CAREER: Thermal proteome profiling applications for in-situ bioreporting of contaminant degradation in soils

新出现的污染物，通常被称为新出现的关注污染物(CECs)，是越来越多地在土壤、地表水系统和地下水含水层中检测到的人为化学污染物。在美国，CEC对国家的饮用水资源构成了重大威胁。土壤中的微生物通过将人为化学污染物分解成无害和无毒的副产物，在饮用水水源保护中发挥着关键作用。然而，我们培养土壤微生物和表征/利用负责降解CECs的酶的能力仍然难以捉摸。这个职业项目的总体目标是促进对微生物酶-土壤相互作用的基本理解，目标是开发和验证基于蛋白质组的分析和技术，以探索土壤中CECs的微生物降解。为了推进这一目标，首席研究员建议将土壤特性、土壤活性酶提取、预测建模和热蛋白质组图谱(TPP)相结合，以1)探测和揭示土壤中CECs的微生物衰减，2)检测和发现能够降解土壤中CECs的新型酶。该项目的成功完成将使社会受益，因为它将产生关于CEC微生物降解途径的新的基本知识，以支持开发和部署保护饮用水水源的天然和工程解决方案。还将通过学生教育和培训，包括指导堪萨斯大学的一名研究生和四名本科生，为社会带来更多好处。新出现的问题(CEC)的污染物威胁到该国的饮用水来源和获得清洁淡水的途径。了解和利用土壤、地表水系统和地下含水层中微生物群落对CECs的降解可能会带来更大的水源保护。这个职业项目的目标是利用先进的蛋白质组学技术，包括热蛋白质组图谱(TPP)，调查和揭示土壤中微生物群落对CECs的降解途径。这种创新的方法利用在药物发现中广泛开发和使用的方案和分析来识别药物的蛋白质靶标。然而，将TPP应用于环境修复问题的一个障碍是缺乏关于活性酶如何与复杂的土壤基质相互作用的知识，包括评估和量化从土壤中提取活性污染物降解酶的可靠框架。这项研究的具体目标是：1)探索和揭示污染物降解酶与复杂土壤基质的相互作用，2)基于吸附等温线、蛋白水解势和机器学习方法开发土壤酶提取效率的预测模型，以及3)整合模型预测和先进的“基因组学”技术(例如，元基因组学和元转录组学)，以验证TPP作为(I)识别参与污染物降解的酶/蛋白质和(Ii)土壤中污染物降解的原位生物报告的平台的使用。该项目的成功完成有可能产生变革性的影响，通过产生关于酶-土壤相互作用的新的基础知识，以支持开发和部署天然和工程解决方案，以保护饮用水水源免受CEC的影响。为了落实这一职业项目的教育和培训目标，首席调查员(PI)建议开发课堂和基于研究的教育模块，以增加大专转学学生的动力和留住他们。本课程将与堪萨斯大学新兴学者项目提供的研究机会相结合。大专转校生将通过一个由八所堪萨斯州社区学院组成的网络进行识别。此外，PI计划将教育模块分发给罗斯-胡尔曼理工学院，这是一所主要是本科生的机构，以招收和吸引堪萨斯大学环境工程研究的本科生。这一奖项反映了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。

Emerging contaminants, commonly referred to as contaminants of emerging concern (CECs), are anthropogenic chemical pollutants that are increasingly being detected in soils, surface water systems, and groundwater aquifers. In the United States, CECs pose a significant threat to the Nation’s drinking water resources. Microorganisms in soils play a critical role in the protection of drinking water sources by degrading anthropogenic chemical pollutants into harmless and nontoxic byproducts. However, our ability to grow soil microorganisms and characterize/harness the enzymes responsible for the degradation of CECs has remained elusive. The overarching goal of this CAREER project is to advance the fundamental understanding of microbial enzyme-soil interactions with the goal of developing and validating proteomic-based assays and techniques to probe the microbial degradation of CECs in soils. To advance this goal, the Principal Investigator proposes to combine soil characterization, active enzyme extraction from soils, predictive modeling, and thermal proteome profiling (TPP) to 1) probe and unravel the microbiological attenuation of CECs in soils and 2) detect and discover novel enzymes that can degrade CECs in soils. The successful completion of this project will benefit society through the generation of new fundamental knowledge on CEC microbial degradation pathways to support the development and deployment of natural and engineered solutions to protect drinking water sources. Additional benefits to society will be achieved through student education and training including the mentoring of a graduate student and four undergraduate students at the University of Kansas. Contaminants of emerging concerns (CECs) threaten the Nation’s drinking water sources and access to clean and fresh water. Understanding and harnessing the degradation of CECs by microbial communities in soils, surface water systems and groundwater aquifers could lead to greater source water protection. The goal of this CAREER project is to investigate and unravel the degradation pathways of CECs by microbial communities in soils using advanced proteomic techniques including thermal proteome profiling (TPP). This innovative approach leverages protocols and assays developed and used extensively in drug discovery to identify a pharmaceutical's protein target. However, a barrier to applying TPP to environmental remediation problems is a lack of knowledge of how active enzymes interact with complex soil matrices, including a robust framework to assess and quantify the extraction of active contaminant-degrading enzymes from soils. The specific objectives of this research are to: 1) probe and unravel the interactions of contaminant-degrading enzymes with complex soil matrices, 2) develop predictive models of soil-enzyme extraction efficiency based on sorption isotherms, proteolytic potentials, and machine-learning approaches, and 3) integrate model predictions and advanced “omic” techniques (e.g., metagenomics and meta transcriptomics) to validate the use of TPP as a platform for (i) identifying enzymes/proteins involved in contaminant degradation and (ii) in-situ bioreporting of contaminant degradation in soils. The successful completion of this project has the potential for transformative impact through the generation of new fundamental knowledge on enzyme-soil interactions to support the development and deployment of natural and engineered solutions to protect drinking water sources from CECs. To implement the educational and training goals of this CAREER project, the Principal Investigator (PI) proposes to develop classroom and research-based education modules to increase the motivation and retention of junior college transfer students. This coursework will be paired with research opportunities provided by the University of Kansas Emerging Scholars Program. Junior college transfer students will be identified through a network of eight Kansas community colleges. In addition, the PI plans to disseminate the education modules to Rose-Hulman Institute of Technology, a primarily undergraduate institution, to recruit and engage undergraduate students in environmental engineering research at the University of Kansas.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.