CAREER: Tackling the Solvent-Stabilizer Co-contamination by Propanotrophic Bacteria with Catalytically Versatile Di-iron Monooxygenases

成千上万的含水层被氯代脂肪烃（CAH）溶剂和稳定剂（如1,4-二氧六环）的混合物污染。迄今为止，还没有一种单一的补救策略能够同时处理这两种类型的污染物。为了解决这个问题，将开发一种创新的生物处理方法，利用在丙烷上生长的细菌，这种细菌能够同时降解CAHs和1,4-二恶烷。这种方法的发展需要分离出能够在各种野外条件下进行有效治疗的各种细菌菌株。这种新颖的修复策略在精确定位特定污染地点方面具有潜在的变革性。这种方法的另一个好处是，与其他可用的补救策略相比，它对环境的破坏较小。通过实验室实验和与利益相关者的实践培训，研究者将培养下一代环境修复专业人员。招收来自不同背景的学生将扩大这些领域的参与。生物修复通常是首选的地下水污染处理方法，因为它使用自然方法以相对较低的成本降解污染物。然而，这种方法的成功高度依赖于特定的物理化学特征和给定地点的本地微生物群，导致特定地点的解决方案适用性狭窄。为了提高生物修复的有效性，PI将建立一个具有强大降解能力和环境适应性的propanotrophic分离株和联合体的集合。该策略依靠传统培养方法和最先进的生物技术（如宏基因组学和单细胞基因组分析）的协同作用，在分子、细胞和微生物群落水平上推进对彗星代谢生物降解的理解。本研究的重点是丙烷诱导的可溶性二铁单加氧酶（SDIMOs）作为多功能生物催化剂，具有独特的降解多种氯化溶剂和1,4-二氧六环的能力。将鉴定新的SDIMOs，并在异源表达克隆和敲除突变体中表征其催化能力和动力学。这些见解将用于完善SDIMOs的分类，以便设计和解释组学和其他分子生物学工具，以利用从不同污染栖息地获得的proprotrophic consortium之间的代谢相互作用。最后，将采用实验规模的微观分析来评估和比较传统和基于诊断的生物修复策略的可行性和可靠性，以推进其原位应用。这项研究将积极与核心教育目标相结合，以培养下一代修复专业人员，以解决历史上受污染的纽约-新泽西-康涅狄格三州地区日益增长的需求。利用该地区规模庞大的修复行业，PI将吸引各级学生参加各种校内和校外活动，例如与环境专家的客座讲座，与该领域校友的专业研讨会，大学赞助的研究项目，以及与该地区私人和非营利修复组织的实习和交流机会。这些经历将为学生提供在该领域进行应用研究和培训的关键途径，并培养长期的联系，这是职业成功的关键催化剂。此外，PI还将吸引两年制大学和地方高中中代表性不足的群体中的积极学生参加科学研讨会、实验室参观和兼职研究机会。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。

Thousands of aquifers are contaminated with mixtures of chlorinated aliphatic hydrocarbon (CAH) solvents and stabilizers like 1,4-dioxane. To date, there is no single remediation strategy capable of treating both types of pollutants simultaneously. To tackle the problem, an innovative biological treatment will be developed that employs bacteria growing on propane that are capable of simultaneously degrading both CAHs and 1,4-dioxane. Development of this approach requires the isolation of a diverse collection of bacterial strains capable of effective treatment under various field conditions. This novel remediation strategy is potentially transformative in its precision targeting for specific contaminated sites. An added benefit of this approach results from it being less disruptive to the environment compared to other available remedial strategies. Through laboratory experimentation and hands-on training with stakeholders, the investigator will develop the next generation of environmental remediation professionals. Recruitment of students from diverse backgrounds will broaden participation in these fields.Bioremediation is frequently a preferred groundwater contamination treatment method because it uses a natural approach to degrade pollutants at relatively low cost. However, the success of this approach is highly dependent on the specific physiochemical characteristics and the indigenous microbiota of a given site, resulting in site-specific solutions with narrow applicability. To increase the effectiveness of bioremediation, the PI will establish a collection of propanotrophic isolates and consortia exhibiting robust degradation capabilities and environmental adaptabilities. This strategy relies on the synergy of conventional cultivation approaches and state-of-the-art biotechnologies such as metagenomics and single-cell genomic analysis to advance understanding of cometabolic biodegradation at the molecular, cellular, and microbial community levels. This research focuses on propane-inducible soluble di-iron monooxygenases (SDIMOs) as versatile biocatalysts, uniquely capable of degrading a variety of chlorinated solvents and 1,4-dioxane. Novel SDIMOs will be identified and their catalytic capacities and kinetics will be characterized in heterologous expression clones and knockout mutants. These insights will be used to refine the categories of SDIMOs to enable the design and interpretation of omics and other molecular biological tools to exploit the metabolic interactions among propanotrophic consortia obtained from different polluted habitats. Finally, bench-scale microcosm assays will be used to evaluate and compare the feasibility and reliability of conventional and diagnosis-based bioremediation strategies to advance their in situ applications. This research will be actively integrated with a core educational goal to cultivate the next generation of remediation professionals needed to address growing demand in the historically polluted New York-New Jersey-Connecticut tristate area. Taking advantage of the sizeable remediation industry in this area, the PI will engage students of all levels with a diverse range of on- and off-campus activities, such as guest lectures with environmental experts, professional workshops with alumni in the field, and University-sponsored research projects, as well as internships and networking opportunities with private and non-profit remediation organizations in the region. These experiences will give students critical access to applied research and training in the field, and foster long-lasting connections that are key catalysts for career success. Further, the PI will attract motivated students from underrepresented groups at two-year colleges and local high schools to take part in scientific seminars, laboratory tours, and part-time research opportunities.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.