One-step immobilization and purification of his-tagged enzyme using poly(2-acetamidoacrylic acid) hydrogel
DC Field | Value | Language |
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dc.contributor.author | Ha, Eun-Ju | - |
dc.contributor.author | Kim, Kyeong Kyu | - |
dc.contributor.author | Park, Hyung Soon | - |
dc.contributor.author | Lee, Sun-Gu | - |
dc.contributor.author | Lee, Jang-Oo | - |
dc.contributor.author | An, Seong Soo A. | - |
dc.contributor.author | Paik, Hyun-Jong | - |
dc.date.available | 2020-02-29T00:46:18Z | - |
dc.date.created | 2020-02-06 | - |
dc.date.issued | 2013-01 | - |
dc.identifier.issn | 1598-5032 | - |
dc.identifier.uri | https://scholarworks.bwise.kr/gachon/handle/2020.sw.gachon/14843 | - |
dc.description.abstract | Ni2+-Complexed poly(2-acetamidoacrylic acid) (PAAA) hydrogel support was developed for the one-step immobilization and purification of recombinant histidine-tagged glutamyl aminopeptidase (His-tagged GAP). Ni2+-PAAA hydrogel was prepared from the polymerization of 2-acetamidoacrylic acid (AAA) and 2,2-[(1,4-dioxo-1,4-butanediyl) diamino] bis(2-propenoic acid) (DBDBPA) with potassium persulfate in dimethyl sulfoxide (DMSO), followed by Ni(2+)complexation. His-tagged GAP was immobilized directly from the cell lysate onto the Ni2+-PAAA hydrogel support and then purified. Catalytic activity of immobilized His-tagged GAP for the hydrolysis of alanylpara-nitroanilide revealed 90% conversion after 30 min of incubation, indicating sustained catalytic activity. The hydrogel-immobilized enzyme also exhibited enhanced thermal stability of sustained 70% activity after 1 h incubation at 60 A degrees C, while the free enzyme activity was reduced to 50% at the same condition. After four cycles of hydrogel regeneration, the immobilized enzyme lost only 20% of its initial activity. Ni2+-PAAA hydrogel provided a new and convenient immobilization/purification system for His-tag enzymes through easy and simple procedures. | - |
dc.language | 영어 | - |
dc.language.iso | en | - |
dc.publisher | POLYMER SOC KOREA | - |
dc.relation.isPartOf | MACROMOLECULAR RESEARCH | - |
dc.subject | LIPASE | - |
dc.subject | BEADS | - |
dc.subject | ADSORPTION | - |
dc.subject | PROTEINS | - |
dc.subject | MATRICES | - |
dc.title | One-step immobilization and purification of his-tagged enzyme using poly(2-acetamidoacrylic acid) hydrogel | - |
dc.type | Article | - |
dc.type.rims | ART | - |
dc.description.journalClass | 1 | - |
dc.identifier.wosid | 000314280900002 | - |
dc.identifier.doi | 10.1007/s13233-013-1007-8 | - |
dc.identifier.bibliographicCitation | MACROMOLECULAR RESEARCH, v.21, no.1, pp.5 - 9 | - |
dc.identifier.kciid | ART001738231 | - |
dc.identifier.scopusid | 2-s2.0-84873413560 | - |
dc.citation.endPage | 9 | - |
dc.citation.startPage | 5 | - |
dc.citation.title | MACROMOLECULAR RESEARCH | - |
dc.citation.volume | 21 | - |
dc.citation.number | 1 | - |
dc.contributor.affiliatedAuthor | An, Seong Soo A. | - |
dc.type.docType | Article | - |
dc.subject.keywordAuthor | poly(2-acetamidoacrylic acid) hydrogel | - |
dc.subject.keywordAuthor | purification | - |
dc.subject.keywordAuthor | immobilization | - |
dc.subject.keywordAuthor | his-tagged enzyme | - |
dc.subject.keywordPlus | LIPASE | - |
dc.subject.keywordPlus | BEADS | - |
dc.subject.keywordPlus | ADSORPTION | - |
dc.subject.keywordPlus | PROTEINS | - |
dc.subject.keywordPlus | MATRICES | - |
dc.relation.journalResearchArea | Polymer Science | - |
dc.relation.journalWebOfScienceCategory | Polymer Science | - |
dc.description.journalRegisteredClass | scie | - |
dc.description.journalRegisteredClass | scopus | - |
dc.description.journalRegisteredClass | kci | - |
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