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

这一行动为NSF 2019财年生物学博士后研究奖学金-使用生物收藏进行研究-提供资金。该奖学金支持将以创新方式利用生物收藏的研究员的研究和培训。被称为蓝藻的光合作用细菌产生各种具有不同寻常的化学结构的有机分子，包括含有氯和溴的卤化分子，这些分子是使用称为卤化酶的酶来产生的。这种酶是不寻常的，因为它们可以在碳原子上形成其他酶无法使用的化学键。对卤素酶的更好理解将导致对化学的新见解。为了实现这一目标，这位研究员将研究一种新发现的卤素酶，这种酶能够将卤素原子连接到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，并将其应用于开发新的化学合成方法，以生产具有工业重要性的分子。这位研究员将参与公共宣传活动，包括互动的博物馆展品，以强调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序列数据从据称的蓝藻标本中识别新的卤代酶。使用基于聚合酶链式反应的方法，这位研究员将识别含有脂肪酸卤化酶同源物的菌株，并通过全基因组测序进行后续验证。这位研究员将克隆、异源表达、纯化和功能鉴定来自ALICE收藏的新的同源物，以及来自其他培养收藏的同源物，以确定酶选择性的遗传决定因素，并探索进化和酶活性之间的关系。这位研究员还将使用液相色谱和质谱学来鉴定大学收藏中的物种产生的新的卤代化合物。这位研究员将分离和表征这些化合物，目标是将新的含有卤代酶的生物合成基因簇与它们产生的分子联系起来。通过对蓝藻中脂肪酸卤化酶的功能和遗传多样性的表征，这项工作将证明新的卤化酶在化学合成中用于碳活化的用途。在进行这项研究时，该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，该研究员将对他们进行培训和指导，并将在化学和生物领域的跨学科数据集分析方面建立新的合作。该奖项反映了NSF的法定使命，并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。

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.