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，来自哈弗福德学院的Louise Charkoudian博士和她的团队将研究如何利用细菌蛋白质的组装来合成有益于社会的分子。细菌已经进化出了表达蛋白质组合的能力，这种能力催化了非常复杂分子的制造。如果在分子水平上理解，这些蛋白质组合具有巨大的潜力，可以为重要的化学商品提供可持续的途径。该项目将吸引本科生研究这些组合的进化，识别赋予这些组合功能和稳定性的特征，以及具有催化新分子合成潜力的新组合的工程设计。这项研究将提供从未在实验室中研究过的蛋白质组合的信息。还将开发促进未来发现和工程工作的工具。从事本研究的本科生将获得科学知识和研究技能。他们还将在“共生专业发展系列研讨会”中学习如何发展专业网络，了解不同的机构和职业道路。这个系列也将开放给研究型大学的研究生。为了激发科学鉴赏和促进社区参与科学话题的讨论，Charkoudian博士将开展一个“生物艺术贺卡”项目，将学生制作的生物艺术作品的照片变成贺卡。II型聚酮合成酶（pks）是一种强大的生物催化剂，可以产生结构复杂和多样的分子。由于缺乏获得关键II型PKS成分的强大的异源表达系统，在体外重建II型PKS的进展受到阻碍。在这个项目中，生物信息学将用于创建II型PKS生物合成基因簇的更新目录，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。非放线菌细菌种类将被评估为II型PKS酶的来源，这种酶可以在大肠杆菌中作为稳定的活性蛋白表达。具体来说，酮合酶链长因子（KSCLFs）、酰基载体蛋白（ACPs）和磷酸antetheinyl转移酶（PPTases）将被表征，以确定影响其稳定性和功能的特征。II型PKS ACPs与跨不同PKS的pptase的兼容性将被探讨。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 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.