, 43600 UKM, Bangi, Selangor</wicri:regionArea><wicri:noRegion>Selangor</wicri:noRegion></affiliation></author><author><name sortKey="Hassan, Maizom" sort="Hassan, Maizom" uniqKey="Hassan M" first="Maizom" last="Hassan">Maizom Hassan</name><affiliation wicri:level="1"><nlm:aff id="aff001"><addr-line>Institute of Systems Biology, Universiti Kebangsaan Malaysia (UKM), 43600 UKM, Bangi, Selangor, Malaysia</addr-line></nlm:aff><country xml:lang="fr">Malaisie</country><wicri:regionArea>Institute of Systems Biology, Universiti Kebangsaan Malaysia (UKM), 43600 UKM, Bangi, Selangor</wicri:regionArea><wicri:noRegion>Selangor</wicri:noRegion></affiliation></author></analytic><series><title level="j">PLoS ONE</title><idno type="eISSN">1932-6203</idno><imprint><date when="2015">2015</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>Juvenile hormones have attracted attention as safe and selective targets for the design and development of environmentally friendly and biorational insecticides. In the juvenile hormone III biosynthetic pathway, the enzyme farnesol dehydrogenase catalyzes the oxidation of farnesol to farnesal. In this study, farnesol dehydrogenase was extracted from <italic>Polygonum minus</italic> leaves and purified 204-fold to apparent homogeneity by ion-exchange chromatography using DEAE-Toyopearl, SP-Toyopearl, and Super-Q Toyopearl, followed by three successive purifications by gel filtration chromatography on a TSK-gel GS3000SW. The enzyme is a heterodimer comprised of subunits with molecular masses of 65 kDa and 70 kDa. The optimum temperature and pH were 35°C and pH 9.5, respectively. Activity was inhibited by sulfhydryl reagents, metal-chelating agents and heavy metal ions. The enzyme utilized both NAD<sup>+</sup> and NADP<sup>+</sup> as coenzymes with <italic>K</italic><sub>m</sub> values of 0.74 mM and 40 mM, respectively. <italic>Trans</italic>, <italic>trans</italic>-farnesol was the preferred substrate for the <italic>P</italic>. <italic>minus</italic> farnesol dehydrogenase. Geometrical isomers of <italic>trans</italic>, <italic>trans</italic>-farnesol, <italic>cis</italic>, <italic>trans</italic>-farnesol and <italic>cis</italic>, <italic>cis</italic>-farnesol were also oxidized by the enzyme with lower activity. The <italic>K</italic><sub>m</sub> values for t<italic>rans</italic>, <italic>trans</italic>-farnesol, <italic>cis</italic>, <italic>trans</italic>-farnesol and <italic>cis</italic>, <italic>cis</italic>-farnesol appeared to be 0.17 mM, 0.33 mM and 0.42 mM, respectively. The amino acid sequences of 4 tryptic peptides of the enzyme were analyzed by MALDI-TOF/TOF-MS spectrometry, and showed no significant similarity to those of previously reported farnesol dehydrogenases. These results suggest that the purified enzyme is a novel NAD(P)<sup>+</sup>-dependent farnesol dehydrogenase. The purification and characterization established in the current study will serve as a basis to provide new information for recombinant production of the enzyme. Therefore, recombinant farnesol dehydrogenase may provide a useful molecular tool in manipulating juvenile hormone biosynthesis to generate transgenic plants for pest control.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Kamita, Sg" uniqKey="Kamita S">SG Kamita</name></author><author><name sortKey="Hammock, Bd" uniqKey="Hammock B">BD Hammock</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Lopez, O" uniqKey="Lopez O">O Lopez</name></author><author><name sortKey="Jose, Gf" uniqKey="Jose G">GF Jose</name></author><author><name sortKey="Maria, Vg" uniqKey="Maria V">VG Maria</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Hoffmann, Kh" uniqKey="Hoffmann K">KH Hoffmann</name></author><author><name sortKey="Lorenz, Mw" uniqKey="Lorenz M">MW Lorenz</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Furuta, K" uniqKey="Furuta K">K 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<front><journal-meta><journal-id journal-id-type="nlm-ta">PLoS One</journal-id><journal-id journal-id-type="iso-abbrev">PLoS ONE</journal-id><journal-id journal-id-type="publisher-id">plos</journal-id><journal-id journal-id-type="pmc">plosone</journal-id><journal-title-group><journal-title>PLoS