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<!DOCTYPE html> <html lang="en" dir="ltr" xmlns="http://www.w3.org/1999/xhtml" xmlns:mml="http://www.w3.org/1998/Math/MathML"> <head prefix="og: http://ogp.me/ns# article: http://ogp.me/ns/article# book: http://ogp.me/ns/book#" >  <link rel="dns-prefetch" href="//d33xdlntwy0kbs.cloudfront.net" /> <link rel="dns-prefetch" href="//www.google.com" /> <link rel="dns-prefetch" href="//scholar.google.com" /> <link rel="dns-prefetch" href="//www.googletagmanager.com" /> <meta http-equiv="Content-Type" content="text/html; charset=utf-8" /> <link rel="shortcut icon" href="https://www.biorxiv.org/sites/default/files/images/favicon.ico" type="image/vnd.microsoft.icon" /> <meta name="viewport" content="width=device-width, initial-scale=1" /> <link rel="alternate" type="application/pdf" title="Full Text (PDF)" href="/content/10.1101/2020.07.24.220400v2.full.pdf" /> <link rel="alternate" type="text/plain" title="Full Text (Plain)" href="/content/10.1101/2020.07.24.220400v2.full.txt" /> <meta name="type" content="article" /> <meta name="category" content="article" /> <meta name="HW.identifier" content="/biorxiv/early/2020/12/03/2020.07.24.220400.atom" /> <meta name="HW.pisa" content="biorxiv;2020.07.24.220400v2" /> <meta name="DC.Format" content="text/html" /> <meta name="DC.Language" content="en" /> <meta name="DC.Title" content="Separation and quantification of 2-Keto-3-deoxy-gluconate (KDG) a major metabolite in pectin and alginate degradation pathways" /> <meta name="DC.Identifier" content="10.1101/2020.07.24.220400" /> <meta name="DC.Date" content="2020-12-03" /> <meta name="DC.Publisher" content="Cold Spring Harbor Laboratory" /> <meta name="DC.Rights" content="© 2020, Posted by Cold Spring Harbor Laboratory. This pre-print is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), CC BY-NC 4.0, as described at http://creativecommons.org/licenses/by-nc/4.0/" /> <meta name="DC.AccessRights" content="restricted" /> <meta name="DC.Description" content="A rapid and sensitive High Performance Liquid Chromatography (HPLC) method with photometric and fluorescence detection is developed for routine analysis of 2-Keto-3-deoxy-gluconate (KDG), a catabolite product of pectin and alginate. These polysaccharides are primary-based compounds for biofuel production and for generation of high-value-added products. HPLC is performed, after derivatization of the 2-oxo-acid groups of the metabolite with o-phenylenediamine (oPD), using a linear gradient of trifluoroacetic acid and acetonitrile. Quantification is accomplished with an internal standard method. The gradient is optimized to distinguish KDG from its close structural analogues such as 5-keto-4-deoxyuronate (DKI) and 2,5-diketo-3-deoxygluconate (DKII). The proposed method is simple, highly sensitive and accurate for time course analysis of pectin or alginate degradation. Highlights A fluorescent based-HPLC method report the quantification of KDG, a metabolite originating from alginate and from pectin degradation pathways, using derivatization with o-phenylenediamine (oPD) ### Competing Interest Statement The authors have declared no competing interest." /> <meta name="DC.Contributor" content="Shiny Martis B" /> <meta name="DC.Contributor" content="Michel Droux" /> <meta name="DC.Contributor" content="Florelle Deboudard" /> <meta name="DC.Contributor" content="William Nasser" /> <meta name="DC.Contributor" content="Sam Meyer" /> <meta name="DC.Contributor" content="Sylvie Reverchon" /> <meta name="article:published_time" content="2020-12-03" /> <meta name="article:section" content="New Results" /> <meta name="citation_title" content="Separation and quantification of 2-Keto-3-deoxy-gluconate (KDG) a major metabolite in pectin and alginate degradation pathways" /> <meta name="citation_abstract" lang="en" content="<h3>Abstract</h3> <p>A rapid and sensitive High Performance Liquid Chromatography (HPLC) method with photometric and fluorescence detection is developed for routine analysis of 2-Keto-3-deoxy-gluconate (KDG), a catabolite product of pectin and alginate. These polysaccharides are primary-based compounds for biofuel production and for generation of high-value-added products. HPLC is performed, after derivatization of the 2-oxo-acid groups of the metabolite with o-phenylenediamine (oPD), using a linear gradient of trifluoroacetic acid and acetonitrile. Quantification is accomplished with an internal standard method. The gradient is optimized to distinguish KDG from its close structural analogues such as 5-keto-4-deoxyuronate (DKI) and 2,5-diketo-3-deoxygluconate (DKII). The proposed method is simple, highly sensitive and accurate for time course analysis of pectin or alginate degradation.