RUI: Biochemical Comparison of Type II Polyketide Biosynthetic Enzymes Across Phyla for Expanded Access to Chemical Diversity

RUI：跨门的 II 型聚酮化合物生物合成酶的生化比较，以扩大化学多样性的获取

• 批准号: 2201984
• 负责人: Louise Charkoudian
• 金额: $ 44.7万
• 依托单位: Haverford College
• 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: 2022-12-15 至 2026-02-28
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2201984&HistoricalAwards=false
• 关键词: RUI; Biochemical; Comparison; Type; II

With the support of the Chemistry of Life Processes Program in the Division of Chemistry, Dr. Louise Charkoudian and her team from Haverford College will investigate how assemblies of bacterial proteins can be leveraged to synthesize molecules that benefit society. Bacteria have evolved the ability to express protein assemblies that catalyze the manufacture of very complicated molecules. If understood at the molecular level, these protein assemblies hold tremendous potential to provide sustainable access to important chemical commodities. This project will engage undergraduate students in the study of the evolution of these assemblies, in identifying the features that confer function and stability upon these assemblies, and in the engineering of new assemblies that have the potential to catalyze the synthesis of novel molecules. The research will provide information about protein assemblies that have never been studied in the laboratory. Tools to facilitate future discovery and engineering efforts will also be developed. The undergraduate students engaged in this research will gain scientific knowledge and learn research skills. They will also learn how to develop professional networks and about different institutions and career paths in a “Symbiotic Professional Development Seminar Series”. This series will also be open to graduate students from research universities. To spark scientific appreciation and foster engagement of the community in discussions of scientific topics, Dr. Charkoudian will develop a “BioArt Greeting Cards” project in which photographs of student-made pieces of biological art are turned into greeting cards.Type II polyketide synthases (PKSs) are powerful biocatalysts in reactions that produce structurally complex and diverse molecules. Progress towards reconstituting type II PKSs in vitro has been stymied by the lack of a robust heterologous expression system for obtaining key type II PKS components. In this project, bioinformatics will be used to create an updated catalogue of type II PKS biosynthetic gene clusters and to identify those ripe for characterization based on phylogeny of origin, inferred evolutionary history, and other sequence patterns. Non-actinomycete bacterial species will be evaluated as a source of type II PKS enzymes that can be expressed in E. coli as stable, active proteins. Specifically, ketosynthase chain length factors (KSCLFs), acyl carrier proteins (ACPs) and phosphopantetheinyl transferases (PPTases) will be characterized to determine the features that influence their stability and function. The compatibility of type II PKS ACPs with PPTases across different PKSs will be explored.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.

在化学过程中的化学过程中的支持下，路易丝·查库尼亚（Louise Charkoudian）博士及其来自哈弗福德学院（Haverford College）的团队将调查如何利用细菌蛋白的组装来综合使社会受益的分子。细菌已经发展出表达蛋白质组件的能力，从而催化非常复杂的分子的生产。如果在分子水平上理解，这些蛋白质组件具有巨大的潜力，可以为重要的化学商品提供可持续的机会。该项目将吸引本科生研究这些组装的演变，以确定这些组装上的会议功能和稳定性的特征，以及在具有催化新分子综合的新组装工程中的工程。该研究将提供有关从未在实验室研究的蛋白质组件的信息。还将开发促进未来发现和工程工作的工具。从事这项研究的本科生将获得科学知识并学习研究技能。他们还将学习如何在“共生专业发展研讨会系列”中开发专业网络以及有关不同机构和职业道路。该系列还将向研究大学的研究生开放。为了激发社区的科学欣赏和促进科学主题的讨论，Charkoudian博士将开发一个“生物Art贺卡”项目，在该项目中，学生制作的生物学艺术品的照片变成了贺卡。型型Polyketide Synthases（PKSS）是有力的生物催化剂，这些反应是产生结构上复杂和多种分类分类的反应。由于缺乏可获得关键的II型PKS成分的强大异源表达系统，因此在体外重新建立II型PKS的进展受到阻碍。在该项目中，生物信息学将用于创建II型PKS Biosynthetic Gene基因簇的更新目录，并根据原产系统，推断的进化历史和其他序列模式来识别那些成熟的表征。非肌动菌细菌将被评估为II型PKS酶的来源，可以在大肠杆菌中以稳定的活性蛋白质表示。具体而言，将对酮合酶链长度因子（KSCLF），酰基载体蛋白（ACP）和磷酸乙烯基转夹（PPTase）进行表征，以确定影响其稳定性和功能的特征。将探讨II型PKS ACP与PPTase跨不同PKS的兼容性。该奖项反映了NSF的法定任务，并使用基金会的知识分子优点和更广泛的影响审查标准，被认为是通过评估来获得的支持。