Many natural microorganisms, such as bacteria and fungi, can be used to degrade toxic pollutants and remediate contaminated sites. These microorganisms use a series of enzymes, called cascade enzymes, to break down pollutants step by step into less toxic end products. However, this process is slow and often allows toxic intermediates to accumulate. The goal of this CAREER project is to make biodegradation more efficient. The project will develop a new biotechnology, called protein nano-compartment (PNC)-based cargo encapsulation. Cascade enzymes will be encapsulated within PNCs, which will enable the enzymes to degrade pollutants and intermediates at similar rates. Toxic intermediates will not accumulate in the environment. The research will be integrated with education of students from middle schools and colleges. Successful completion of this project will create a more efficient, robust, and faster environmental remediation technology to protect human and environmental health. This CAREER project plans to apply PNC-based enzyme co-localization to accelerate biodegradation efficiency in removing organic water contaminants. The central hypothesis is that attaching enzymes with affinity tags of varying molecular properties will allow their tunable co-localization within PNCs, thereby enabling optimization and enhancement of the kinetics and stability of enzyme cascades for contaminant degradation. The study will integrate techniques in biodegradation, synthetic biology, an