A Recoverable Enzymatic Microgel Based on Biomolecular Recognition
The enzyme β-galactosidase has been immobilized through incorporation into a selectively soluble microgel, prepared from DNA, biotinylated peptide nucleic acid (PNA), and the protein avidin. The enzyme was conjugated to avidin, allowing it to be integrated directly into the microgel network. Efficie...
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| Veröffentlicht in: | Journal of the American Chemical Society 2004-01, Vol.126 (3), p.726-727 |
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| container_end_page | 727 |
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| container_issue | 3 |
| container_start_page | 726 |
| container_title | Journal of the American Chemical Society |
| container_volume | 126 |
| creator | Cao, Rong Gu, Zhenyu Patterson, Gary D Armitage, Bruce A |
| description | The enzyme β-galactosidase has been immobilized through incorporation into a selectively soluble microgel, prepared from DNA, biotinylated peptide nucleic acid (PNA), and the protein avidin. The enzyme was conjugated to avidin, allowing it to be integrated directly into the microgel network. Efficient hydrolysis of a small-molecule substrate occurred at 37°, but cooling and centrifuging led to precipitation of the microgels and product separation. The microgels were then reconstituted by adding fresh buffer and shaking. The enzymatic activity was completely recovered through repeated cycles. This method should be generalizable to a wide variety of other enzymes and substrates. |
| doi_str_mv | 10.1021/ja039502d |
| format | Article |
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The enzyme was conjugated to avidin, allowing it to be integrated directly into the microgel network. Efficient hydrolysis of a small-molecule substrate occurred at 37°, but cooling and centrifuging led to precipitation of the microgels and product separation. The microgels were then reconstituted by adding fresh buffer and shaking. The enzymatic activity was completely recovered through repeated cycles. This method should be generalizable to a wide variety of other enzymes and substrates.</description><identifier>ISSN: 0002-7863</identifier><identifier>EISSN: 1520-5126</identifier><identifier>DOI: 10.1021/ja039502d</identifier><identifier>PMID: 14733541</identifier><identifier>CODEN: JACSAT</identifier><language>eng</language><publisher>Washington, DC: American Chemical Society</publisher><subject>Avidin - chemistry ; beta-Galactosidase - chemistry ; beta-Galactosidase - metabolism ; Biological and medical sciences ; Biotechnology ; Biotin - chemistry ; DNA - chemistry ; Enzymes, Immobilized - chemistry ; Enzymes, Immobilized - metabolism ; Fundamental and applied biological sciences. Psychology ; Gels ; Immobilization of enzymes and other molecules ; Immobilization techniques ; Kinetics ; Methods. Procedures. Technologies ; Nitrophenylgalactosides - chemistry ; Nitrophenylgalactosides - metabolism ; Nucleic Acid Conformation</subject><ispartof>Journal of the American Chemical Society, 2004-01, Vol.126 (3), p.726-727</ispartof><rights>Copyright © 2004 American Chemical Society</rights><rights>2004 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-a379t-5c93e490810853e571cdcf2ff8e5c6da7506371c08d7b16450092f120a35406b3</citedby><cites>FETCH-LOGICAL-a379t-5c93e490810853e571cdcf2ff8e5c6da7506371c08d7b16450092f120a35406b3</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/ja039502d$$EPDF$$P50$$Gacs$$H</linktopdf><linktohtml>$$Uhttps://pubs.acs.org/doi/10.1021/ja039502d$$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=15422505$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/14733541$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Cao, Rong</creatorcontrib><creatorcontrib>Gu, Zhenyu</creatorcontrib><creatorcontrib>Patterson, Gary D</creatorcontrib><creatorcontrib>Armitage, Bruce A</creatorcontrib><title>A Recoverable Enzymatic Microgel Based on Biomolecular Recognition</title><title>Journal of the American Chemical Society</title><addtitle>J. Am. Chem. Soc</addtitle><description>The enzyme β-galactosidase has been immobilized through incorporation into a selectively soluble microgel, prepared from DNA, biotinylated peptide nucleic acid (PNA), and the protein avidin. The enzyme was conjugated to avidin, allowing it to be integrated directly into the microgel network. Efficient hydrolysis of a small-molecule substrate occurred at 37°, but cooling and centrifuging led to precipitation of the microgels and product separation. The microgels were then reconstituted by adding fresh buffer and shaking. The enzymatic activity was completely recovered through repeated cycles. This method should be generalizable to a wide variety of other enzymes and substrates.