Engineering biocatalytic nanoreactor

区隔化是细胞功能的一个重要组成部分，它使多种生化反应在细胞的特定位置和特定时间发生。在实验室和工业中，用一种或多种生物和化学催化剂再现这种性能，为绿色和可持续化学过程的未来发展提供了巨大的机会。该项目从自然界中汲取灵感，探索细菌“胶囊”作为有序固定催化剂的容器。胶囊是纳米级的空心球体，带有自发组装的蛋白质外壳。蛋白质工程策略将探索定制这些纳米结构的特定反应序列感兴趣。目标是提高原料通过壳的选择性通道，并用特定的反应催化剂可重复地“装饰”壳的内部。综合起来，这些努力将产生工程化的纳米级反应堆容器，可以设计成执行各种工业上有趣的反应序列。还将有STEM职业发展努力，涉及从K-12到大学年龄的学生。其中包括小学的科学日，当地高中生的蛋白质建模比赛，本科生的研究经历，以及每年亚特兰大科学节的蛋白质设计和3D打印活动。该项目将探索（生物）催化剂的催化性能对封装和控制表面固定化的影响，以蛋白质为基础的纳米室封装。从随机诱变和增量截断到蛋白质设计的蛋白质工程策略将被用于重塑纳米室，以增强底物和产品的渗透性，并通过在包被表面的人工亲和位点创造功能多样化的新机会。除了单颗粒之外，将纳米室组装成高密度生物催化剂系统的多维纳米反应器组件将被探索。结合使用生物物理方法，以及生物催化和生物感觉系统，将评估这些工程努力对纳米室本身结构和相关催化剂功能的影响。通过将设计参数集成到该系统中，该系统具有高对映体选择性和降低分离成本的潜力，该技术将对治疗药物开发产生重大影响。

Compartmentalization is a critical component of cellular function, enabling a multitude of biochemical reactions to occur at specific location in the cell at specific times. Reproducing such performance with one or more biological and chemical catalysts in the lab and in industry offers tremendous opportunities for future developments of green and sustainable chemical processes. The proposed project draws inspiration from nature by exploring bacterial "capsules" as containers for the orderly immobilization of catalysts. The capsules are nanometer-scale, hollow spheres with protein shells that assemble spontaneously. Protein engineering strategies will be explored to tailor these nanostructures to the specific reaction sequences of interest. The goals are to improve the selective passage of raw materials through the shell, and to reproducibly "decorate" the interior side of the shell with specific reaction catalysts. In combination, these efforts will produce engineered nano-scale reactor vessels that can be designed to perform a variety of industrially interesting reaction sequences. There will also be STEM career development efforts that will involve students from K-12 through college age. These include Science Days at an elementary school, protein modeling competitions for local high school students, undergraduate research experiences, and protein design and 3D printing activities at the Annual Atlanta Science Festival.The project will explore the impact of catalytic performance of (bio) catalysts upon encapsulation in and controlled surface-immobilization on protein-based encapsulin nanocompartments. Protein engineering strategies ranging from random mutagenesis and incremental truncation to protein design will be employed to remodel nanocompartments for enhanced substrate and product permeability and to create new opportunities for functional diversification via artificial affinity sites on the encapsulin surfaces. Beyond single particles, the assembly of nanocompartments into multi-dimensional nanoreactor assemblies for high-density biocatalyst systems will be explored. Using a combination of biophysical methods, as well as biocatalytic and biosensory systems, the impact of these engineering efforts on the structure of nanocompartments themselves and the function of the associated catalysts will be assessed. By integrating design parameters into this system that offer the potential for high enantioselectivity and reduced separation costs, this technology would have significant implications for therapeutic drug development.