由酶不动化促进的不同基底蛋白对 ficin 特异性的变化
Alex D Gonzalez-Vasquez1, El Siar Hocine2, Marcela Urzúa3
1Departamento de Biocatalisis, ICP-CSIC, Campus UAM-CSIC, Madrid 28049, Spain; Departamento de Química, Facultad de Ciencias, Universidad de Chile, Las Palmeras 3425, Casilla 653, Santiago, Ñuñoa 7800003, Chile.
Enzyme and microbial technology
|September 25, 2024
概括
使用氧或阿斯巴拉提克/1,6 六甲基二胺 (Asp/HA) 对阿加珠的酶固定支持改变的 ficin.
科学领域:
- 生物化学 生物化学
- 酶不移动化酶不移动化
- 蛋白质化学 蛋白质化学
背景情况:
- 酶固定对于生物催化剂的可重复使用性和稳定性至关重要.
- 不同的固定支持和方法可以显著影响酶活性和特异性.
- 植物蛋白酶菲辛具有多种应用,但需要优化的固定化策略.
研究的目的:
- 为了比较 ficin 在 glyoxyl-agarose 和 Aspartic/1,6 hexamethylenediamine (Asp/HA) -agarose 珠子上固定化的活性.
- 为了研究甲基修饰对固定 ficin 活性的影响.
- 评估固定对 ficin 对各种蛋白质和基质的特异性的影响.
主要方法:
- 菲辛被固定在氧和Asp/HA-agarose支器上.
- 使用Asp/HA不动化的 ficin 进一步使用甲进行了修改.
- 使用素,血红蛋白,牛血清白蛋白和醇-氨酸-p-nitroanilide在pH 7和5时测定了酶活性.
主要成果:
- 氧基-阿加洛斯的固定不动并没有降低 ficin 的活性,即使反应时间延长.
- Asp/HA 支持诱导了 ficin 活性的显著变化,其中一些基质显示活性增加,而另一些基质则减少.
- 甲基治疗进一步增强了这些活性变化,表明了显著的形状变化和改变的基质特异性.
结论:
- 固定化策略,特别是支持和化学修饰的选择,深深地影响了 ficin 的酶特性.
- 固定化引起的形状变化可能导致基质特异性的改变,影响酶性能.
- 这些发现对酶失活研究和定制生物催化剂的开发有影响.
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