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

在化学系生命过程化学项目的支持下，哈弗福德学院的Louise Charkoudian博士和她的团队将研究如何利用细菌蛋白质组装来合成造福社会的分子。细菌已经进化出表达蛋白质组装的能力，这种组装催化制造非常复杂的分子。如果在分子水平上了解，这些蛋白质组合具有巨大的潜力，可以提供重要化学商品的可持续获取途径。这个项目将让本科生研究这些组件的进化，识别赋予这些组件功能和稳定性的特征，并参与有可能催化合成新分子的新组件的工程。这项研究将提供有关实验室从未研究过的蛋白质组装的信息。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，学习研究技能。他们还将在“共生专业发展研讨会系列”中学习如何发展专业网络以及关于不同机构和职业道路的知识。本系列课程也将向研究型大学的研究生开放。为了激发人们对科学的欣赏，促进社会各界对科学话题的讨论，查考迪安博士将开发一个名为“生物艺术贺卡”的项目，在这个项目中，学生们制作的生物艺术作品的照片会变成贺卡。第二类聚酮合成酶(PKS)是一种强大的生物催化剂，可以在反应中产生结构复杂和多样化的分子。由于缺乏获得关键的II型PKS组分的强健的异源表达系统，阻碍了体外重组II型PKS的进展。在这个项目中，生物信息学将被用来创建第二类PKS生物合成基因簇的最新目录，并根据起源的系统发育、推断的进化史和其他序列模式确定那些成熟的特征。非放线菌细菌物种将被评估为II型PKS酶的来源，这种酶可以作为稳定的、活性的蛋白质在大肠杆菌中表达。具体地说，将对酮合酶链长因子(KSCLf)、酰基载体蛋白(ACPs)和磷脂酰基转移酶(PPTase)进行表征，以确定影响其稳定性和功能的特征。将探索第二类PKS ACP与不同PKS之间的PPTase的兼容性。该奖项反映了NSF的法定使命，并已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 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.