Synthetic Biology: Impacts of structural alterations on function in a mobile enzyme sequestration platform (mESP)

该项目由分子和细胞生物科学司的系统和合成生物学组以及生物基础设施司的人力资源组共同资助。对替代运输燃料的需求可以补充或取代化石燃料(例如汽油和柴油)是一个持续的全球关注问题。用于生产生物乙醇和生物柴油的相同的化学转化技术正被用于生产生物燃料。这一过程涉及使用苛刻的化学品(如酸)，这给生产商带来了危险材料的处理和处置问题。以酶为基础的原料转化为生物燃料的危害较小，而且可以说更环保。不幸的是，微生物或非生物酶介导的反应通常不如化学催化那么有效。本项目致力于设计和测试一种在高温和极端pH条件下提高酶效率的试验性酶固定平台系统。这种情况在工业反应过程中很常见。该项目的基础是基础生物化学研究，在该研究中，对流动酶分离平台进行了改进和有效性检查。对这些平台进行了研究，以表征它们在恶劣反应条件下保护酶和提高酶效率的能力。该项目吸引了来自历史上代表性不足的科学群体(IHUGS)的学生。对高中生的推广和对IHUGS参与者的研究培训是该项目的一部分。这个合成生物学项目结合了两个自然系统和一个人工系统来构建和测试一套独特的移动酶隔离平台(MESP)。由此设计的体系提高了在低pH和高温条件下的酶活性。该嵌合平台由来自Sulfolobus Group II伴侣蛋白复合体(热休克蛋白复合体)的精选亚基和来自纤维素分解厌氧细菌热梭菌的成分组成。根据用途，可以结合到平台上的酶可以是一种或多种酶类。这项研究提出了关于合成系统的结构和功能的基本假设，并检查了多酶系统的组装和功能的原则。该项目使用了微生物学、分子生物学、蛋白质生物化学和纳米技术的方法。这一平台技术可用于生物燃料生产、生物修复和许多其他应用。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

This project is co-funded by the Systems and Synthetic Biology cluster in the Division of Molecular and Cellular Biosciences and the Human Resources cluster in the Division of Biological Infrastructure.The need for alternative transportation fuels that can supplement or replace fossil-based fuels (e.g., gasoline and diesel) is an ongoing global concern. The same chemical conversion technologies that are used to produce bioethanol and biodiesel are being used to produce biofuel. This process involves the use of harsh chemicals (e.g., acids) that create a hazardous materials handling and disposal problem for producers. Enzyme-based conversion of feedstock to biofuel presents fewer hazards and is, arguably, more environment-friendly. Unfortunately, microbial or abiotic enzyme-mediated reactions are typically not as efficient as chemical catalysis. This project is focused on designing and testing an experimental enzyme sequestration platform system that enhances enzymatic efficiency under conditions of high temperature and extremes of pH. Such conditions are common in industrial reaction processes. The project has its foundation in fundamental biochemistry research in which mobile enzyme sequestration platforms are modified and examined for effectiveness. The platforms are investigated to characterize their ability to protect enzymes and enhance enzymatic efficiency under harsh reaction conditions. The project engages students who are part of the Individuals from Historically Underrepresented Groups in Science (IHUGS). Outreach to high school students and research training for IHUGS participants are activities that form part of the project.This project in synthetic biology combines two natural systems and one artificial system in constructing and testing a set of unique mobile enzyme sequestration platforms (mESPs). The resulting designed system enhances enzymatic activity under conditions of low pH and high temperature. The chimeric platform consists of select subunits from the Sulfolobus group II chaperonin complex (heat shock protein complex) and components from the cellulolytic anaerobic bacterium Clostridium thermocellum. Enzymes that can bind to the platform may be of one or more enzyme classes depending on use. This research project addresses fundamental hypotheses on the synthetic system's structure and function and examines the principles for the assembly and function in multi-enzyme systems. The project uses methods of microbiology, molecular biology, protein biochemistry and nanotechnology. This platform technology may be used in biofuel production, bioremediation, and a host of other applications.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.