Molecular mechanisms of catalysis and the basis of substrate specificity in polysaccharide hydrolases

多糖水解酶催化的分子机制及底物特异性的基础。沿着水解途径的反应中间体和大麦b -葡聚糖外水解酶和内水解酶的底物特异性的分子决定因素将使用晶体学和动力学分析来定义。这些酶在高等植物发育过程中的细胞壁代谢和植物与病原体的相互作用中起着至关重要的作用。项目目标的实现不仅将提供催化机制的基本信息，而且还将提供操纵酶特异性的机会。此外，酶的定点诱变将用于产生糖合酶，将评估它们合成新的低聚糖和多糖产物的能力，其中一些可能显示免疫调节活性。

Molecular mechanisms of catalysis and the basis of substrate specificity in polysaccharide hydrolases. Reaction intermediates along hydrolytic pathways and molecular determinants of substrate specificity of barley B-glucan exo- and endohydrolases will be defined using crystallographic and kinetic analyses. These enzymes are of central importance in cell wall metabolism during development of higher plants, and in plant-pathogen interactions. Realization of the project objectives will not only provide fundamental information on catalytic mechanisms, but will also provide opportunities to manipulate enzyme specificity. Further, site-directed mutagenesis of the enzymes will be used to generate glycosynthases, which will be evaluated for their ability to synthesise novel oligosaccharide and polysaccharide products, some of which might show immunomodulating activity.