超越折叠酶:具有氧活性的"固体伴侣"释放可回收的,无HPLC氧化蛋白折叠
Shunpei Iwamoto1, Yuya Nishizawa1, Hayato Yokose2,3
1Department of Chemistry, School of Science, Tokai University, 4-1-1 Kitakaname, Hiratsuka-Shi, Kanagawa 259-1292, Japan.
JACS Au
|February 27, 2026
概括
研究人员开发了一种可重复使用的聚合物"固体伴侣",模仿蛋白质二硫化异合酶 (PDI) 进行高效的氧化蛋白折叠. 这种新型材料使得可回收,无酶的二硫化物键形成和蛋白质重新折叠成为可能,克服了当前方法的局限性.
科学领域:
- 生物化学 生物化学
- 聚合物化学 聚合物化学
- 材料科学 材料科学 材料科学
背景情况:
- 氧化蛋白折叠对蛋白质功能至关重要,通常由蛋白质二硫化异构酶 (PDI) 催化.
- 目前的PDI应用受到酶不稳定性,一次性使用要求和同质系统限制的限制.
- 现有的人工PDI模仿剂缺乏足够的氧化还原活性和聚合抑制,无法进行强大的体外折叠.
研究的目的:
- 开发一种可回收的固体相系统,模仿PDI在氧化蛋白折叠中的双重活性.
- 为了创建一个强大的替代酶用于in vitro蛋白质重新折叠和二硫化物键形成.
- 展示工程聚合物材料对复杂酶功能的潜力.
主要方法:
- 在聚钢珠上对循环脱化物进行共价固定,以创建一个氧化还原活性,疏水的接口.
- 利用固相材料作为蛋白质捕获和氧化折叠的"固体伴侣".
- 评估二硫化键形成和异构化的催化活性,并评估可重复使用性和聚合抑制.
主要成果:
- 基于聚合物的固体伴侣在非生物表面上表现出类似PDI的双重活性.
- 在本地和蛋白质生产中达到高达99%的产量.
- 在多个重复使用周期中保持完整的催化活性,性能优于同质系统.
- 有效抑制蛋白质聚合,即使在高基质度.
结论:
- 工程耐用的聚合物材料可以复制复杂的酶功能,如氧化折叠.
- 开发的固体相策略可以实现可回收,无酶的氧化折叠,克服自然PDI的局限性.
- 这种方法建立了一个范式,用于将酶行为转化为具有工业潜力的可扩展合成系统.
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