CAREER: Characterization and Evolution of N-hydroxylase Biocatalysts: Solutions to Catalysis and Remediating Metal Pollution

大多数细菌产生与铁原子结合的小分子。这些分子被称为铁载体。许多铁载体也可以结合有毒的重金属。该项目将在佛蒙特州一个受污染的铜矿中寻找具有新的重金属结合活性的铁载体。一旦发现，负责产生这些金属结合分子的酶将被识别和工程化以增强能力。 该项目将吸引包括有色人种在内的女大学生参与研究。也将有社区外展的气候正义的年度研讨会的形式，涉及斯普林菲尔德的青年STEM富集计划在斯普林菲尔德，MA城市联盟。保守的序列基序中确定的新铁载体的基因分离的金属污染的环境将被确定和表征。由于烷基二胺N-羟化酶单加氧酶（NMOs）参与铁载体的生物合成，新的铁载体可能是独特的NMOs的产物。预计参与新型铁载体生物合成的NMOs具有更广泛的底物特异性，使其成为鉴定和开发新的工业生物催化剂的靶标。例如，羟胺，化学和制药工业中有价值的结构单元，在异羟肟酸盐铁载体的生物合成中由伯胺在单一步骤中酶促制备。 该项目将表征新发现的NMOs的选择性，并进化这些酶，提高其产量和产品多样性。该奖项反映了NSF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。

Most bacteria create small molecules that bind to iron atoms. These molecules are called siderophores. Many siderophores can also bind toxic heavy metals. This project will search for siderophores with novel heavy metal binding activities in a polluted copper mine site in Vermont. Once found, the enzymes responsible for producing these metal-binding molecules will be identified and engineered for enhanced capabilities. The project will engage undergraduate women, including women of color, in research. There will also be community outreach in the form of annual workshops on climate justice involving the Urban League of Springfield’s Youth STEM enrichment program in Springfield, MA.Conserved sequence motifs identified in genes for novel siderophores isolated from a metal-polluted environment will be identified and characterized. Since alkyl diamine N-hydroxylase monooxygenases (NMOs) are involved in siderophore biosynthesis, the novel siderophores will likely be the product of unique NMOs. It is expected that NMOs involved in novel siderophore biosynthesis have broader substrate specificity, making them a target for identifying and developing new industrial biocatalysts. For example, hydroxylamines, valuable building blocks in the chemical and pharmaceutical industries, are enzymatically made in a single step from primary amines in the biosynthesis of hydroxamate siderophores. This project will characterize the selectivity of newly-identified NMOs and evolve these enzymes, increasing their yield and product diversity.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.