ONE</journal-title></journal-title-group><issn pub-type="epub">1932-6203</issn><publisher><publisher-name>Public Library of Science</publisher-name><publisher-loc>San Francisco, CA USA</publisher-loc></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">26600471</article-id><article-id pub-id-type="pmc">4657912</article-id><article-id pub-id-type="doi">10.1371/journal.pone.0143310</article-id><article-id pub-id-type="publisher-id">PONE-D-15-31717</article-id><article-categories><subj-group subj-group-type="heading"><subject>Research Article</subject></subj-group></article-categories><title-group><article-title>Purification and Characterization of a Novel NAD(P)<sup>+</sup>-Farnesol Dehydrogenase from <italic>Polygonum minus</italic> Leaves</article-title><alt-title alt-title-type="running-head">Purification & Characterization of a Novel Farnesol-DH from <italic>P</italic>. <italic>minus</italic></alt-title></title-group><contrib-group><contrib contrib-type="author" equal-contrib="yes"><name><surname>Ahmad-Sohdi</surname><given-names>Nor-Ain-Shahajar</given-names></name><xref ref-type="aff" rid="aff001"><sup>1</sup></xref></contrib><contrib contrib-type="author"><name><surname>Seman-Kamarulzaman</surname><given-names>Ahmad-Faris</given-names></name><xref ref-type="aff" rid="aff001"><sup>1</sup></xref></contrib><contrib contrib-type="author"><name><surname>Mohamed-Hussein</surname><given-names>Zeti-Azura</given-names></name><xref ref-type="aff" rid="aff001"><sup>1</sup></xref><xref ref-type="aff" rid="aff002"><sup>2</sup></xref></contrib><contrib contrib-type="author" equal-contrib="yes"><name><surname>Hassan</surname><given-names>Maizom</given-names></name><xref ref-type="aff" rid="aff001"><sup>1</sup></xref><xref rid="cor001" ref-type="corresp">*</xref></contrib></contrib-group><aff id="aff001"><label>1</label><addr-line>Institute of Systems Biology, Universiti Kebangsaan Malaysia (UKM), 43600 UKM, Bangi, Selangor, Malaysia</addr-line></aff><aff id="aff002"><label>2</label><addr-line>School of Biosciences and Biotechnology, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, 43600 UKM, Bangi, Selangor, Malaysia</addr-line></aff><contrib-group><contrib contrib-type="editor"><name><surname>Motta</surname><given-names>Andrea</given-names></name><role>Editor</role><xref ref-type="aff" rid="edit1"></xref></contrib></contrib-group><aff id="edit1"><addr-line>National Research Council of Italy, ITALY</addr-line></aff><author-notes><fn fn-type="conflict" id="coi001"><p><bold>Competing Interests: </bold>The authors have declared that no competing interests exist.</p></fn><fn fn-type="con" id="contrib001"><p>Conceived and designed the experiments: NASAS MH ZAMH. Performed the experiments: NASAS AFSK. Analyzed the data: NASAS MH ZAMH AFSK. Contributed reagents/materials/analysis tools: MH ZAMH. Wrote the paper: NASAS MH ZAMH AFSK.</p></fn><corresp id="cor001">* E-mail: <email>maizom@ukm.edu.my</email></corresp></author-notes><pub-date pub-type="epub"><day>23</day><month>11</month><year>2015</year></pub-date><pub-date pub-type="collection"><year>2015</year></pub-date><volume>10</volume><issue>11</issue><elocation-id>e0143310</elocation-id><history><date date-type="received"><day>19</day><month>7</month><year>2015</year></date><date date-type="accepted"><day>3</day><month>11</month><year>2015</year></date></history><permissions><copyright-year>2015</copyright-year><copyright-holder>Ahmad-Sohdi et al</copyright-holder><license xlink:href="http://creativecommons.org/licenses/by/4.0/"><license-p>This is an open access article distributed under the terms of the <ext-link ext-link-type="uri" xlink:href="http://creativecommons.org/licenses/by/4.0/">Creative Commons Attribution License</ext-link>, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited</license-p></license></permissions><self-uri content-type="pdf" xlink:type="simple" xlink:href="pone.0143310.pdf"></self-uri><abstract><p>Juvenile hormones have attracted attention as safe and selective targets for the design and development of environmentally friendly and biorational insecticides. In the juvenile hormone III biosynthetic pathway, the enzyme farnesol dehydrogenase catalyzes the oxidation