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

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 immobilization of enzymes and bioactive proteins.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，差的长期稳定性和在溶液中的酶的回收和再循环的困难极大地阻碍了它们在生物转化中的有用性。将酶固定在固体载体上可以显著提高其稳定性，并且能够方便地再循环和回收。所提出的蛋白质水凝胶利用内含肽介导的蛋白质剪接/连接形成蛋白质聚合物的互连网络。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，该方法可以密集有效地将多种生物活性蛋白质与高度受控的分子结构结合在一起。该项目将通过（1）扩展现有的蛋白质固定化工具包以及（2）通过来自不同背景的年轻和有抱负的科学家的参与来实现更广泛的影响。这项工作的技术目标是探索蛋白质固定化的新前沿。这些研究的见解将有利于采用生物催化过程的几个行业，提高我们组装生物转化和酶促燃料电池的有效酶促途径的能力，并促进为组织工程和药物递送应用创造新的生物活性蛋白质水凝胶支架。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）建立一个互动的网络空间和YouTube视频模块，重点是水凝胶和蛋白质自组装，（2）设计动手水凝胶为重点的实验模块，展示在一个既定的本科课程和TAMU外展网点，包括化学周和物理日，（3）通过E3教师研究经验（RET）计划主持和指导中学科学和数学教师，（4）通过课堂讨论在拟议的研究中探索的主题，在一所以少数民族为主的高中教育三年级学生，（5）与当地记者合作，以外行人可以理解和理解的语言传播研究成果，以及（6）在实验室接待/指导本科生，针对少数民族学生。

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.