CAREER: Surfactant Proteins that Stabilize Biomolecular Condensates: From Biophysics to Biomaterials for Biomanufacturing

非技术描述：生物聚合物是由生物体产生的大而复杂的分子。几千年来，人类一直使用生物聚合物为基础的材料来满足他们的日常需求：木材建造房屋和羊毛编织衣服是两个众所周知的例子。在现代，我们如何继续从自然中汲取灵感，开发新型生物高分子材料，以满足人类最迫切的需求？活细胞由成千上万种不同的生物聚合物组成，这些聚合物可以自我组织，从而产生生物结构和活性。该项目的目标是了解生物聚合物如何在细胞中自组装，然后将这些见解应用于新生物材料的开发。这个职业项目将专注于蛋白质，这是一大类重要的生物聚合物。首席研究员和他的团队将研究生物分子凝聚物，这是一种在细胞内自发聚集的富含蛋白质的球形液滴，以及表面活性剂蛋白质，表面活性剂蛋白质覆盖在这些生物分子凝聚物的表面，赋予其稳定性和功能。研究人员将首先识别和设计表面活性剂蛋白质，然后确定表面活性剂与生物分子凝聚物相互作用和影响的物理和化学原理。基于这些研究，研究人员将开发生物材料来解决药物合成中的一个重要问题。许多蛋白质是酶，其功能是加速生化反应。酶催化反应是传统化学催化的强大替代品，可以实现负担得起的“绿色”制药。然而，酶的纯化和配方是阻碍生物催化领域发展的主要挑战。研究人员建议通过开发基于酶活性、表面活性剂涂层的生物分子凝聚物的新型仿生材料来解决这些挑战。因此，本研究旨在推进对细胞如何构建材料的基本理解，然后将利用这一基本理解来设计用于药物生物催化的生物材料。该研究项目将与“生化工程师伦理”教育项目相结合。该教育计划的目标是培养学生考虑如何通过他们的职业生涯来解决主要的道德挑战，包括环境可持续性和公平的全球获得负担得起的药物。技术描述：细胞划分其内部以协调其在空间和时间上的生化过程。生物分子凝聚物是通过蛋白质和其他生物聚合物相分离形成的细胞隔室。因此，细胞质使人联想到乳剂，由一相的液滴分散在另一相中。在消费者应用中使用的乳剂需要表面活性剂来赋予稳定性和功能，因此化学表面活性剂被广泛使用和研究。然而，研究人员很少关注细胞内蛋白质表面活性剂及其对细胞乳状组织的贡献。为了弥合这一知识差距，该职业奖旨在阐明“表面组”-在生物分子凝聚物表面自组装的两亲性蛋白质，稳定凝聚物并调节其生物物理性质和功能。研究人员建议确定控制这种细胞内表面活性的物理化学原理，量化这种表面活性蛋白对生物分子凝聚聚结的影响，并进行生物信息学调查以绘制表面组的全部范围。这项基础研究为解决生物制造中的一个棘手问题提供了灵感。在现代制药工业中，人们对用体外酶催化反应取代传统的催化剂材料以更可持续、更安全、更经济地合成药用化合物有着浓厚的兴趣。然而，酶的纯化和稳定性是阻碍药物生物催化的关键挑战。受细胞利用生物分子凝聚物划分和调节酶促反应的启发，研究人员提出开发药物生物合成的使能技术。具体来说，研究人员将设计由交联、表面活性剂包裹的蛋白质凝聚物组成的固定化酶材料。其基本原理是，这些生物材料将同时允许通过相分离进行简便、无色谱的酶纯化，并将在冷凝物表面最佳地显示酶，以获得最大的酶活性。本研究将阐明“表面组”科学的基本原理，开发用于药物生物制造的表面活性固定化酶生物材料。本计画的教育部分旨在培养学生在生化工程中以伦理为基础的决策。这个伦理项目将包括一个针对缺乏服务的本科生的暑期研究项目，重点是生物材料和可持续性；工科研究生的职业道德培养大四本科生课程模块，以生化工程伦理案例研究为特色。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

Non-Technical Description:Biopolymers are large, complex molecules produced by living organisms. For millennia, humans have used biopolymer-based materials to meet their everyday needs: wood to build homes and wool to weave clothes are two well-known examples. In the modern era, how can we continue to derive inspiration from nature to develop novel biopolymeric materials for humanity’s most pressing needs? Living cells are comprised of many thousands of different biopolymers that self-organize, giving rise to biological structure and activity. The goal of this project is to learn how biopolymers self-assemble in cells and then apply those insights for the development of new biomaterials. This CAREER project will focus on proteins, a large and important class of biopolymers. The principal investigator and his team will study biomolecular condensates, which are protein-rich spherical droplets that spontaneously assemble inside cells, and surfactant proteins that coat the surface of these biomolecular condensates and impart stability and function. The investigators will first identify and engineer surfactant proteins, then determine the physical and chemical principles by which surfactants interact with and influence biomolecular condensates. Based on these studies, the investigators will develop biomaterials to address an important problem in pharmaceutical synthesis. Many proteins are enzymes, whose function is to accelerate biochemical reactions. Enzyme-catalyzed reactions are a powerful alternative to traditional chemical catalysis to enable affordable, “green” pharmaceutical manufacturing. However, purification and formulation of enzymes are major challenges that impede advancement of the biocatalysis field. The investigators propose to address these challenges by developing new bio-inspired materials based