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，加州大学伯克利分校的Michelle Chang教授将开发将非规范氨基酸整合到蛋白质中的方法。虽然蛋白质在细胞中执行广泛的功能，但它们表面上是由20个标准氨基酸组成的有限集合，与化学家可以利用的相比，这些氨基酸代表了有限的化学功能范围。然而，大自然也提供了许多遗传编码的途径来产生具有不寻常功能的新氨基酸，这些氨基酸通常不会被整合到蛋白质中。这项拟议的研究将利用Chang小组发现的一条途径，产生具有卤代、末端烯烃和末端炔官能团的氨基酸，并将开发将这些氨基酸结合到蛋白质中的方法。Chang团队将进一步专注于一种新的生物信息学策略，通过使用新的标记酶来确定非标准氨基酸生物合成的新途径，新的标记酶与参与氨基酸生物合成的基因聚集在一起。对这些新基因簇的进一步探索可以阐明非标准氨基酸在新陈代谢中的作用，同时也提供了一种工具来扩大可以制造的蛋白质的多样性。这一追求将使研究生和博士后研究员在化学和生物方面得到广泛的培训。这个项目还将与一个推广计划相结合，向K-5学生介绍化学和生物界面的概念，重点是绿色化学。本研究项目重点是开发一种生物信息学策略，以确定非标准氨基酸的生物合成的新途径。米歇尔·张教授和她的研究团队将使用一种抗性标记来识别一系列似乎编码新氨基酸和相关天然产品生物合成的基因簇。这些基因簇及其产物将被表征。这项新知识将用于扩大酶法生产的非标准氨基酸的范围。这种方法有望为将可延展的新化学官能团，包括烯烃和炔基，引入到定制蛋白质的目标位置提供可能性。对抗性酶的研究将为识别非标准氨基酸提供洞察力，并将用于设计工具，选择性地纳入非蛋白生成氨基酸。这一奖项反映了NSF的法定使命，并通过使用基金会的智力优势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.