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

职业：N-羟化酶生物催化剂的表征和进化：催化和修复金属污染的解决方案

• 批准号: 2236643
• 负责人: Lesley-Ann Giddings
• 金额: $ 61.54万
• 依托单位: Smith College
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-03-01 至 2028-02-29
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2236643&HistoricalAwards=false
• 关键词: CAREER; Characterization; Evolution; hydroxylase; Biocatalysts

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.

大多数细菌会产生与铁原子结合的小分子。这些分子称为铁载体。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污染的铜矿场中搜索具有新型重金属结合活性的铁载体。一旦发现，将确定并设计了负责产生这些金属结合分子的酶以增强功能。该项目将吸引包括有色女性在内的本科妇女研究。在马萨诸塞州斯普林菲尔德的斯普林菲尔德的城市青年STEM富集计划的年度研讨会的形式还将举行社区宣传。由于烷基二氨酸N-羟化酶单加氧酶（NMOS）参与了铁载体生物合成，因此新型的铁载体可能是独特的NMO的产物。可以预期，参与新型铁载体生物合成的NMO具有更广泛的底物特异性，使其成为识别和开发新的工业生物催化剂的目标。例如，羟胺，化学和制药行业中有价值的构建块是从羟氨酸型铁载体生物合成中的单一步骤中酶做出的。该项目将表征新识别的NMO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的法定任务，并通过使用基金会的智力优点和更广泛的影响来审查标准，被认为通过评估来获得珍贵的支持。