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Organosoluble enzyme conjugates with poly(2-oxazoline)s via pyromellitic acid dianhydride
[Display omitted] ► POXylation is successful via pyromellitic acid dianhydride. ► Hen-egg white lysozyme and RNase A became fully DMF soluble by POXylation. ► POXylated enzymes partly retain their activity in water. The use of enzymes in organic solvents offers a great opportunity for the synthesis...
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Published in: | Journal of biotechnology 2012-06, Vol.159 (3), p.195-203 |
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creator | Konieczny, Stefan Fik, Christoph P. Averesch, Nils J.H. Tiller, Joerg C. |
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► POXylation is successful via pyromellitic acid dianhydride. ► Hen-egg white lysozyme and RNase A became fully DMF soluble by POXylation. ► POXylated enzymes partly retain their activity in water.
The use of enzymes in organic solvents offers a great opportunity for the synthesis of complex organic compounds and is therefore in focus of current research. In this work we describe the synthesis of poly(2-methyl-1,3-oxazoline) (PMOx) and poly(2-ethyl-1,3-oxazoline) (PEtOx) enzyme conjugates with hen-egg white lysozyme, RNase A and α-chymotrypsin using a new coupling technique. The POXylation was carried out reacting pyromellitic acid dianhydride subsequently with ethylenediamine terminated POx and then with the NH2-groups of the respective enzymes. Upon conjugation with the polymers, RNase A and lysozyme became fully soluble in DMF (1.4mg/ml). These are the first examples of fully POXylated proteins, which become organosoluble. The synthesized enzyme conjugates were characterized by SDS-PAGE, isoelectric focusing, dynamic light scattering and size exclusion chromatography, which all indicated the full POXylation of the enzymes. The modified enzymes even partly retained their activity in water. With α-chymotrypsin as example we could demonstrate that the molecular weight of the attached polymer significantly influences the activity. |
doi_str_mv | 10.1016/j.jbiotec.2012.01.016 |
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► POXylation is successful via pyromellitic acid dianhydride. ► Hen-egg white lysozyme and RNase A became fully DMF soluble by POXylation. ► POXylated enzymes partly retain their activity in water.
The use of enzymes in organic solvents offers a great opportunity for the synthesis of complex organic compounds and is therefore in focus of current research. In this work we describe the synthesis of poly(2-methyl-1,3-oxazoline) (PMOx) and poly(2-ethyl-1,3-oxazoline) (PEtOx) enzyme conjugates with hen-egg white lysozyme, RNase A and α-chymotrypsin using a new coupling technique. The POXylation was carried out reacting pyromellitic acid dianhydride subsequently with ethylenediamine terminated POx and then with the NH2-groups of the respective enzymes. Upon conjugation with the polymers, RNase A and lysozyme became fully soluble in DMF (1.4mg/ml). These are the first examples of fully POXylated proteins, which become organosoluble. The synthesized enzyme conjugates were characterized by SDS-PAGE, isoelectric focusing, dynamic light scattering and size exclusion chromatography, which all indicated the full POXylation of the enzymes. The modified enzymes even partly retained their activity in water. With α-chymotrypsin as example we could demonstrate that the molecular weight of the attached polymer significantly influences the activity.</description><identifier>ISSN: 0168-1656</identifier><identifier>EISSN: 1873-4863</identifier><identifier>DOI: 10.1016/j.jbiotec.2012.01.016</identifier><identifier>PMID: 22306109</identifier><language>eng</language><publisher>Netherlands: Elsevier B.V</publisher><subject>Animals ; Benzoates - chemistry ; Biotechnology ; Cattle ; Chickens ; Chromatography, Gel ; Electrophoresis, Polyacrylamide Gel ; Enzyme-conjugate ; gel chromatography ; Hydrolases - chemistry ; Hydrolases - metabolism ; Isoelectric Focusing ; light scattering ; lysozyme ; Molecular Weight ; Organic solvent ; Oxazoles - chemistry ; polyacrylamide gel electrophoresis ; Polyamines - chemistry ; polymers ; Polyoxazoline ; proteins ; Pyromellitic acid dianhydride ; ribonucleases ; Solubility ; solvents</subject><ispartof>Journal of biotechnology, 2012-06, Vol.159 (3), p.195-203</ispartof><rights>2012 Elsevier B.V.</rights><rights>Copyright © 2012 Elsevier B.V. All rights reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c422t-e53f05a27b05106c1ced2a4f5dd14f7358defc7ed39fe62ca464d1eef315d5543</citedby><cites>FETCH-LOGICAL-c422t-e53f05a27b05106c1ced2a4f5dd14f7358defc7ed39fe62ca464d1eef315d5543</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/22306109$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Konieczny, Stefan</creatorcontrib><creatorcontrib>Fik, Christoph P.</creatorcontrib><creatorcontrib>Averesch, Nils J.H.</creatorcontrib><creatorcontrib>Tiller, Joerg C.</creatorcontrib><title>Organosoluble enzyme conjugates with poly(2-oxazoline)s via pyromellitic acid dianhydride</title><title>Journal of biotechnology</title><addtitle>J Biotechnol</addtitle><description>[Display omitted]
► POXylation is successful via pyromellitic acid dianhydride. ► Hen-egg white lysozyme and RNase A became fully DMF soluble by POXylation. ► POXylated enzymes partly retain their activity in water.
