使用alpha/beta-barrel架构的新催化活动的定向演变
M M Altamirano1, J M Blackburn, C Aguayo
1Cambridge Centre for Protein Engineering, and Cambridge University Chemical Laboratory, MRC Centre, UK.
Nature
|February 25, 2000
概括
科学家们使用一种常见的蛋白质结构,即α/β-barrel scaffold,进化出了一种新的酶. 这种定向进化方法创造了一种具有增强特异性的新型基甲酸异相酶,为新的生物催化剂铺平了道路.
科学领域:
- 生物催化和酶工程 生物催化和酶工程
- 蛋白质支架的进化过程
- 酶的有针对性的进化.
背景情况:
- 酶是生物系统中至关重要的生物催化剂,但工业应用需要具有定制功能的新酶.
- 酶进化经常重新利用现有的蛋白质支架来获得新的催化活性.
- 阿尔法/贝塔桶是一种普遍的蛋白质折叠,表明其作为多功能支架的潜力.
研究的目的:
- 通过在现有的蛋白质支架上设计新的催化功能来模仿自然的酶进化.
- 通过使用醇-3-甘-酸盐合成酶α/β-桶脚手架来演变酸化基酸异构酶活性.
- 为了证明定向进化的可行性,从常见的蛋白质折叠中创建新的生物催化剂.
主要方法:
- 在阿尔法/β桶支架内结合了基乙烯酸酸盐的结合部位与异构酶催化模板.
- 使用 in vitro 突变发生和重组以准催化模板.
- 采用体内选择来识别和分离已进化的酶.
主要成果:
- 从-3-甘-酸盐合成酶支架中成功进化出一种具有酸乙烯酸异构酶活性的新型酶.
- 工程酶表现出与自然酶相比的催化特性.
- 新酶的特异性常数比其天然对应物更高.
结论:
- 阿尔法/贝塔桶架构是新生物催化剂定向演变的有希望的平台.
- 这项研究表明,使用常见的蛋白质折叠,酶功能的成功分离进化.
- 这些发现支持了重新利用现有的蛋白质支架来创造新的酶活性的战略.
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