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

本研究将促进对纤维素酶在非均相体系中对水不溶性天然底物的动态作用的机理理解。纤维素酶与植物细胞结构和层次（如纤维素、半纤维素和木质素结构）之间的相互作用目前知之甚少。这可以部分归因于缺乏能够检查酶和固体基质之间的界面相互作用的分析工具。一些现有的技术，如椭圆偏振法和石英晶体微量天平最近已被应用在这样的研究，但无法实现所需的灵敏度解剖整个纤维素分解过程。由于纤维素分解酶是降解纤维素类生物质的关键酶，因此对纤维素酶的机理缺乏了解已经显著阻碍了开发从纤维素类生物质生产生物燃料的高效且经济的生物过程的进展。利用微悬臂梁技术研究了纤维素酶与水不溶性结晶纤维素基质之间的界面相互作用。这种方法将使我们能够获得一个更定量的评估纤维素酶的行动和潜在的建立酶和底物的结构-功能关系，这将是必不可少的开发一种新的纤维素酶，提高效率和稳定性的生物加工。更好地了解这些相互作用也将有助于我们对界面酶催化的基本知识，更重要的是将加速从纤维素生物质中开发生物燃料作为替代能源，可能对国民经济，环境和国家安全产生深远影响。拟议的研究需要化学家，生物学家，工程师和物理学家的合作努力的多学科方法。这将促进在德雷克塞尔大学发展高度跨学科研究和教育环境的持续努力，这将可能为研究生和本科生提供一个很好的机会，让他们接触到广泛的科学领域，并为他们在未来的科学生涯中解决复杂的问题做好准备。这项工作的结果将在有关会议上提出，并发表在同行评审的期刊上。

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.