</description><subject>Avidin - chemistry</subject><subject>beta-Galactosidase - chemistry</subject><subject>beta-Galactosidase - metabolism</subject><subject>Biological and medical sciences</subject><subject>Biotechnology</subject><subject>Biotin - chemistry</subject><subject>DNA - chemistry</subject><subject>Enzymes, Immobilized - chemistry</subject><subject>Enzymes, Immobilized - metabolism</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gels</subject><subject>Immobilization of enzymes and other molecules</subject><subject>Immobilization techniques</subject><subject>Kinetics</subject><subject>Methods. Procedures. Technologies</subject><subject>Nitrophenylgalactosides - chemistry</subject><subject>Nitrophenylgalactosides - metabolism</subject><subject>Nucleic Acid Conformation</subject><issn>0002-7863</issn><issn>1520-5126</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2004</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNptkM1OwzAQhC0EglI48AIoF5A4BNZ2HCfHtioUqRWIH3G0XMepXJIY7AQBT4-hVXvhtNrdb0ezg9AJhksMBF8tJdCcASl2UA8zAjHDJN1FPQAgMc9SeoAOvV-GNiEZ3kcHOOGUsgT30HAQPWhlP7ST80pH4-b7q5atUdHMKGcXuoqG0usisk00NLa2lVZdJd3f0aIxrbHNEdorZeX18br20fP1-Gk0iad3N7ejwTSWlOdtzFROdZJDhiFjVDOOVaFKUpaZZiotJGeQ0jCErOBznCYMICclJiCDU0jntI_OV7pvzr532reiNl7pqpKNtp0XGWBI8vBYH12swPCB906X4s2ZWrovgUH8BiY2gQX2dC3azWtdbMl1QgE4WwPSK1mVTjbK-C3HEkIYsMDFK874Vn9u9tK9ipRTzsTT_aOgN6PJC3_AYrbVlcqLpe1cE7L7x-APr8aLgw</recordid><startdate>20040128</startdate><enddate>20040128</enddate><creator>Cao, Rong</creator><creator>Gu, Zhenyu</creator><creator>Patterson, Gary D</creator><creator>Armitage, Bruce A</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>7X8</scope></search><sort><creationdate>20040128</creationdate><title>A Recoverable Enzymatic Microgel Based on Biomolecular Recognition</title><author>Cao, Rong ; Gu, Zhenyu ; Patterson, Gary D ; Armitage, Bruce A</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-a379t-5c93e490810853e571cdcf2ff8e5c6da7506371c08d7b16450092f120a35406b3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2004</creationdate><topic>Avidin - chemistry</topic><topic>beta-Galactosidase - chemistry</topic><topic>beta-Galactosidase - metabolism</topic><topic>Biological and medical sciences</topic><topic>Biotechnology</topic><topic>Biotin - chemistry</topic><topic>DNA - chemistry</topic><topic>Enzymes, Immobilized - chemistry</topic><topic>Enzymes, Immobilized - metabolism</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Gels</topic><topic>Immobilization of enzymes and other molecules</topic><topic>Immobilization techniques</topic><topic>Kinetics</topic><topic>Methods. Procedures. Technologies</topic><topic>Nitrophenylgalactosides - chemistry</topic><topic>Nitrophenylgalactosides - metabolism</topic><topic>Nucleic Acid Conformation</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Cao, Rong</creatorcontrib><creatorcontrib>Gu, Zhenyu</creatorcontrib><creatorcontrib>Patterson, Gary D</creatorcontrib><creatorcontrib>Armitage, Bruce A</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>MEDLINE - Academic</collection><jtitle>Journal of the American Chemical Society</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Cao, Rong</au><au>Gu, Zhenyu</au><au>Patterson, Gary D</au><au>Armitage, Bruce A</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A Recoverable Enzymatic Microgel Based on Biomolecular Recognition</atitle><jtitle>Journal of the American Chemical Society</jtitle><addtitle>J. Am. Chem. Soc</addtitle><date>2004-01-28</date><risdate>2004</risdate><volume>126</volume><issue>3</issue><spage>726</spage><epage>727</epage><pages>726-727</pages><issn>0002-7863</issn><eissn>1520-5126</eissn><coden>JACSAT</coden><abstract>The enzyme β-galactosidase has been immobilized through incorporation into a selectively soluble microgel, prepared from DNA, biotinylated peptide nucleic acid (PNA), and the protein avidin. The enzyme was conjugated to avidin, allowing it to be integrated directly into the microgel network. Efficient hydrolysis of a small-molecule substrate occurred at 37°, but cooling and centrifuging led to precipitation of the microgels and product separation. The microgels were then reconstituted by adding fresh buffer and shaking. The enzymatic activity was completely recovered through repeated cycles. This method should be generalizable to a wide variety of other enzymes and substrates.</abstract><cop>Washington, DC</cop><pub>American Chemical Society</pub><pmid>14733541</pmid><doi>10.1021/ja039502d</doi><tpages>2</tpages></addata></record> |
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| ispartof | Journal of the American Chemical Society, 2004-01, Vol.126 (3), p.726-727 |
| issn | 0002-7863 1520-5126 |
| language | eng |
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| source | MEDLINE; American Chemical Society Journals |
| subjects | Avidin - chemistry beta-Galactosidase - chemistry beta-Galactosidase - metabolism Biological and medical sciences Biotechnology Biotin - chemistry DNA - chemistry Enzymes, Immobilized - chemistry Enzymes, Immobilized - metabolism Fundamental and applied biological sciences. Psychology Gels Immobilization of enzymes and other molecules Immobilization techniques Kinetics Methods. Procedures. Technologies Nitrophenylgalactosides - chemistry Nitrophenylgalactosides - metabolism Nucleic Acid Conformation |
| title | A Recoverable Enzymatic Microgel Based on Biomolecular Recognition |
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