A team of scientists from the Pacific Northwest National Laboratory (PNNL) and the Environmental Molecular Sciences Laboratory (EMSL), a facility supported by the U.S. Department of Energy, has created a new tool to identify enzymes in soil bacteria that break down complex molecules. These enzymes, called amide hydrolases, help break down amide bonds, which are found in many biological substances. The researchers focused on bacteria that naturally break down chitin, a tough, natural polymer found in the shells of insects, crustaceans, and the cell walls of fungi. The method uses specially designed synthetic compounds that glow when exposed to certain enzymes. These compounds are broken down by microbial enzymes, creating probes that reveal the presence of specific enzymes. The process involves two main steps: first, using these glowing compounds to detect enzyme activity in real time, and then converting those signals into tags that help identify the specific proteins responsible. This approach helps connect the genetic code of an organism to the actual function of its proteins, which traditional methods often miss. By using a large library of chemical probes, the researchers were able to identify amide hydrolases in complex communities of soil bacteria. To further pinpoint the responsible proteins, the team added a chemical tag to the most active compound, which allowed them to isolate and identify the proteins using mass spectrometry, a technique that determines the molecular structure of substances. The study shows how a flexible library of glowing compounds can be used to map out enzymes in their natural environments, even when those enzymes have not been previously identified or studied. This method could lead to new discoveries in biotechnology, such as finding bacteria that can break down synthetic materials like nylon in an environmentally friendly way. The research was published in Chemical Communications, a scientific journal that covers advances in chemistry.