Postdoctoral Fellowship in Biology FY 2019: The Diversity of a New Carbon-Activating Halogenase from Cyanobacteria

这项行动资助了2019财年NSF生物学博士后研究奖学金，使用生物收集的研究。该研究金支持研究员的研究和培训，以创新的方式利用生物收藏。被称为蓝细菌的光合细菌产生各种具有不寻常化学结构的有机分子，包括卤化分子，其含有氯和溴，并使用称为卤化酶的酶产生。这种酶很不寻常，因为它们可以在碳原子上形成其他酶无法使用的化学键。更好地理解卤化酶将导致化学的新见解。为了实现这一目标，这位研究员将研究一种新发现的卤化酶，这种酶能够将卤素原子附着在称为脂肪酸的分子上。该项目将为理解一种新型的碳活化酶奠定基础，并应用于化学中新的合成方法的开发，以生产工业上重要的分子。该研究员将参与公共宣传活动，包括互动式博物馆展览，以突出微生物学在人类日常生活中的重要性。在与葡萄牙波尔图大学的国际合作中，这项研究将使用DNA序列数据从乐歌收集的蓝藻中鉴定新的卤化酶。使用基于PCR的方法，该研究员将鉴定含有脂肪酸卤化酶同源物的乐歌菌株，并通过全基因组测序进行后续验证。该研究员将克隆，异源表达，纯化和功能特性的新同系物从乐歌收集，除了同系物从其他文化收藏，以确定酶的选择性的遗传决定因素，并探讨进化和酶活性之间的关系。该研究员还将使用液相色谱法和质谱法来鉴定乐歌收集的物种产生的新的卤代化合物。该研究员将分离和结构表征这些化合物，目的是将新的含卤素酶的生物合成基因簇与它们产生的分子连接起来。通过表征蓝藻中脂肪酸卤化酶的功能和遗传多样性，这项工作将记录新的卤化酶在化学合成中用于碳活化的效用。在进行这项研究时，研究员将接受生物信息学、化学合成和天然产物分离和表征方面的培训。这项研究将在本科生的帮助下进行，研究员将对本科生进行培训和指导，并将为化学和生物学的跨学科数据集分析建立新的合作关系。该奖项反映了NSF的法定使命，并被认为值得通过使用基金会的智力价值和更广泛的影响审查标准进行评估来支持。

This action funds an NSF Postdoctoral Research Fellowship in Biology for FY 2019, Research Using Biological Collections. The fellowship supports research and training of the fellow that will utilize biological collections in innovative ways. Photosynthetic bacteria known as cyanobacteria produce a variety of organic molecules with unusual chemical structures, including halogenated molecules, which contain chlorine and bromine and are produced using enzymes called halogenases. The enzymes are unusual because they can form chemical bonds at carbon atoms that other enzymes cannot use. A better understanding of halogenases will lead to new insights in chemistry. Towards that goal, the fellow will study a newly discovered type of halogenase that is able to attach halogen atoms to molecules called fatty acids. This project will lay the groundwork for understanding a new type of carbon-activating enzyme, with applications for the development of new synthetic methods in chemistry to produce industrially important molecules. The fellow will engage in public outreach, including interactive museum exhibits, to highlight the importance of microbiology in every-day human life.In an international collaboration with the University of Porto, Portugal, this research will use DNA sequence data to identify new halogenases from the LEGE collection of cyanobacteria. Using a PCR-based approach, the fellow will identify LEGE strains that contain homologs of a fatty acid halogenase, with follow-up validation by whole genome sequencing. The fellow will clone, heterologously express, purify, and functionally characterize the new homologs from the LEGE collection, in addition to homologs from other culture collections, to identify genetic determinants of enzyme selectivity and explore the relationships between evolution and enzyme activity. The fellow will also use liquid chromatography and mass spectrometry to identify new halogenated compounds produced by species in the LEGE collection. The fellow will isolate and structurally characterize these compounds with the goal of connecting new halogenase-containing biosynthetic gene clusters to the molecules they produce. By characterizing the functional and genetic diversity of fatty acid halogenases in cyanobacteria, this work will document the utility of new halogenases for carbon activation in chemical synthesis. In performing this research, the fellow will receive training in bioinformatics, chemical synthesis, and natural products isolation and characterization. This research will be performed with assistance from undergraduates, whom the fellow will train and mentor, and it will build new collaborations for the analysis of interdisciplinary datasets in chemistry and biology.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.