CAREER: Designing a Synthetic Nucleolus for Cell Free Biocatalysis

非技术摘要：这项研究将开发含有酶的新生物材料。酶是一种功能性蛋白质，在自然界中充当化学反应的催化剂。与目前以石油为基础的工业方法相比，它们有可能以高效、选择性和环保的方式制造燃料、药品和精细化学品。酶的全部力量尚未在化学制造中释放出来，因为在自然界中，酶与其他酶在复杂的网络中工作，在温和的条件下，这与工业过程不相容。这项提议研究了如何使用新的生物材料来创建像自然界中的那些有用的多酶系统的基础科学。此外，K-12、本科生和研究生的生物材料教育通过发展双语(英语/西班牙语)经验和内容被整合到这项计划中，旨在增加拉美裔人对科学、技术、工程和数学事业的参与。技术摘要：与目前占主导地位的方法：基于石油的合成化学相比，酶可以以高效、选择性和环保的方式合成化学产品。然而，由于将细胞外的多酶途径整合到化学制造中的方法不足，生物催化的全部力量尚未释放。自然界有通过时空调节来控制多酶反应的有效策略。例如，像核仁这样的细胞内结构是由多个相组成的液滴，在每个相中限制不同的酶来组装核糖体。核仁能够通过隔离必要的成分和排除其他成分在细胞复杂的内部发挥作用。此外，在细胞周期的不同阶段需要时，核仁可以动态地组装和拆解。如果核仁的结构能够被模拟并用于在细胞环境外进行多酶反应，这将通过提供更高的产率和减少成本和浪费来改变化学制造。这项建议研究重建核仁结构和模拟核仁功能所需的基础生物材料科学。一个基于多肽的复杂凝聚体的库，形成类似于核仁的液滴，将被设计成具有不同的界面张力和包埋选择性。该库将用于评估实现稳定的多相凝聚所需的条件，并测量添加剂在相内和相间的扩散。最后，将使用酶串联反应来测试不同合成核仁的功能，并将其与相的材料性质相关联。在整个提案中，“生物材料相分离”的科学将被用来增加西班牙裔学生在STEM职业生涯中的参与度。这一努力的关键是为10-13岁的儿童开发一个体验式和双语(英语/西班牙语)暑期课程，旨在改善家庭中的科学交流，这是选择STEM职业的关键指标。该计划与向青少年介绍“生物材料相分离”科学，并向本科生和研究生介绍课程作业和实验室经验的双语社交媒体活动相结合。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 Abstract: This research will develop new biomaterials incorporating enzymes. Enzymes are functional proteins that act as a catalyst for chemical reactions in nature. They have the potential to make fuels, pharmaceuticals, and fine chemicals in efficient, selective, and ecofriendly ways compared to current industrial methods based on petroleum. The full power of enzymes has yet to be unleashed on chemical manufacturing because in nature enzymes work with other enzymes in complex networks, under mild conditions, that are incompatible with industrial processes. This proposal investigates the fundamental science of how new biomaterials can be used to create useful multi-enzyme systems like those in nature. In addition, biomaterials education of K-12, undergraduate, and graduate students is integrated throughout this proposal through the development of bilingual (English/Spanish) experiences and content aimed at increasing Hispanic participation in science, technology, engineering, and math careers.Technical Abstract: Enzymes can synthesize chemical products in efficient, selective, and ecofriendly ways compared to the current dominant method: petroleum-based synthetic chemistry. However, the full power of biocatalysis has yet to be unleashed due to inadequate methods of integrating multienzyme pathways outside of cells into chemical manufacturing. Nature has efficient strategies to control multienzyme reactions using spatiotemporal regulation. For instance, intracellular structures like the nucleolus, which are liquid droplets consisting of multiple phases, confine different enzymes in each phase to assemble the ribosome. The nucleolus is able to function in the complex interior of the cell by sequestering the necessary components and excluding others. In addition, the nucleolus can then dynamically assemble and disassemble when needed in different stages of the cell cycle. If the structure of the nucleolus can be mimicked and used to perform multienzyme reactions outside of a cellular environment, this would transform chemical manufacturing by providing higher yields and reducing cost and waste. This proposal investigates the fundamental biomaterials science required to recreate the structure and mimic the function of the nucleolus. A library of peptide based complex coacervates, that form liquid droplets similar to the nucleolus, will be designed with varying interfacial tensions and encapsulation selectivity. This library will be used to evaluate the conditions needed to achieve stable multiphase coacervates and measure diffusion of additives within and between the phases. Finally, the function of different synthetic nucleoli will be tested using an enzyme cascade reaction and correlated to the material properties of the phases. Throughout this proposal the science of "biomaterials phase separation" will be used to increase the participation of Hispanic students in STEM careers. Key to this effort is the development of an experiential and bilingual (English/Spanish) summer program for ages 10-13 geared to improve science communication at home, a key indicator of choosing STEM careers. This program is paired with a bilingual social media campaign introducing "biomaterials phase separation" science to teenagers and coursework and laboratory experiences for undergraduates and graduate students.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.