</p><h3>Highlights</h3> <p>A fluorescent based-HPLC method report the quantification of KDG, a metabolite originating from alginate and from pectin degradation pathways, using derivatization with o-phenylenediamine (oPD)</p>" /> <meta name="citation_journal_title" content="bioRxiv" /> <meta name="citation_publisher" content="Cold Spring Harbor Laboratory" /> <meta name="citation_publication_date" content="2020/01/01" /> <meta name="citation_mjid" content="biorxiv;2020.07.24.220400v2" /> <meta name="citation_id" content="2020.07.24.220400v2" /> <meta name="citation_public_url" content="https://www.biorxiv.org/content/10.1101/2020.07.24.220400v2" /> <meta name="citation_abstract_html_url" content="https://www.biorxiv.org/content/10.1101/2020.07.24.220400v2.abstract" /> <meta name="citation_full_html_url" content="https://www.biorxiv.org/content/10.1101/2020.07.24.220400v2.full" /> <meta name="citation_pdf_url" content="https://www.biorxiv.org/content/biorxiv/early/2020/12/03/2020.07.24.220400.full.pdf" /> <meta name="citation_doi" content="10.1101/2020.07.24.220400" /> <meta name="citation_num_pages" content="31" /> <meta name="citation_article_type" content="Article" /> <meta name="citation_section" content="New Results" /> <meta name="citation_firstpage" content="2020.07.24.220400" /> <meta name="citation_author" content="Shiny Martis B" /> <meta name="citation_author_institution" content="Univ Lyon, Université Claude Bernard Lyon 1" /> <meta name="citation_author" content="Michel Droux" /> <meta name="citation_author_institution" content="Univ Lyon, Université Claude Bernard Lyon 1" /> <meta name="citation_author_email" content="michel.droux@univ-lyon1.fr" /> <meta name="citation_author_orcid" content="http://orcid.org/0000-0001-5635-2046" /> <meta name="citation_author" content="Florelle Deboudard" /> <meta name="citation_author_institution" content="Univ Lyon, Université Claude Bernard Lyon 1" /> <meta name="citation_author" content="William Nasser" /> <meta name="citation_author_institution" content="Univ Lyon, Université Claude Bernard Lyon 1" /> <meta name="citation_author_orcid" content="http://orcid.org/0000-0001-9848-3597" /> <meta name="citation_author" content="Sam Meyer" /> <meta name="citation_author_institution" content="Univ Lyon, Université Claude Bernard Lyon 1" /> <meta name="citation_author_orcid" content="http://orcid.org/0000-0001-7206-6226" /> <meta name="citation_author" content="Sylvie Reverchon" /> <meta name="citation_author_institution" content="Univ Lyon, Université Claude Bernard Lyon 1" /> <meta name="citation_author_orcid" content="http://orcid.org/0000-0002-0478-3474" /> <meta name="citation_reference" content="M.E. 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Valle, Metabolically engineered bacterial strains having enhanced 2-keto-D-gluconate accumulation, US20050106693A1 (2005)." /> <meta name="twitter:title" content="Separation and quantification of 2-Keto-3-deoxy-gluconate (KDG) a major metabolite in pectin and alginate degradation pathways" /> <meta name="twitter:site" content="@biorxivpreprint" /> <meta name="twitter:card" content="summary" /> <meta name="twitter:image" content="https://www.biorxiv.org/sites/default/files/images/biorxiv_logo_homepage7-5-small.png" /> <meta name="twitter:description" content="A rapid and sensitive High Performance Liquid Chromatography (HPLC) method with photometric and fluorescence detection is developed for routine analysis of 2-Keto-3-deoxy-gluconate (KDG), a catabolite product of pectin and alginate. These polysaccharides are primary-based compounds for biofuel production and for generation of high-value-added products. HPLC is performed, after derivatization of the 2-oxo-acid