Synthetic Biology Approaches to Selective Amino Acid Modification and Incorporation

在化学学部生命过程化学项目的支持下，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整合到蛋白质中的方法。尽管蛋白质在细胞中发挥着广泛的功能，但表面上看，它们是由一组有限的20种标准氨基酸组成的，与化学家可以利用的氨基酸相比，它们代表的化学功能范围有限。然而，大自然也提供了许多遗传编码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纳入蛋白质中。拟议的研究将利用Chang小组发现的产生具有halo，末端烯烃和末端炔功能的氨基酸的途径，并将开发将这些氨基酸结合到蛋白质中的方法。Chang团队将进一步关注一种新的生物信息学策略，通过使用已知的与氨基酸自身生物合成相关基因聚类的新标记酶，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，同时也为扩大可制造蛋白质的多样性提供了一种工具。这种追求将使研究生和博士后在化学和生物学的界面上得到广泛的训练。该项目还将与一项外展计划相结合，向K-5年级的学生介绍化学和生物学的概念，重点是绿色化学。本研究项目的重点是发展一种生物信息学策略，以确定非标准氨基酸的生物合成新途径。张雪儿教授和她的研究小组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的生物合成的基因簇。这些基因簇及其产物将被表征。这一新知识将用于扩大可酶法产生的非标准氨基酸的范围。这种方法有望在定制蛋白质的目标位置引入可延展的新化学功能，包括烯烃和炔基。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氨基酸残基选择性结合的工具。该奖项反映了美国国家科学基金会的法定使命，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.