Collaborative Research: Droplet-based selection to improve aflatoxin detoxification

真菌是可以在谷物和水果上自然生长的微生物。当真菌生长时，有些会产生毒素，当食用时会损害我们的神经系统，肝脏和尿道。因此，真菌污染对农业造成了相当大的经济负担，因为没有成本效益高的方法来去除它们产生的毒素。一些被称为“解毒剂”的细菌已经被鉴定出可以降解这些毒素。然而，天然存在的解毒剂不足以有效地解决这个问题。这项研究的目的是了解解毒剂如何降解真菌毒素，最终目的是保护食物供应。这一目标将使用高度新颖的单液滴分选平台来实现。将从原始菌株中突变出除臭剂，并测试其降解液滴中真菌毒素的能力。通过监测和分类大量的液滴，解毒剂变体将被迅速识别出效果最好。将对这些菌株进行研究，以了解毒素降解过程。这项研究的成功完成将导致解决真菌毒素治疗的新策略。它还将导致开发一种新的系统方法，用于丰富细菌解毒剂的一系列应用。该项目将通过培训未来的环境工程研究人员来提高国家的STEM劳动力，并通过公众教育和食品安全和污染的宣传来提高科学素养。该项目的目标是了解细菌用于解毒污染我们食物的有害真菌毒素的机制。一个高通量的基于液滴的平台将被用来表征和分类携带解毒酶的细菌细胞，这些解毒酶集中在黄曲霉毒素上，黄曲霉毒素是一种无处不在的有害毒素，污染了许多食品和饲料资源。该研究设计利用黄曲霉毒素的天然荧光进行高通量传感，以创建解毒剂突变体库。这些突变体将在随后的基因组和转录组学研究中进行筛选，以确定参与解毒过程的基因。这个合作项目结合了定量生物学、基于液滴的分选技术以及基因组学和转录组学工具，为揭示黄曲霉毒素降解机制提供了一种简化的策略。这些信息对于开发保护食品供应免受这种有毒化合物影响的解决方案至关重要。从这项研究中获得的知识有望为改进解毒菌株的系统工程开辟新的途径。该项目将有助于在科学和工程的接口跨学科专家的培训和教育，从而提高国家的STEM劳动力。研究结果、方法和工具将通过对食品污染的公共健康和经济影响的宣传传播给科学界和公众，从而提高科学素养。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。

Fungi are microorganisms that can grow naturally on cereals and fruits. When fungi grow, some can produce toxins that can damage our nervous system, liver, and urinary tract when consumed. Fungal contamination thus poses considerable economic burdens on agriculture, as there are no cost-effective approaches to removing the toxins they produce. Some species of bacteria called ‘detoxifiers’ have been identified that can degrade these toxins. However, naturally occurring detoxifiers are not efficient enough to address this problem. The aim of this research is to understand how detoxifiers degrade fungal toxins with the ultimate goal of protecting the food supply. This goal will be achieved using a highly novel single droplet sorting platform. Detoxifiers will be mutated from the original strains and tested for the ability to degrade fungal toxins in the droplet. By monitoring and sorting large numbers of droplets, detoxifier variants will be rapidly identified that work best. These strains will be investigated to understand the toxin degradation process. Successful completion of this research will lead to new strategies for addressing fungal toxin treatment. It will also lead to the development of a novel systematic method for enriching bacterial detoxifiers for a range of applications. This project will improve the Nation’s STEM workforce through the training of future researchers in environmental engineering and will also increase scientific literacy through public education and outreach on food safety and contamination.The goal of this project is to understand the mechanisms bacteria use to detoxify harmful fungal toxins that contaminate our food. A high-throughput droplet-based platform will be used to characterize and sort bacterial cells that carry detoxification enzymes focusing on aflatoxin—a ubiquitous, harmful toxin that contaminates many food and feed resources. The research design exploits the natural fluorescence of aflatoxin for high throughput sensing to create a library of detoxifier mutants. These mutants will be screened in subsequent genomic and transcriptomic studies to identify genes involved in the detoxification process. This collaborative project combines quantitative biology, droplet-based sorting technology, and genomic and transcriptomic tools to offer a streamlined strategy for uncovering the mechanisms of aflatoxin degradation. This information is critical to develop solutions for protecting the food supply from this toxic compound. Knowledge gained from this research holds promise to open up new avenues for systematic engineering of improved detoxifying strains. This project will contribute to the training and education of interdisciplinary experts at the interface of science and engineering, thus improving the Nation’s STEM workforce. The findings, methods, and tools will be disseminated to the scientific community, as well as the general public through outreach on public health and economic impact of food contamination, thus improving scientific literacy.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.