groups of the metabolite with o-phenylenediamine (oPD), using a linear gradient of trifluoroacetic acid and acetonitrile. Quantification is accomplished with an internal standard method. The gradient is optimized to distinguish KDG from its close structural analogues such as 5-keto-4-deoxyuronate (DKI) and 2,5-diketo-3-deoxygluconate (DKII). The proposed method is simple, highly sensitive and accurate for time course analysis of pectin or alginate degradation. Highlights A fluorescent based-HPLC method report the quantification of KDG, a metabolite originating from alginate and from pectin degradation pathways, using derivatization with o-phenylenediamine (oPD) ### Competing Interest Statement The authors have declared no competing interest." /> <meta name="og-title" property="og:title" content="Separation and quantification of 2-Keto-3-deoxy-gluconate (KDG) a major metabolite in pectin and alginate degradation pathways" /> <meta name="og-url" property="og:url" content="https://www.biorxiv.org/content/10.1101/2020.07.24.220400v2" /> <meta name="og-site-name" property="og:site_name" content="bioRxiv" /> <meta name="og-description" property="og:description" content="A rapid and sensitive High Performance Liquid Chromatography (HPLC) method with photometric and fluorescence detection is developed for routine analysis of 2-Keto-3-deoxy-gluconate (KDG), a catabolite product of pectin and alginate. These polysaccharides are primary-based compounds for biofuel production and for generation of high-value-added products. HPLC is performed, after derivatization of the 2-oxo-acid groups of the metabolite with o-phenylenediamine (oPD), using a linear gradient of trifluoroacetic acid and acetonitrile. Quantification is accomplished with an internal standard method. The gradient is optimized to distinguish KDG from its close structural analogues such as 5-keto-4-deoxyuronate (DKI) and 2,5-diketo-3-deoxygluconate (DKII). The proposed method is simple, highly sensitive and accurate for time course analysis of pectin or alginate degradation. Highlights A fluorescent based-HPLC method report the quantification of KDG, a metabolite originating from alginate and from pectin degradation pathways, using derivatization with o-phenylenediamine (oPD) ### Competing Interest Statement The authors have declared no competing interest." /> <meta name="og-type" property="og:type" content="article" /> <meta name="og-image" property="og:image" content="https://www.biorxiv.org/sites/default/files/images/biorxiv_logo_homepage7-5-small.png" /> <meta name="citation_date" content="2020-12-03" /> <link rel="alternate" type="application/vnd.ms-powerpoint" title="Powerpoint" href="/content/10.1101/2020.07.24.220400v2.ppt" /> <meta name="description" content="bioRxiv - the preprint server for biology, operated by Cold Spring Harbor Laboratory, a research and educational institution" /> <meta name="generator" content="Drupal 7 (http://drupal.org)" /> <link rel="canonical" href="https://www.biorxiv.org/content/10.1101/2020.07.24.220400v2" /> <link 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class="highwire-article-citation highwire-citation-type-highwire-article node1666400" data-node-nid="1666400" id="node-1666400--2717979114" data-pisa="biorxiv;2020.07.24.220400v2" data-pisa-master="biorxiv;2020.07.24.220400" data-apath="/biorxiv/early/2020/12/03/2020.07.24.220400.atom" data-hw-author-tooltip-instance="highwire_author_tooltip"><div class="highwire-cite highwire-cite-highwire-article highwire-citation-biorxiv-article-top clearfix has-author-tooltip" > <span class="biorxiv-article-type"> New Results </span> <h1 class="highwire-cite-title" id="page-title">Separation and quantification of 2-Keto-3-deoxy-gluconate (KDG) a major metabolite in pectin and alginate degradation pathways</h1> <div class="highwire-cite-authors" ><span class="highwire-citation-authors"><span class="highwire-citation-author first" data-delta="0"><span class="nlm-given-names">Shiny</span> <span class="nlm-surname">Martis B</span></span>, <span class="highwire-citation-author hw-author-orcid-logo-wrapper" data-delta="1"><a href="http://orcid.org/0000-0001-5635-2046" target="_blank" class="hw-author-orcid-logo link-icon-only