CAREER: Room Temperature Stabilization of Cellular Factories by Confinement: A Thermodynamic Approach

摘要：CBET-0644784， U. Minnesota, Dr. A. AksanCAREER：通过限制室温稳定细胞工厂：一种热力学方法。该职业项目的目标是开发一个高度集成的研究和教学计划，重点关注生物系统（蛋白质，生物膜和细胞工厂）的热力学稳定。细胞工厂是细菌、植物或动物的细胞，它们经过基因工程改造，可以产生特定的蛋白质，或进行复杂的化学反应。在室温下通过封闭保存生物系统（通过包埋在糖玻璃中，或封装在多孔基质中）正在成为一种可行的和经济的替代传统技术，需要低温处理或储存。某些酶和蛋白质在二氧化硅基质约束下的长期室温稳定已经成为可能。然而，对于更敏感的蛋白质或细胞工厂来说，同样的成功程度是无法重复的。失败的原因尚不清楚。这就是PI开发他的程序的动力。智力优势：本项目的研究目标是建立约束生物系统室温稳定的热力学。这将通过追求以下具体目标来实现：1)建立水活度变化对生物系统损害的机制、动力学和热力学；2)建立约束作用下生物系统室温稳定的机制和热力学；3)建立约束作用下室温稳定的热力学模型。该提案的综合教学目标是开发一种新颖的、基于技术的教育工具（e - value计划），其重点是通过增加本科生和高中生（以及教师）在科学和工程教育和研究活动中的曝光和参与来增加外联和指导。本计划的目标是：生物隔离系统被用于生物医学（作为糖尿病患者的胰岛素分泌植入物）、生物技术（重组蛋白质和酶的生产）、生物修复（清理有毒废物和污染物）、绿色化学、替代能源（利用细菌发电和制氢）和生物传感器（“金丝雀细胞”）。可以检测污染物、病毒制剂或有毒化学物质的存在。通过PI与明尼苏达大学机械工程系、微生物学系、生物技术研究所和当地工业的合作，这里提出的研究活动将实验与理论建模从多学科的角度结合起来。这为通过该项目就业的本科生和研究生创造了一个独特的研究和教学环境。教学程序使用了一种非常普遍的技术工具（一台可以上网的个人电脑）。这使得PI更容易接触到代表性不足的群体和地理上处于不利地位的机构。作为教学计划的一部分，还将开发一门本科/研究生水平的课程，重点是热力学原理在自然现象中的应用。

ABSTRACT: CBET-0644784 , U. Minnesota, Dr. A. AksanCAREER: Room Temperature Stabilization of Cellular Factories by Confinement: A Thermodynamic Approach The goal of this CAREER project is to develop a highly integrated research and teaching program focused on the thermodynamics of stabilization of biological systems (proteins, biomembranes, and cellular factories). Cellular factories are bacteria, plant, or animal cells that are genetically engineered to produce specific proteins, or to perform complex chemical reactions.). Preservation of biological systems at room temperature by confinement (by embedding in sugar glasses, or encapsulation in porous matrices) is emerging as a viable and economical alternative to traditional techniques that require cryogenic temperatures for processing or storage. Long-term room temperature stabilization of certain enzymes and proteins in silica matrix confinement has been possible. However, the same level of success could not be repeated for more sensitive proteins, or cellular factories. The reason for failure is not known. This is the driving force for the PI to develop his program. Intellectual Merit: The research objective of this project is to establish the thermodynamics of room temperature stabilization of biological systems by confinement. This will be achieved by pursuing the following specific aims: 1) Establish the mechanism(s), kinetics and thermodynamics of damage to biological systems due to changes in water activity, 2) Establish the mechanism(s), and thermodynamics of room temperature stabilization of biological systems by confinement, 3) Develop a thermodynamic model for room temperature stabilization by confinement. The integrated teaching objective of the proposal is to develop a novel, technology-based educational tool (the E-valuate Program), which is focused on increasing outreach and mentoring by increasing the exposure and involvement of undergraduate and high school students (as well as teachers) in scientific and engineering education and research activities. The objectives of this program are:Broader Impacts: The biological confinement systems are used in biomedicine (as insulin secreting implants for diabetic patients), biotechnology (recombinant protein, and enzyme production), bioremediation (to clean up toxic waste, and pollutants), green chemistry, alternative energy generation (electricity and hydrogen production by bacteria), and as biosensors ("canary cells," which can detect the presence of pollutants, viral agents, or toxic chemicalsThe research activities proposed here combine experimentation with theoretical modeling from a multidisciplinary perspective through the collaborations the PI has fostered with the Departments of Mechanical Engineering, and Microbiology, the Biotechnology Institute at the University of Minnesota and the local industry. This creates a unique research and teaching environment for undergraduate and graduate students employed through this project. The teaching program utilizes a very commonly available technological tool (a PC with an internet connection). This makes it easier for the PI to reach the underrepresented groups and geographically disadvantaged institutions. As part of the teaching plan, a combined undergraduate/graduate level course focusing on the applications of thermodynamic principles to natural phenomena will also be developed.