of farnesol to farnesal. In this study, farnesol dehydrogenase was extracted from <italic>Polygonum minus</italic> leaves and purified 204-fold to apparent homogeneity by ion-exchange chromatography using DEAE-Toyopearl, SP-Toyopearl, and Super-Q Toyopearl, followed by three successive purifications by gel filtration chromatography on a TSK-gel GS3000SW. The enzyme is a heterodimer comprised of subunits with molecular masses of 65 kDa and 70 kDa. The optimum temperature and pH were 35°C and pH 9.5, respectively. Activity was inhibited by sulfhydryl reagents, metal-chelating agents and heavy metal ions. The enzyme utilized both NAD<sup>+</sup> and NADP<sup>+</sup> as coenzymes with <italic>K</italic><sub>m</sub> values of 0.74 mM and 40 mM, respectively. <italic>Trans</italic>, <italic>trans</italic>-farnesol was the preferred substrate for the <italic>P</italic>. <italic>minus</italic> farnesol dehydrogenase. Geometrical isomers of <italic>trans</italic>, <italic>trans</italic>-farnesol, <italic>cis</italic>, <italic>trans</italic>-farnesol and <italic>cis</italic>, <italic>cis</italic>-farnesol were also oxidized by the enzyme with lower activity. The <italic>K</italic><sub>m</sub> values for t<italic>rans</italic>, <italic>trans</italic>-farnesol, <italic>cis</italic>, <italic>trans</italic>-farnesol and <italic>cis</italic>, <italic>cis</italic>-farnesol appeared to be 0.17 mM, 0.33 mM and 0.42 mM, respectively. The amino acid sequences of 4 tryptic peptides of the enzyme were analyzed by MALDI-TOF/TOF-MS spectrometry, and showed no significant similarity to those of previously reported farnesol dehydrogenases. These results suggest that the purified enzyme is a novel NAD(P)<sup>+</sup>-dependent farnesol dehydrogenase. The purification and characterization established in the current study will serve as a basis to provide new information for recombinant production of the enzyme. Therefore, recombinant farnesol dehydrogenase may provide a useful molecular tool in manipulating juvenile hormone biosynthesis to generate transgenic plants for pest control.</p></abstract><funding-group><funding-statement>This work was funded by the Ministry of Science, Technology & Innovation (Grant number: 02-01-02-SF0852, URL: <ext-link ext-link-type="uri" xlink:href="http://www.mosti.gov.my/">http://www.mosti.gov.my/</ext-link>), and International Foundation for Science (Grant number: RB-IFS-001-2012, URL: <ext-link ext-link-type="uri" xlink:href="http://www.ifs.se/">http://www.ifs.se/</ext-link>). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.</funding-statement></funding-group><counts><fig-count count="4"></fig-count><table-count count="4"></table-count><page-count count="18"></page-count></counts><custom-meta-group><custom-meta id="data-availability"><meta-name>Data Availability</meta-name><meta-value>All relevant data are within the paper and its Supporting Information files.</meta-value></custom-meta></custom-meta-group></article-meta><notes><title>Data Availability</title><p>All relevant data are within the paper and its Supporting Information files.</p></notes></front></pmc><affiliations><list><country><li>Malaisie</li></country></list><tree><country name="Malaisie"><noRegion><name sortKey="Ahmad Sohdi, Nor Ain Shahajar" sort="Ahmad Sohdi, Nor Ain Shahajar" uniqKey="Ahmad Sohdi N" first="Nor-Ain-Shahajar" last="Ahmad-Sohdi">Nor-Ain-Shahajar Ahmad-Sohdi</name></noRegion><name sortKey="Hassan, Maizom" sort="Hassan, Maizom" uniqKey="Hassan M" first="Maizom" last="Hassan">Maizom Hassan</name><name sortKey="Mohamed Hussein, Zeti Azura" sort="Mohamed Hussein, Zeti Azura" uniqKey="Mohamed Hussein Z" first="Zeti-Azura" last="Mohamed-Hussein">Zeti-Azura Mohamed-Hussein</name><name sortKey="Mohamed Hussein, Zeti Azura" sort="Mohamed Hussein, Zeti Azura" uniqKey="Mohamed Hussein Z" first="Zeti-Azura" last="Mohamed-Hussein">Zeti-Azura Mohamed-Hussein</name><name sortKey="Seman Kamarulzaman, Ahmad Faris" sort="Seman Kamarulzaman, Ahmad Faris" uniqKey="Seman Kamarulzaman A" first="Ahmad-Faris" last="Seman-Kamarulzaman">Ahmad-Faris Seman-Kamarulzaman</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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