d酶固定化蛋白质工程.ppt

Understanding enzyme immobilisation Enzymes are versatile catalysts in the laboratory and on an industrial scale. To broaden their applicability in the laboratory and to ensure their (re)use in manufacturing the stability of enzymes can often require improvement. Immobilisation can address the issue of enzymatic instability. Immobilisation can also help to enable the employment of enzymes in different solvents, at extremes of pH and temperature and exceptionally high substrate concentrations. At the same time substrate-specificity, enantioselectivity and reactivity can be modified. However, most often the molecular and physical-chemical bases of these phenomena have not been elucidated yet. Recent advances in immobilized enzymatic reactors and their applications in proteome analysis Immobilized enzymatic reactors recently have drawn much attention because of the striking advantages, such as high substrate turnover rate and ease in coupling with the separation and detection systems. Carrier materials, which have great effects on the development of the immobilized enzymatic reactors, have always being the focus of study. In this paper, the contributions, mainly in the last 5 years, on the enzymatic reactors and their applications in proteome study are reviewed, with some newly developed inorganic and organic carriers for enzyme immobilization described in details. Moreover, the hyphenation of immobilized enzymatic reactors with the separation and identification systems is also summarized. By reviewing these achievements, it could be seen that enzymatic reactors have very bright future, especially in proteome analysis 固定化酶的研究内容 固定化方法 载体 酶学性质——固定化酶的特性 稳定性研究、改进 最适温度 最适pH 底物特异性 固定化酶的制备方法（一） 吸附法 1． 物理吸附：利用各种固体吸附剂将酶或含酶菌体吸附在其表面上。  选择载体的原则 （1）要有巨大的比表面积 （2） 要有活泼的表面（3） 便于装柱进行连续反应。 优点： 固定化时酶分子的构象很少 或基本不发生变化。 缺点： 结合力弱，易解吸附。 载体： 纤维素、琼脂糖、活性炭、 沸石及硅胶等。 （二）结合法 1 离子键结合法 酶分子 含有离子交换基团的固相载体 常用载体:DEAE-纤维素， DEAE-葡聚糖凝胶 使用注意：pH、离子强度、温度 2 共价键结合法 （1）酶分子中可以形成共价键的基团： 游离氨基