on enzymatically active, surfactant-coated biomolecular condensates. Thus, this research seeks to advance fundamental understanding of how cells build materials, then will leverage this fundamental understanding to engineer biomaterials for pharmaceutical biocatalysis. This research program will be coupled with an education program, “Ethics for Biochemical Engineers.” The goal of this education program is to train students to consider how through their careers they can address major ethical challenges, including environmental sustainability and equitable global access to affordable medications.Technical Description:Cells compartmentalize their interiors to orchestrate their biochemical processes in space and time. Biomolecular condensates are cellular compartments formed via phase separation of proteins and other biopolymers. The cytoplasm is therefore reminiscent of an emulsion, comprising droplets of one phase dispersed in another phase. Emulsions used in consumer applications require surfactants to impart stability and function, so chemical surfactants are widely used and studied. However, researchers have devoted scant attention to intracellular protein surfactants and their contribution to the emulsion-like organization of the cell. To bridge that knowledge gap, this CAREER award aims to elucidate the “surfactome” – amphiphilic proteins that self-assemble at the surface of biomolecular condensates, stabilizing the condensates and regulating their biophysical properties and functions. The investigators propose to identify the physicochemical principles governing such intracellular surfactancy, quantify the effect of such surfactant proteins on biomolecular condensate coalescence, and conduct a bioinformatics survey to map the full scope of the surfactome. This fundamental research inspires a solution to a vexing problem in biomanufacturing. In the modern pharmaceutical industry, there is keen interest in replacing traditional catalyst materials with in vitro enzyme-catalyzed reactions to synthesize medicinal compounds more sustainably, safely, and affordably. However, enzyme purification and stability are critical challenges that have hindered pharmaceutical biocatalysis. Inspired by cells’ use of biomolecular condensates to compartmentalize and regulate enzymatic reactions, the investigators propose to develop enabling technologies for pharmaceutical biosynthesis. Specifically, the investigators will engineer immobilized enzyme materials comprised of crosslinked, surfactant-coated protein condensates. The rationale is that these biomaterials will simultaneously allow facile, chromatography-free enzyme purification via phase separation, and will optimally display enzymes at the condensate surface for maximal enzyme activity. Together, this research will elucidate basic principles of “surfactome” science and develop surface-active immobilized enzyme biomaterials for pharmaceutical biomanufacturing. The education component of this project aims to train students in ethics-based decision making in biochemical engineering. This ethics program will include a summer research program for underserved undergraduates, focusing on biomaterials and sustainability; ethics training for engineering graduate students; and a course module for senior undergraduates, featuring case studies in biochemical engineering ethics.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.