link-icon"><span class="hw-icon-orcid hw-icon-color-orcid"></span> <span class="title element-invisible">View ORCID Profile</span></a><span class="nlm-given-names">Michel</span> <span class="nlm-surname">Droux</span></span>, <span class="highwire-citation-author" data-delta="2"><span class="nlm-given-names">Florelle</span> <span class="nlm-surname">Deboudard</span></span>, <span class="highwire-citation-author hw-author-orcid-logo-wrapper" data-delta="3"><a href="http://orcid.org/0000-0001-9848-3597" target="_blank" class="hw-author-orcid-logo link-icon-only link-icon"><span class="hw-icon-orcid hw-icon-color-orcid"></span> <span class="title element-invisible">View ORCID Profile</span></a><span class="nlm-given-names">William</span> <span class="nlm-surname">Nasser</span></span>, <span class="highwire-citation-author hw-author-orcid-logo-wrapper" data-delta="4"><a href="http://orcid.org/0000-0001-7206-6226" target="_blank" class="hw-author-orcid-logo link-icon-only link-icon"><span class="hw-icon-orcid hw-icon-color-orcid"></span> <span class="title element-invisible">View ORCID Profile</span></a><span class="nlm-given-names">Sam</span> <span class="nlm-surname">Meyer</span></span>, <span class="highwire-citation-author hw-author-orcid-logo-wrapper" data-delta="5"><a href="http://orcid.org/0000-0002-0478-3474" target="_blank" class="hw-author-orcid-logo link-icon-only link-icon"><span class="hw-icon-orcid hw-icon-color-orcid"></span> <span class="title element-invisible">View ORCID Profile</span></a><span class="nlm-given-names">Sylvie</span> <span class="nlm-surname">Reverchon</span></span></span></div> <div class="highwire-cite-metadata" ><span class="highwire-cite-metadata-doi highwire-cite-metadata"><span class="label">doi:</span> https://doi.org/10.1101/2020.07.24.220400 </span></div> </div> <div id="hw-article-author-popups-node-1666400--2717979114" style="display: none;"><div class="author-tooltip-0"><div class="author-tooltip-name">Shiny Martis B </div><div class="author-tooltip-affiliation"><span class="author-tooltip-text"><div class='author-affiliation'><span class='nlm-sup'>a</span><span class='nlm-institution'>Univ Lyon, Université Claude Bernard Lyon 1</span>, INSA-Lyon, CNRS, UMR5240 MAP, F-69621, <span class='nlm-country'>France</span></div></span></div><ul class="author-tooltip-find-more"><li class="author-tooltip-gs-link first"><a href="/lookup/google-scholar?link_type=googlescholar&gs_type=author&author%5B0%5D=Shiny%2BMartis%2BB%2B" target="_blank" class="" data-icon-position="" data-hide-link-title="0">Find this author on Google Scholar</a></li><li class="author-tooltip-pubmed-link"><a href="/lookup/external-ref?access_num=Martis%20B%20S&link_type=AUTHORSEARCH" target="_blank" class="" data-icon-position="" data-hide-link-title="0">Find this author on 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class='nlm-sup'>a</span><span class='nlm-institution'>Univ Lyon, Université Claude Bernard Lyon 1</span>, INSA-Lyon, CNRS, UMR5240 MAP, F-69621, <span class='nlm-country'>France</span></div></span></div><ul class="author-tooltip-find-more"><li class="author-tooltip-gs-link first"><a href="/lookup/google-scholar?link_type=googlescholar&gs_type=author&author%5B0%5D=Florelle%2BDeboudard%2B" target="_blank" class="" data-icon-position="" data-hide-link-title="0">Find this author on Google Scholar</a></li><li class="author-tooltip-pubmed-link"><a href="/lookup/external-ref?access_num=Deboudard%20F&link_type=AUTHORSEARCH" target="_blank" class="" data-icon-position="" data-hide-link-title="0">Find this author on PubMed</a></li><li class="author-site-search-link last"><a href="/search/author1%3AFlorelle%2BDeboudard%2B" rel="nofollow" class="" data-icon-position="" data-hide-link-title="0">Search for this author on this site</a></li></ul></div><div class="author-tooltip-3"><div class="author-tooltip-name">William Nasser </div><div class="author-tooltip-affiliation"><span class="author-tooltip-text"><div class='author-affiliation'><span class='nlm-sup'>a</span><span class='nlm-institution'>Univ Lyon, Université Claude Bernard Lyon 1</span>, INSA-Lyon, CNRS, UMR5240 MAP, F-69621, <span class='nlm-country'>France</span></div></span></div><ul