CAREER: Computational Enzymology of Non-Aqueous Biocatalysis: Application to Biomass Pretreatment

1150596-Pfaendtner社会从可持续来源设计和生产化学品和燃料的能力取决于未来生物加工的改进。 非水溶剂如离子液体可能会彻底改变生物工艺的效率和经济性，特别是在用于发酵的生物质的酶促预处理中。 非水溶剂赋予生物催化剂改进或以前未知的性能的潜力是令人兴奋的，并可能改变其在工业中的应用。 尽管这一伟大的承诺，溶剂，酶的结构和反应性之间的关系的不完全的理解，阻碍了重大的科学技术的发展，在这个竞技场。 具体而言，缺乏的是将溶剂的分子特征与其对酶稳定性和活性的影响相关联的能力。 在溶剂降低酶活性的情况下，与其自然环境相比，我们没有系统地了解活性丧失的根本原因或如何有效地使用诱变技术来克服活性丧失的工具。这项由华盛顿大学的Jim Pfaendtner提出的CAREER项目将开发和验证一种新的多功能基因工程。大规模计算建模工具包，用于发现和优化生物过程溶剂环境。 该研究计划的目标是研究两种糖苷水解酶（GH）与三种离子液体（IL）的组合。 这些酶广泛用于生物加工应用，选择IL是因为它们的成本效益和用作生物质增溶剂的潜力。 利用先进的模拟技术，PI和他的团队将系统地确定IL改变GH酶活性的潜在分子尺度机制。 正在追求四个研究目标，以区分活动变化是基于结构、动态还是两者的结合。该项目产生了几个更广泛的技术影响。 研究的成功完成将导致新的生物质利用技术，可以消除一个或多个预处理阶段。 此外，这项工作将为研究非水环境中的酶创造一个通用的计算方法。 这种新方法提供了一种新的预测能力，将加速科学发现，降低实验成本，并改变我们使用计算机指导生物过程设计和优化的方式。 与拟议的研究相结合的是一项广泛的外联、教育和培训计划。 教育组件包括一个模拟模块的AP化学类在西雅图高中和一个新的生物催化模块的高级本科实验室课程。 研究培训部分包括为当地高中教师提供研究机会，并与校园外展组织路易斯·斯托克斯少数民族参与联盟（LSAMP）合作，让代表性不足的少数民族参与PI研究小组。 该职业奖由化学，生物工程，环境和运输系统部门的催化和生物催化计划以及化学部门的化学催化计划共同赞助。

1150596-Pfaendtner Societal ability to design and produce chemicals and fuels from sustainable sources relies on future improvements in bioprocessing. Non-aqueous solvents such as ionic liquids may potentially revolutionize bioprocess efficiency and economics, particularly in the enzymatic pretreatment of biomass for fermentation. The potential for non-aqueous solvents to endow biological catalysts with improved or previously unknown properties is exciting and could transform their use in industry. In spite of this great promise, the incomplete understanding of the relationship between solvent, enzyme structure, and reactivity has hampered significant scientific technological development in this arena. Specifically, what is lacking is the ability to relate molecular features of the solvent to its effect on enzyme stability and activity. In cases where solvents lower the activity of the enzyme compared with its natural environment we do not have the tools to systematically understand the root cause of the loss of activity or how to effectively use mutagenesis techniques to overcome the activity loss.This CAREER project put forward By Jim Pfaendtner of the University of Washington will develop and validate a new multi-scale computational modeling toolkit for the discovery and optimization of bioprocess solvent environments. The target of the research proposal is the investigation of two glycoside hydrolase (GH) enzymes with a combination of three ionic liquids (ILs). These enzymes are widely used in bioprocessing applications and the ILs are chosen for their cost effectiveness and potential for use as biomass solubilization agents. Using advanced simulation techniques the PI and his group will systematically determine the underlying molecular scale mechanisms by which ILs change the activity of GH enzymes. Four research objectives are being pursued that will discriminate whether the activity changes are based on structure, dynamics or a combination of both. There are several technological broader impacts stemming from this project. Successful completion of the research will lead to new biomass utilization technologies that could eliminate one or more pretreatment stages. Additionally, this work will create a general computational approach for studying enzymes in non-aqueous environments. The new approach offers a new level of predictive capability that will speed scientific discovery, reduce experimental costs, and transform the way we use computers to guide bioprocess design and optimization. Integrated with the proposed research is an extensive outreach, education and training plan. Educational components include a simulation module for an AP chemistry class at a Seattle High School and a new biocatalysis module for the senior undergraduate laboratory course. Research training components include research opportunities for local High School teachers and collaboration with a campus outreach organization, Louis Stokes Alliance for Minority Participation, (LSAMP) to involve under-represented minorities in the PIs research group. This CAREER award is jointly made and sponsored by the Catalysis and Biocatalysis Program of the Chemical, Bioengineering, Environmental, and Transport Systems Division and the Chemical Catalysis Program of the Division of Chemistry.