CAREER: Investigating Non-heme Iron Enzyme Catalyzed Nitrile Formation Mechanisms in Etoposide and Other Natural Product Biosynthetic Pathways

酶是生物催化剂，可以加快细胞内化学反应的速度。重新定向和重新利用酶来生产理想的新型化学产品是一项挑战，但可能具有重大的工业回报。有了这个奖项，化学部的生命过程化学项目正在资助来自北卡罗来纳州州立大学的Wei-chen Chang博士重新利用特定的含铁酶来加速这些蛋白质通常不执行的各种化学反应。最近的计算和机械学进展指出了使用这些酶来“生物合成”工业感兴趣的化合物的机会，并了解如何控制这些反应的基本原理。Chang博士的研究使用了一种“底物诱饵”，其中标准起始化合物被一种新的反应物取代，以改变酶催化的化学类型。在这个项目中，含铁的酶被重新编程以改变它们的功能。 他们过去分解分子，现在添加化学基团来创造新的化合物。研究计划与教育活动相结合，以激励下一代学生在化学和生物学之间的界面上进行跨学科研究。“科学快车”外展活动利用一个简单的生化反应，将普通的糖转化为蓝色染料（“蓝色牛仔裤的蓝色基因”动手体验），向学生们展示化学生物学如何影响他们周围的世界。这些活动吸引了来自北卡罗来纳州农村地区不同文化、种族和社会经济背景的代表性不足的K-12学生。该研究项目旨在开发一种底物诱饵方法，以重定向单核非血红素铁和2-酮戊二酸依赖性（Fe/2 OG）催化的反应，同时保持其催化效率。Chang博士的目标是探索利用这种方法来生物合成工业上重要分子的潜力。研究目的是将这些酶的反应产物从羟基化转向腈基安装。一个互补的目标是阐明，通过各种化学探测和动力学方法，Fe/2 OG羟化酶催化这种非天然反应的机制。张博士的教育目标包括开发酶动力学本科课程，通过积极的学习方法整合化学和生物化学概念。此外，他的团队还开发了一种易于理解的酶催化反应（“蓝色牛仔裤的蓝色基因”），作为科学快车推广计划的一部分。该项目是与当地教师合作设立的，旨在向中学生介绍令人兴奋的化学生物学研究。该奖项反映了NSF的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

Enzymes are biological catalysts that increase the speed of chemical reactions in the cell. Redirecting and repurposing enzymes to produce desirable novel chemical products is a challenge but could have significant industrial rewards. With this award, the Chemistry of Life Processes Program in the Chemistry Division is funding Dr. Wei-chen Chang from North Carolina State University to repurpose specific iron-containing enzymes to speed-up a variety of chemical reactions that these proteins do not typically perform. Recent computational and mechanistic advances point to opportunities to use these enzymes to "bio-synthesize" compounds of industrial interest, and to understand the fundamental principles of how to control such reactions. Dr. Chang's studies use a "substrate decoy", in which the standard starting compound is replaced by a new reactant to change the type of chemistry catalyzed by the enzyme. For this project, the iron-containing enzymes are reprogramed to alter their functions. Where they used to break down molecules, they now add chemical groups to create new compounds. The research plan is integrated with educational activities to inspire the next generation of students to pursue interdisciplinary research at the interface between chemistry and biology. The "science express" outreach activity uses a simple biochemical reaction that turns ordinary sugar into a blue dye (the "blue genes for blue jeans" hands-on experience) to show students how chemical biology impacts the world around them. These activities engage underrepresented K-12 students from diverse cultural, racial, and socioeconomic backgrounds in rural areas of North Carolina. This research project aims to develop a substrate decoy approach to redirect the reactions catalyzed by mononuclear non-heme iron and 2-oxoglutarate dependent (Fe/2OG) while maintaining their catalytic efficiencies. Dr. Chang's goal is to explore the potential of utilizing this approach to biosynthesize industrially important molecules. The research objective is to redirect the reaction outcome of these enzymes from hydroxylation to nitrile group installation. A complementary objective is to elucidate, through various chemical probing and kinetic approaches, the mechanism by which Fe/2OG hydroxylases catalyze this non-native reaction. Dr. Chang's educational objectives include developing an undergraduate course in enzyme kinetics that integrates chemical and biochemical concepts through active learning methods. In addition, his group has developed an easy-to-understand enzyme catalyzed reaction ("blue genes for blue jeans") as part of the science express outreach program. This program, established in collaboration with local teachers, introduces middle school students to exciting research in chemical biology.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.