Collaborative Research: Droplet-based selection to improve aflatoxin detoxification

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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.