Synthetic Biology Approaches to Selective Amino Acid Modification and Incorporation

在化学系生命过程化学计划的支持下，来自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。虽然蛋白质在细胞中执行广泛的功能，但它们表面上由20种标准氨基酸组成，与化学家可以利用的化学功能相比，这些氨基酸代表了有限的化学功能。然而，自然界也提供了许多遗传编码的途径，以产生具有不寻常功能的新型氨基酸，这些氨基酸通常不会被掺入蛋白质中。拟议的研究将利用Chang小组发现的一种途径，该途径产生带有卤素，末端烯烃和末端炔官能的氨基酸，并将开发将这些氨基酸掺入蛋白质的方法。Chang小组将进一步专注于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簇合的新标记酶来识别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，同时也提供了一种扩大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在化学和生物学的接口得到广泛的培训。本项目还将与一个外展计划相结合，向K-5学生介绍化学和生物学界面的概念，重点是绿色化学。本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。Michelle Chang教授和她的研究团队将使用一种抗性标记来鉴定一系列基因簇，这些基因簇似乎编码新氨基酸和相关天然产物的生物合成。这些基因簇及其产物将被表征。新的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官能团（包括烯基和炔基）引入定制蛋白质的目标位置的可能性。对抗性酶的研究将为非标准氨基酸的识别提供见解，并将用于设计非蛋白质氨基酸残基选择性掺入的工具。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。

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.