Improved renaturation process of aggregated recombinant proteins through the design of hydrated ionic liquids
Improved renaturation process of aggregated recombinant proteins through the design of hydrated ionic liquids
复制标题
通过水合离子液体的设计改进聚集重组蛋白的复性过程
DOI：
10.1016/j.molliq.2023.121440
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发表时间：
2023
影响因子：
6
通讯作者：
Ohno Hiroyuki
中科院分区：
文献类型：
--
作者：
Fujita Kyoko;Kobayashi Kazune;Ito Anna;Yanagisawa Shun;Ichida Kimiyoshi;Takeda Kouta;Nakamura Nobuhumi;Ohno Hiroyuki
大肠杆菌是重组蛋白表达最常用的宿主。然而，由于以高速率获得非活性聚集，并且复性速率仍然低，因此聚集的重组蛋白的处理的改进是关键问题。本研究的目的是评估使用水合离子液体（IL）直接溶解聚集的重组蛋白和诱导复性。组分离子和合适的水含量的选择导致了相当大的改善，在溶解的聚集蛋白质，其次是重折叠行为。与磷酸二氢阴离子耦合的丁基铵或磷阳离子被确定为适当的离子对，显示出高溶解度，并有助于再折叠。柱方法显示出巨大的潜力，在过渡的溶解和复性的蛋白质在水合IL到缓冲溶液中，实现IL去除和蛋白质纯化。聚集蛋白的成功回收的活性水平表明，该方法是一种有前途的方法，用于开发新的治疗聚集的重组蛋白。
Escherichia coliis the most commonly used host for recombinant protein expression. However, since inactive aggregation is obtained at a high rate and the renaturation rate is still low, the improvement of the treatment of aggregated recombinant proteins is a critical issue. This study aimed to evaluate the use of hydrated ionic liquids (ILs) to directly dissolve aggregated recombinant proteins and induce refolding. Selection of component ions and suitable water content resulted in a considerable improvement in the dissolution of the aggregated protein, followed by refolding behaviour. Ammonium or phosphonium cations with butyl groups coupled with dihydrogen phosphate anions were identified as appropriate ion pairs, showing high solubility and contributing to refolding. The column method showed great potential in the transition of dissolved and refolded proteins in hydrated ILs into the buffer solution, achieving both IL removal and protein purification. The successfully recovered activity levels of aggregated proteins indicate that this method is a promising approach for the development of novel treatments for aggregated recombinant proteins.
灰鬼伞 (Coprinopsis cinerea) 糖苷水解酶家族 6 酶的表征
DOI：
10.1271/bbb.80888
发表时间：
2009
期刊：
Bioscience, Biotechnology, and Biochemistry
影响因子：
--
作者：
Yuan Liu;K. Igarashi;S. Kaneko;T. Tonozuka;M. Samejima;K. Fukuda;Makoto Yoshida
通讯作者：
Makoto Yoshida
影响因子：
6.2
作者：
Fujita, Kyoko;MacFarlane, Douglas R.;Ohno, Hiroyuki
通讯作者：
Ohno, Hiroyuki