Halling Peter J, Ulijn Rein V, Flitsch Sabine L
Department of Pure & Applied Chemistry, University of Strathclyde, Glasgow G1 1XW, UK.
Curr Opin Biotechnol. 2005 Aug;16(4):385-92. doi: 10.1016/j.copbio.2005.06.006.
With increasing interest in automated synthesis and screening protocols, solid supported chemistry and biochemistry are attractive technologies. Studies with surface-immobilised substrates have been carried out to analyse enzyme accessibility, kinetics and thermodynamics. Several interesting new methods have been developed to monitor enzyme action on substrates attached to a solid phase such as polymer beads glass or gold surfaces. These include fluorescence measurements, MALDI-TOF mass spectrometry, and the use of quartz crystal microbalances to measure weight changes of immobilised molecules directly on the surface. Approaches that allow spatial resolution in single beads have also been reported. The ability of enzymes to reach the inside of beads is becoming better characterised and new supports have been developed that allow improved accessibility. The equilibrium position of reactions on the solid surface can be substantially shifted compared with reactions in solution, and this can be usefully exploited using hydrolases in reverse. Research is also starting to tackle the way in which kinetics are modified when the substrates are surface immobilised.
随着对自动化合成和筛选方案的兴趣日益增加，固相支持的化学和生物化学成为颇具吸引力的技术。人们开展了有关表面固定化底物的研究，以分析酶的可及性、动力学和热力学。已经开发出了几种有趣的新方法来监测酶对附着于固相（如聚合物珠、玻璃或金表面）的底物的作用。这些方法包括荧光测量、基质辅助激光解吸电离飞行时间质谱，以及使用石英晶体微天平直接测量表面固定化分子的重量变化。也有报道称在单个珠子中实现空间分辨率的方法。酶进入珠子内部的能力正得到更好的表征，并且已经开发出了能提高可及性的新型载体。与溶液中的反应相比，固体表面上反应的平衡位置可能会发生显著偏移，利用水解酶的逆反应可以有效利用这一点。研究也开始探讨底物表面固定化时动力学是如何改变的。