The use of enzymes in organic solvents offers a great opportunity for the synthesis of complex organic compounds and is therefore in focus of current research. In this work we describe the synthesis of poly(2-methyl-1,3-oxazoline) (PMOx) and poly(2-ethyl-1,3-oxazoline) (PEtOx) enzyme conjugates with hen-egg white lysozyme, RNase A and α-chymotrypsin using a new coupling technique. The POXylation was carried out reacting pyromellitic acid dianhydride subsequently with ethylenediamine terminated POx and then with the NH2-groups of the respective enzymes. Upon conjugation with the polymers, RNase A and lysozyme became fully soluble in DMF (1.4mg/ml). These are the first examples of fully POXylated proteins, which become organosoluble. The synthesized enzyme conjugates were characterized by SDS-PAGE, isoelectric focusing, dynamic light scattering and size exclusion chromatography, which all indicated the full POXylation of the enzymes. The modified enzymes even partly retained their activity in water. With α-chymotrypsin as example we could demonstrate that the molecular weight of the attached polymer significantly influences the activity.</description><subject>Animals</subject><subject>Benzoates - chemistry</subject><subject>Biotechnology</subject><subject>Cattle</subject><subject>Chickens</subject><subject>Chromatography, Gel</subject><subject>Electrophoresis, Polyacrylamide Gel</subject><subject>Enzyme-conjugate</subject><subject>gel chromatography</subject><subject>Hydrolases - chemistry</subject><subject>Hydrolases - metabolism</subject><subject>Isoelectric Focusing</subject><subject>light scattering</subject><subject>lysozyme</subject><subject>Molecular Weight</subject><subject>Organic solvent</subject><subject>Oxazoles - chemistry</subject><subject>polyacrylamide gel electrophoresis</subject><subject>Polyamines - chemistry</subject><subject>polymers</subject><subject>Polyoxazoline</subject><subject>proteins</subject><subject>Pyromellitic acid dianhydride</subject><subject>ribonucleases</subject><subject>Solubility</subject><subject>solvents</subject><issn>0168-1656</issn><issn>1873-4863</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2012</creationdate><recordtype>article</recordtype><recordid>eNqNkU9v1DAQxS0EokvhIwA5tocsHv9L9oRQRQGpUg_QAyfLa4-3jpJ4sZNC-unxaheuII000uj3Zp7eEPIa6BooqHfdutuGOKFdMwpsTaGUekJW0Da8Fq3iT8mqTNoalFRn5EXOHaVUbCQ8J2eMcaqAblbk-23amTHm2M_bHiscH5cBKxvHbt6ZCXP1M0z31T72ywWr4y_zGPsw4mWuHoKp9kuKA_Z9mIKtjA2ucsGM94tLweFL8sybPuOrUz8nd9cfv119rm9uP325-nBTW8HYVKPknkrDmi2VQJUFi44Z4aVzIHzDZevQ2wYd33hUzBqhhANEz0E6KQU_JxfHvfsUf8yYJz2EbIsrM2KcswbKaCtaUPw_UOCNarg4oPKI2hRzTuj1PoXBpKVAB07pTp8eoA8P0BRKqaJ7czoxbwd0f1V_Ei_A2yPgTdRml0LWd1_LBkkPctlAId4fCSypPQRMOtuAY8klJLSTdjH8w8Rv62qjxQ</recordid><startdate>20120615</startdate><enddate>20120615</enddate><creator>Konieczny, Stefan</creator><creator>Fik, Christoph P.</creator><creator>Averesch, Nils J.H.</creator><creator>Tiller, Joerg C.</creator><general>Elsevier B.V</general><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><scope>7QO</scope><scope>8FD</scope><scope>FR3</scope><scope>P64</scope></search><sort><creationdate>20120615</creationdate><title>Organosoluble enzyme conjugates with poly(2-oxazoline)s via pyromellitic acid dianhydride</title><author>Konieczny, Stefan ; Fik, Christoph P. ; Averesch, Nils J.H. ; Tiller, Joerg C.