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

合成生物学：结构改变对移动酶封存平台 (mESP) 功能的影响

基本信息

批准号：
1818346
负责人：
Ruben Ceballos
金额：
$ 80万
依托单位：
University of Arkansas
依托单位国家：
美国
项目类别：
Standard Grant
财政年份：
2018
资助国家：
美国
起止时间：
2018-08-01 至 2023-07-31
项目状态：
已结题

来源：
https://www.nsf.gov/awardsearch/showAward?AWD_ID=1818346&HistoricalAwards=false
关键词：
Synthetic Biology Impacts structural alterations

项目摘要

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.

该项目由系统和合成生物学群集共同资助分子和细胞生物科学的划分以及人力资源集群在生物基础设施的划分中。需要补充或取代基于化石的燃料的替代运输燃料（例如，汽油和柴油）是一个启发的全球关注。用于生产生物乙醇和生物柴油的相同化学转化技术被用于生产生物燃料。此过程涉及使用刺激性化学物质（例如酸），这些化学物质会为生产者带来危险的材料处理和处置问题。基于酶的原料转换为生物燃料的危险更少，并且可以说是环境友好的。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有效。该项目的重点是设计和测试实验酶隔离平台系统，该系统在高温和极端pH的条件下提高了酶促效率。这种情况在工业反应过程中很常见。该项目在基本生物化学研究中奠定了基础，在该研究中，对移动酶隔离平台进行了修改并检查有效性。研究了平台以表征其保护酶的能力，并在恶劣的反应条件下提高酶促效率。该项目吸引了来自历史上代表性不足的科学（IHUGS）的学生的一部分。向高中生和IHUGS参与者的研究培训是构成该项目的一部分的活动。该合成生物学的项目结合了两个自然系统和一个人工系统，用于构建和测试一组独特的移动酶隔离平台（MESPS）。所得设计的系统在低pH和高温条件下增强了酶活性。嵌合平台由来自磺胺糖II组伴侣蛋白复合物（热休克蛋白复合物）的精选亚基和来自纤维溶解厌氧菌细菌热环状的成分组成。可以与平台结合的酶可能是一个或多个酶类，具体取决于使用。该研究项目介绍了关于合成系统的结构和功能的基本假设，并研究了多酶系统中组装和功能的原理。该项目使用微生物学，分子生物学，蛋白质生物化学和纳米技术的方法。该平台技术可用于生物燃料生产，生物修复和许多其他应用程序中。该奖项反映了NSF的法定任务，并被认为是值得通过基金会的知识分子优点和更广泛的影响评估标准通过评估来支持的。