SGER: Investigation of dynamic actions of cellulolytic enzymes with micro-cantilever sensors

CBET-0843921 XI本研究将促进对纤维素酶在非均相体系中不溶于水的天然底物上的动态作用的机理的理解。纤维素酶与植物细胞结构和层次之间的相互作用，如纤维素、半纤维素和木质素结构，目前还知之甚少。这可以部分归因于缺乏能够检查酶和固体底物之间的界面相互作用的分析工具。一些现有的技术，如椭偏仪和石英晶体微天平，最近已被应用于此类研究，但无法达到解剖整个纤维素分解过程所需的灵敏度。由于纤维素酶是降解纤维素生物质的关键酶，对纤维素酶机理的缺乏严重阻碍了利用纤维素酶生产生物燃料的研究进展。一种新的方法将被用来研究纤维素酶与水不溶结晶纤维素底物之间的界面相互作用，该方法利用一种新兴的微悬臂梁技术。这种方法将使我们能够对纤维素酶的作用进行更定量的评估，并潜在地建立酶和底物的结构-功能关系，这将是开发具有更高效率和稳定性的用于生物处理的新纤维素酶的关键。更好地了解这些相互作用也将有助于我们从总体上了解界面酶催化的基础知识，更重要的是将加快以纤维素生物质为原料的生物燃料的开发，作为一种可能对国民经济、环境和国家安全产生深远影响的替代能源。这项拟议的研究需要化学家、生物学家、工程师和物理学家共同努力，采取多学科的方法。这将促进德雷塞尔大学发展高度跨学科的研究和教育环境的持续努力，这将为研究生和本科生提供一个接触广泛科学领域的绝佳机会，并为他们在未来的科学职业生涯中解决复杂问题做好准备。这项工作的结果将在相关会议上公布，并在同行评议的期刊上发表。

CBET-0843921XiThis research will advance the mechanistic understanding of dynamic actions of cellulolytic enzyme cellulases on water-insoluble native substrates in a heterogeneous system. The interactions between cellulases and plant cell architecture and hierarchy, such as cellulose, hemicellulose, and lignin structures are currently poorly understood. This can be attributed partially to the lack of an analytical tool capable of examining the interfacial interactions between the enzymes and the solid substrates. Some of the existing technologies such as Ellipsometry and Quartz Crystal Microbalance have recently been applied in such studies but unable to achieve the sensitivity required for dissecting the entire cellulolytic processes. Because cellulolytic enzymes are the key enzymes in degrading the cellulosic biomass, the lack of mechanistic understanding of cellulases has significantly hindered the progress of developing an efficient and economic bioprocess to produce biofuel from cellulosic biomass. A novel method will be used to investigate the interfacial interactions between cellulases and water-insoluble crystalline cellulose substrates by taking advantage of an emerging micro-cantilever technology. This approach will allow us to obtain a more quantitative assessment of cellulase actions and potentially establish structural-functional relationships of both enzymes and substrates, which will be essential for developing a new cellulase with improved efficiency and stability for bioprocessing. A better understanding of these interactions will also contribute to our fundamental knowledge of interfacial enzyme catalysis in general, and more importantly will accelerate the development of biofuels from cellulosic biomass as an alternative energy source that could potentially have a profound impact on national economy, environment, and national security. The proposed research requires a multidisciplinary approach with a collaborative effort of chemists, biologists, engineer, and physicists. This will facilitate the on-going effort of developing a highly interdisciplinary research and educational environment at Drexel University, which will potentially offer a great opportunity for both graduate and undergraduate students to be exposed to a broad range of scientific fields, and prepare them to tackle the complex problems in their future scientific career. The results of this work will be presented at relevant conferences, and published in peer-reviewed journals.