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

非技术描述：生物聚合物是由活的有机体产生的大而复杂的分子。几千年来，人类一直使用基于生物聚合物的材料来满足他们的日常需求：建造房屋的木材和编织衣服的羊毛是两个众所周知的例子。在现代，我们如何继续从大自然中获得灵感，为人类最紧迫的需求开发新的生物聚合物材料？活细胞由数千种不同的生物聚合物组成，这些聚合物自组织起来，产生生物结构和活动。这个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，这是在细胞内自发组装的富含蛋白质的球形液滴，以及覆盖在这些生物分子凝聚物表面并赋予稳定性和功能的表面活性蛋白质。研究人员将首先识别和设计表面活性蛋白质，然后确定表面活性物质与生物分子缩合物相互作用和影响的物理和化学原理。在这些研究的基础上，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一种强有力的替代方案，可以实现负担得起的“绿色”制药生产。然而，酶的纯化和制备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缩合物的新的生物灵感材料来应对这些挑战。因此，这项研究旨在促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。这项研究计划将与“生化工程师伦理”教育计划相结合。这个教育项目的目标是训练学生思考如何在他们的职业生涯中解决主要的伦理挑战，包括环境的可持续性和全球公平地获得负担得起的药物。技术描述：细胞内部分隔，以协调他们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的相分离形成的细胞室。因此，细胞质使人联想到乳状液，由分散在另一相中的一相液滴组成。消费应用中使用的乳状液需要表面活性剂提供稳定性和功能，因此化学表面活性剂得到了广泛的应用和研究。然而，研究人员对细胞内蛋白表面活性物质及其对细胞乳状液样组织的贡献关注较少。为了弥合这一知识鸿沟，这一职业奖旨在阐明“表面蛋白质组”--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。研究人员建议确定支配这种细胞内表面作用的物理化学原理，量化这种表面活性蛋白对生物分子凝析油结合的影响，并进行生物信息学调查，以绘制表面酶组的全部范围。这项基础性研究启发了生物制造中一个棘手问题的解决方案。在现代制药工业中，用体外酶催化反应取代传统的催化剂材料，以更可持续、安全和经济的方式合成药物化合物是人们非常感兴趣的。然而，酶的纯化和稳定性是阻碍药物生物催化的关键挑战。受细胞使用生物分子缩合物来区分和调节酶反应的启发，研究人员提议开发使能技术用于药物生物合成。具体地说，研究人员将设计由交联的、表面活性物质包裹的蛋白质凝聚体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单、无层析的酶纯化，并将最佳地在冷凝液表面展示酶以获得最大的酶活性。这项研究将阐明“表面组学”的基本原理，并开发用于制药生物制造的表面活性固定化酶生物材料。该项目的教育部分旨在培训学生在生化工程中进行基于伦理的决策。这一伦理学计划将包括面向本科生的暑期研究计划，重点是生物材料和可持续发展；为工程学研究生提供伦理学培训；以及为大四本科生开设课程模块，以生化工程伦理学的案例研究为特色。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。

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.