class="author-tooltip-find-more"><li class="author-tooltip-gs-link first"><a href="/lookup/google-scholar?link_type=googlescholar&gs_type=author&author%5B0%5D=William%2BNasser%2B" target="_blank" class="" data-icon-position="" data-hide-link-title="0">Find this author on Google Scholar</a></li><li class="author-tooltip-pubmed-link"><a href="/lookup/external-ref?access_num=Nasser%20W&link_type=AUTHORSEARCH" target="_blank" class="" data-icon-position="" data-hide-link-title="0">Find this author on PubMed</a></li><li class="author-site-search-link"><a href="/search/author1%3AWilliam%2BNasser%2B" rel="nofollow" class="" 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These polysaccharides are primary-based compounds for biofuel production and for generation of high-value-added products. HPLC is performed, after derivatization of the 2-oxo-acid groups of the metabolite with o-phenylenediamine (oPD), using a linear gradient of trifluoroacetic acid and acetonitrile. Quantification is accomplished with an internal standard method. The gradient is optimized to distinguish KDG from its close structural analogues such as 5-keto-4-deoxyuronate (DKI) and 2,5-diketo-3-deoxygluconate (DKII). The proposed method is simple, highly sensitive and accurate for time course analysis of pectin or alginate degradation.</p><div id="sec-1" class="subsection"><p id="p-3"><strong>Highlights</strong> A fluorescent based-HPLC method report the quantification of KDG, a metabolite originating from alginate and from pectin degradation pathways, using derivatization with o-phenylenediamine (oPD)</p></div></div><h3>Competing Interest Statement</h3><p id="p-4">The authors have declared no competing interest.</p><span class="highwire-journal-article-marker-end"></span></div><span class="related-urls"></span></div></div> </div> </div> <div class="panel-separator"></div><div class="panel-pane pane-biorxiv-copyright" > <div class="pane-content"> <div class="field field-name-field-highwire-copyright field-type-text field-label-inline clearfix"><div class="field-label">Copyright </div><div class="field-items"><div class="field-item even">The copyright holder for this preprint is the author/funder, 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data-delta="1"><span class="nlm-given-names">Michel</span> <span class="nlm-surname">Droux</span></span>, <span class="highwire-citation-author" data-delta="2"><span class="nlm-given-names">Florelle</span> <span class="nlm-surname">Deboudard</span></span>, <span class="highwire-citation-author" data-delta="3"><span class="nlm-given-names">William</span> <span class="nlm-surname">Nasser</span></span>, <span class="highwire-citation-author" data-delta="4"><span class="nlm-given-names">Sam</span> <span class="nlm-surname">Meyer</span></span>, <span class="highwire-citation-author" data-delta="5"><span class="nlm-given-names">Sylvie</span> <span class="nlm-surname">Reverchon</span></span></span></div> <div class="highwire-cite-metadata" ><span class="highwire-cite-metadata-journal highwire-cite-metadata">bioRxiv </span><span class="highwire-cite-metadata-pages highwire-cite-metadata">2020.07.24.220400; </span><span class="highwire-cite-metadata-doi highwire-cite-metadata"><span class="doi_label">doi:</span> https://doi.org/10.1101/2020.07.24.220400 </span></div> </div> </div> </div> <div class="highwire-citation-formats"> <h2>Citation Manager Formats</h2> <div class="highwire-citation-formats-links"> <span class="Z3988" title="ctx_ver=Z39.88-2004&rft_val_fmt=info%3Aofi%2Ffmt%3Akev%3Amtx%3Ajournal&rft.spage&rft.epage&rft.atitle=Separation%20and%20quantification%20of%202-Keto-3-deoxy-gluconate%20%28KDG%29%20a%20major%20metabolite%20in%20pectin%20and%20alginate%20degradation%20pathways&rft.volume&rft.issue&rft.date=2020-01-01%2000%3A00%3A00&rft.stitle&rft.jtitle=bioRxiv&rft.au=Martis+B%2C+Shiny&rft.au=Droux%2C+Michel&rft.au=Deboudard%2C+Florelle&rft.au=Nasser%2C+William&rft.au=Meyer%2C+Sam&rft.au=Reverchon%2C+Sylvie"></span><ul class="hw-citation-links inline button button-alt button-grid clearfix"><li class="bibtext first"><a href="/highwire/citation/1666400/bibtext" class="hw-download-citation-link" data-icon-position="" 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