One-step immobilization and purification of his-tagged enzyme using poly(2-acetamidoacrylic acid) hydrogel
- Authors
- Ha, Eun-Ju; Kim, Kyeong Kyu; Park, Hyung Soon; Lee, Sun-Gu; Lee, Jang-Oo; An, Seong Soo A.; Paik, Hyun-Jong
- Issue Date
- Jan-2013
- Publisher
- POLYMER SOC KOREA
- Keywords
- poly(2-acetamidoacrylic acid) hydrogel; purification; immobilization; his-tagged enzyme
- Citation
- MACROMOLECULAR RESEARCH, v.21, no.1, pp.5 - 9
- Journal Title
- MACROMOLECULAR RESEARCH
- Volume
- 21
- Number
- 1
- Start Page
- 5
- End Page
- 9
- URI
- https://scholarworks.bwise.kr/gachon/handle/2020.sw.gachon/14843
- DOI
- 10.1007/s13233-013-1007-8
- ISSN
- 1598-5032
- 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.
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Collections - 바이오나노대학 > 바이오나노학과 > 1. Journal Articles
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