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c422t-e53f05a27b05106c1ced2a4f5dd14f7358defc7ed39fe62ca464d1eef315d5543</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2012</creationdate><topic>Animals</topic><topic>Benzoates - chemistry</topic><topic>Biotechnology</topic><topic>Cattle</topic><topic>Chickens</topic><topic>Chromatography, Gel</topic><topic>Electrophoresis, Polyacrylamide Gel</topic><topic>Enzyme-conjugate</topic><topic>gel chromatography</topic><topic>Hydrolases - chemistry</topic><topic>Hydrolases - metabolism</topic><topic>Isoelectric Focusing</topic><topic>light scattering</topic><topic>lysozyme</topic><topic>Molecular Weight</topic><topic>Organic solvent</topic><topic>Oxazoles - chemistry</topic><topic>polyacrylamide gel electrophoresis</topic><topic>Polyamines - chemistry</topic><topic>polymers</topic><topic>Polyoxazoline</topic><topic>proteins</topic><topic>Pyromellitic acid dianhydride</topic><topic>ribonucleases</topic><topic>Solubility</topic><topic>solvents</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Konieczny, Stefan</creatorcontrib><creatorcontrib>Fik, Christoph P.</creatorcontrib><creatorcontrib>Averesch, Nils J.H.</creatorcontrib><creatorcontrib>Tiller, Joerg C.</creatorcontrib><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><collection>Biotechnology Research Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><jtitle>Journal of biotechnology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Konieczny, Stefan</au><au>Fik, Christoph P.</au><au>Averesch, Nils J.H.</au><au>Tiller, Joerg C.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Organosoluble enzyme conjugates with poly(2-oxazoline)s via pyromellitic acid dianhydride</atitle><jtitle>Journal of biotechnology</jtitle><addtitle>J Biotechnol</addtitle><date>2012-06-15</date><risdate>2012</risdate><volume>159</volume><issue>3</issue><spage>195</spage><epage>203</epage><pages>195-203</pages><issn>0168-1656</issn><eissn>1873-4863</eissn><abstract>[Display omitted]
► POXylation is successful via pyromellitic acid dianhydride. ► Hen-egg white lysozyme and RNase A became fully DMF soluble by POXylation. ► POXylated enzymes partly retain their activity in water.
The use of enzymes in organic solvents offers a great opportunity for the synthesis of complex organic compounds and is therefore in focus of current research. In this work we describe the synthesis of poly(2-methyl-1,3-oxazoline) (PMOx) and poly(2-ethyl-1,3-oxazoline) (PEtOx) enzyme conjugates with hen-egg white lysozyme, RNase A and α-chymotrypsin using a new coupling technique. The POXylation was carried out reacting pyromellitic acid dianhydride subsequently with ethylenediamine terminated POx and then with the NH2-groups of the respective enzymes. Upon conjugation with the polymers, RNase A and lysozyme became fully soluble in DMF (1.4mg/ml). These are the first examples of fully POXylated proteins, which become organosoluble. The synthesized enzyme conjugates were characterized by SDS-PAGE, isoelectric focusing, dynamic light scattering and size exclusion chromatography, which all indicated the full POXylation of the enzymes. The modified enzymes even partly retained their activity in water. With α-chymotrypsin as example we could demonstrate that the molecular weight of the attached polymer significantly influences the activity.</abstract><cop>Netherlands</cop><pub>Elsevier B.V</pub><pmid>22306109</pmid><doi>10.1016/j.jbiotec.2012.01.016</doi><tpages>9</tpages></addata></record> |
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subjects | Animals Benzoates - chemistry Biotechnology Cattle Chickens Chromatography, Gel Electrophoresis, Polyacrylamide Gel Enzyme-conjugate gel chromatography Hydrolases - chemistry Hydrolases - metabolism Isoelectric Focusing light scattering lysozyme Molecular Weight Organic solvent Oxazoles - chemistry polyacrylamide gel electrophoresis Polyamines - chemistry polymers Polyoxazoline proteins Pyromellitic acid dianhydride ribonucleases Solubility solvents |
title | Organosoluble enzyme conjugates with poly(2-oxazoline)s via pyromellitic acid dianhydride |
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