Istituto di Chimica del Riconoscimento Molecolare, CNR, Via Mario Bianco 9, Milano, Italy.
Chem Soc Rev. 2013 Aug 7;42(15):6250-61. doi: 10.1039/c3cs35495d.
Protein conformation plays a crucial role in determining both the catalytic efficiency and the chemo-, regio- and enantioselectivity of enzymes, thus eventually influencing their exploitability in biotechnological applications. Inevitably, immobilisation processes alter the natural molecular environment of enzymes, and quite often affect their catalytic activity through different mechanisms such as reduced accessibility of the substrate to the catalytic active centre, loss of the enzyme dynamic properties and alteration of the conformational integrity of the enzyme. This tutorial review outlines first the most common spectroscopic techniques used for investigating the conformation of immobilized proteins, and then examines how protein loading and polar and hydrophobic/hydrophilic interactions with the carrier affect the structural and dynamic features of enzymes. The nanoscale-level studies in which protein conformational changes, determined either by experimental approaches or by homology modelling, are correlated with the size and shape of the support are also discussed. Altogether, these results should provide useful information on how supports and/or enzymes have to be tailored to improve biocatalyst performance.
蛋白质构象在决定酶的催化效率以及化学、区域和对映选择性方面起着至关重要的作用，因此最终会影响它们在生物技术应用中的可利用性。不可避免的是，固定化过程会改变酶的天然分子环境，并且经常通过不同的机制影响其催化活性，例如底物与催化活性中心的可及性降低、酶的动态特性丧失以及酶构象完整性的改变。本综述首先概述了用于研究固定化蛋白质构象的最常见的光谱技术，然后研究了蛋白质负载以及与载体的极性和疏水性/亲水性相互作用如何影响酶的结构和动态特性。还讨论了通过实验方法或同源建模确定的蛋白质构象变化与载体的大小和形状相关的纳米级研究。总之，这些结果应该为如何调整载体和/或酶以提高生物催化剂性能提供有用的信息。
