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

本行动资助NSF 2019年度生物学博士后研究奖学金，利用生物馆藏进行研究。该奖学金支持研究和培训将以创新的方式利用生物收集的研究员。被称为蓝细菌的光合细菌产生各种具有不同化学结构的有机分子，包括含氯和溴的卤化分子，这些分子是通过称为卤化酶的酶产生的。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利用的化学键。对卤化酶的更好理解将导致化学上的新见解。为了实现这一目标，他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脂肪酸分子上。该项目将为理解一种新型碳活化酶奠定基础，并应用于开发新的化学合成方法来生产工业上重要的分子。该研究员将参与公众宣传，包括互动博物馆展览，以突出微生物学在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序列数据从蓝藻的LEGE收集中识别新的卤化酶。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LEGE菌株，并通过全基因组测序进行后续验证。该研究员将克隆、异种表达、纯化和功能表征来自LEGE收集的新同源物，以及来自其他培养收集的同源物，以确定酶选择性的遗传决定因素，并探索进化与酶活性之间的关系。该研究员还将使用液相色谱法和质谱法来鉴定LEGE收集的物种产生的新卤化化合物。他将对这些化合物进行分离和结构表征，目标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，本工作将记录新的卤化酶在化学合成中碳活化的效用。在进行这项研究时，研究员将接受生物信息学、化学合成、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，该研究员将培训和指导本科生，并将为化学和生物学跨学科数据集的分析建立新的合作关系。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。

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.