CAREER: Intein-Mediated Protein Polymer Hydrogels as General Scaffolds for Immobilization of Bioactive Proteins

这项由生物技术、生化和生物质工程项目授予的NSF CAREER奖支持一种新型蛋白质聚合物水凝胶的工程设计，作为固定化酶和生物活性蛋白的通用支架。酶是多功能催化剂，在现代生物技术中发挥着越来越重要的作用。然而，酶在溶液中的长期稳定性差，回收和再循环困难，极大地阻碍了它们在生物转化中的应用。将酶固定在固体载体上可以显著提高酶的稳定性，便于酶的循环利用和回收。所提出的蛋白质水凝胶利用内部介导的蛋白质剪接/连接来形成一个相互连接的蛋白质聚合物网络。该项目的成功完成将为蛋白质水凝胶的合成提供一种通用的方法，这种方法可以密集有效地结合多种生物活性蛋白质，并具有高度控制的分子结构。该项目将产生更广泛的影响：(1)通过扩大现有的蛋白质固定化工具；(2)通过来自不同背景的年轻和有抱负的科学家的参与。这项工作的技术目标是探索蛋白质固定化的新领域。从这些研究中获得的见解将使几个采用生物催化工艺的行业受益，提高我们为生物转化和酶燃料电池组装高效酶途径的能力，并促进新的生物活性蛋白水凝胶支架的创建，用于组织工程和药物输送应用。提高公众对生物技术/水凝胶认识的教育目标将通过以下方式实现：(1)建立一个以水凝胶和蛋白质自组装为重点的互动网络空间和YouTube视频模块；(2)设计以水凝胶为重点的动手实验模块，在已建立的本科课程和TAMU外展活动中展示，包括化学周和物理日；(3)通过E3教师研究体验（RET）计划主持和指导中学科学和数学教师。(4)在一所以少数民族为主的高中，通过课堂讨论对拟议研究中探讨的主题进行教育；(5)与当地记者合作，以外行可以理解和欣赏的语言传播研究成果；(6)在实验室接待/指导本科生，目标是少数民族学生。

CBET 1150478/ ChenThis NSF CAREER award by the Biotechnology, Biochemical and Biomass Engineering program supports the engineering of a novel protein polymer hydrogel as a general scaffold for the immobilization of enzymes and bioactive proteins. Enzymes are versatile catalysts and are playing an ever-more prominent role in modern biotechnology. However, poor long-term stability and difficulties in recovery and recycling of enzymes in solution have greatly hampered their usefulness in biotransformation. Immobilization of enzymes on solid supports can significantly enhance their stability and enable convenient recycling and recovery. The proposed protein hydrogel takes advantage of intein-mediated protein splicing/ligation to form an inter-connected network of protein polymers. Successful completion of this project will provide a general method for the synthesis of protein hydrogels that densely and efficiently incorporate multiple bioactive proteins with a highly controlled molecular architecture.This program will achieve broader impacts (1) by expanding the current toolkit for protein immobilization and (2) through participation of young and aspiring scientists from diverse backgrounds. The technical goal of this work explores new frontiers in protein immobilization. Insights from these studies will benefit several industries employing biocatalytic processes, advance our ability to assemble efficient enzymatic pathways for biotransformation and enzymatic fuel cells, and facilitate the creation of new bioactive protein hydrogel scaffolds for tissue engineering and drug delivery applications. The educational goal of increasing public awareness in biotechnology/hydrogels will be achieved through: (1) establishing an interactive web-space and YouTube video modules focused on hydrogels and protein self-assembly, (2) designing hands-on hydrogel-focused experimental modules for showcasing in an established undergraduate course and at TAMU outreach outlets including Chemistry Week and Physics Day, (3) hosting and mentoring secondary science and math teachers through the E3 Research Experience for Teachers (RET) Program, (4) educating juniors at a predominantly minority high school through class discussions on topics explored in the proposed research, (5) disseminating research results in a language understandable and appreciable by laypersons in collaboration with local journalists, and (6) hosting/mentoring undergraduate students in the lab, targeting minority students.