Synthetic Biology Approaches to Selective Amino Acid Modification and Incorporation

选择性氨基酸修饰和掺入的合成生物学方法

• 批准号: 2204014
• 负责人: Michelle Chang
• 金额: $ 52.8万
• 依托单位: University of California-Berkeley
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2025-08-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2204014&HistoricalAwards=false
• 关键词: Synthetic; Biology; Approaches; Selective; Amino

With the support of the Chemistry of Life Processes Program in the Chemistry Division, Professor Michelle Chang from the University of California, Berkeley will develop methods for the incorporation of non-canonical amino acids into proteins. Although proteins carry out a broad range of functions in the cell, they are ostensibly made up a limited set of 20 standard amino acids that represent a limited range of chemical functionality compared to what can be utilized by chemists. However, nature also provides many genetically-encoded pathways to produce novel amino acids with unusual functionality that are typically not incorporated into proteins. The proposed research will utilize a pathway discovered by the Chang group that produces amino acids bearing halo, terminal alkene, and terminal alkyne functionality and will develop methods for the incorporation of these amino acids into proteins. The Chang group will further focus on a new bioinformatic strategy to identify novel pathways for biosynthesis of non-standard amino acids by using new marker enzymes known to cluster with genes involved in the biosynthesis of the amino acid itself. Further exploration of these new gene clusters can shed light on the role of non-standard amino acids in metabolism while also providing a tool to expand the diversity of proteins that can be made. This pursuit will allow graduate students and postdoctoral fellows to be broadly trained at the interface of chemistry and biology. This project will also be integrated with an outreach program to introduce K-5 students to concepts at the interface of chemistry and biology, with a focus on green chemistry.This research project focuses on the development of a bioinformatic strategy to identify novel pathways for biosynthesis of non-standard amino acids. Professor Michelle Chang and her research team will use a resistance marker to identify a wide range of gene clusters that seemingly encode the biosynthesis of new amino acids and related natural products. These gene clusters and their products will be characterized. The new knowledge will be used to expand the scope of non-standard amino acids that can be produced enzymatically. Such an approach promises to open up the possibility of introducing malleable new chemical functionalities, including alkene and alkyne groups, into targeted positions in tailored proteins. The study of the resistance enzymes will provide insight in the recognition of non-standard amino acids and will be used to design tools for residue-selective incorporation of the non-proteinogenic amino acids.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.

在化学系生命过程化学项目的支持下，加州大学伯克利分校的 Michelle Chang 教授将开发将非规范氨基酸掺入蛋白质的方法。尽管蛋白质在细胞中发挥着广泛的功能，但它们表面上是由一组有限的 20 种标准氨基酸组成，与化学家可以利用的功能相比，这些氨基酸代表了有限范围的化学功能。然而，大自然还提供了许多基因编码途径来生产具有不寻常功能的新型氨基酸，这些氨基酸通常不会掺入蛋白质中。拟议的研究将利用 Chang 小组发现的一条途径，该途径产生带有卤素、末端烯烃和末端炔烃功能的氨基酸，并将开发将这些氨基酸掺入蛋白质的方法。 Chang小组将进一步关注一种新的生物信息学策略，通过使用已知与参与氨基酸本身生物合成的基因聚集的新标记酶来确定非标准氨基酸生物合成的新途径。对这些新基因簇的进一步探索可以揭示非标准氨基酸在代谢中的作用，同时还提供扩大可制造蛋白质多样性的工具。这一追求将使研究生和博士后研究员能够在化学和生物学的交叉领域接受广泛的培训。该项目还将与一项外展计划相结合，向 K-5 学生介绍化学和生物学交叉领域的概念，重点是绿色化学。该研究项目的重点是开发生物信息学策略，以确定非标准氨基酸生物合成的新途径。米歇尔·张教授和她的研究团队将使用抗性标记来识别一系列似乎编码新氨基酸和相关天然产物生物合成的基因簇。这些基因簇及其产物将被表征。新知识将用于扩大酶法生产的非标准氨基酸的范围。 这种方法有望开辟将可延展的新化学功能（包括烯烃和炔基）引入定制蛋白质的目标位置的可能性。抗性酶的研究将为识别非标准氨基酸提供见解，并将用于设计残基选择性掺入非蛋白氨基酸的工具。该奖项反映了 NSF 的法定使命，并通过使用基金会的智力价值和更广泛的影响审查标准进行评估，被认为值得支持。