| 一种温敏性水凝胶复性体系:聚N-异丙基丙烯酰胺的制备及其在溶菌酶复性中的应用 |
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| 引用本文： | 崔志芳,关怡新,姚善泾. 一种温敏性水凝胶复性体系:聚N-异丙基丙烯酰胺的制备及其在溶菌酶复性中的应用[J]. 中国化学工程学报, 2004, 12(4) |
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| 作者姓名： | 崔志芳 关怡新 姚善泾 |
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| 摘 要： |
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A Temperature-sensitive Hydrogel Refolding System:Preparation of Poly(N-isopropyl acrylamide) and Its Application in Lysozyme Refolding |
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| CUI Zhifang,Guan Yixin,YAO Shanjing. A Temperature-sensitive Hydrogel Refolding System:Preparation of Poly(N-isopropyl acrylamide) and Its Application in Lysozyme Refolding[J]. Chinese Journal of Chemical Engineering, 2004, 12(4) |
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| Authors: | CUI Zhifang Guan Yixin YAO Shanjing |
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| Affiliation: | Department of Chemical and Biochemical Engineering, Zhejiang University, Hangzhou 310027, China;Department of Chemical and Biochemical Engineering, Zhejiang University, Hangzhou 310027, China;Department of Chemical and Biochemical Engineering, Zhejiang University, Hangzhou 310027, China |
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| Abstract: | Temperature-sensitive hydrogel-poly(N-isopropyl acrylamide) (PNIPA) was prepared and applied to protein refolding. PNIPA gel disks and gel particles were synthesized by the solution polymerization and inverse suspension polymerization respectively. The swelling kinetics of the gels was also studied. With these prepared PNIPA gels, the model protein lysozyme was renatured. Within 24 h, PNIPA gel disks improved the yield of lysozyme activity by 49.3% from 3375.2 U.mg-1 to 5038.8 U.mg-1. With the addition of faster response PNIPA gel beads,the total lysozyme activity recovery was about 68.98% in 3h, as compared with 42.03% by simple batch dilution.The novel refolding system with PNIPA enables efficient refolding especially at high protein concentrations. Discussion about the mechanism revealed that when PNIPA gels were added into the refolding buffer, the hydrophobic interactions between denatured proteins and polymer gels could prevent the aggregation of refolding intermediates,thus enhanced the protein renaturation. |
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| Keywords: | poly(N-isopropyl acrylamide) hydrogel free radical polymerization protein refolding lysozyme |
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