This article has been reviewed according to Science X's editorial process and policies. Editors have highlighted the following attributes while ensuring the content's credibility: Proteins are vital to living systems. They build and repair tissues, act as enzymes to speed chemical reactions, regulate hormones and transport nutrients—all of which are important for developing next-generation bioproducts and advancing bioengineering and biotechnology.
Determining protein function is difficult using conventional scientific approaches because a specific protein often has more than one function. Complementary methods are needed to understand proteins' functional contributions to phenotype—how genes are expressed as observable traits within an environment. Chemical probes are uniquely suited to bridge the gap between knowing the sequence of genes for a particular protein and what the protein does because they reveal enzyme activity directly, allowing researchers to connect predicted genes with a real biochemical function.
A fluorogenic substrate library was developed by a team of researchers from Pacific Northwest National Laboratory (PNNL) and the Environmental Molecular Sciences Laboratory (EMSL), a Department of Energy (DOE) Office of Science user facility. The library was used to visualize the activity of amide hydrolase (an enzyme that catalyzes the breakdown of amide bonds found in many biological molecules) from soil-derived bacteria that naturally degrade chitin (a tough natural polymer found in fungal cell walls, insect exoskeletons and crustacean shells). Synthetic fluorogenic compounds cleaved by microbial enzymes were converted into activity-based probes that enabled the identification of hydrolases with broad substrate tolerance.
The development of this approach bridges the gap between genomic sequence information and the biological function of specific proteins by identifying active enzymes that traditional analysis methods often overlook. By deploying a high-throughput library of chemical probes, the study successfully identified amide hydrolases in complex soil bacteria. The core innovation lies in the two-step identification-and-capture workflow: using fluorogenic substrates to detect enzymatic activity in real time and converting those detections into tags for protein identification.
This method marks a significant advancement for profiling microbial proteins with unknown functions. Potential applications include environmental microbiology, biomanufacturing and medicine. This research was published in ChemComm.
In this study, a team of researchers from PNNL and EMSL developed a fluorescence chemical probe library to target amide hydrolase. They tested the library using soil-derived chitin-degrading bacteria to detect amide hydrolase activity. Researchers added a chemical tag to the highly active compound that enabled the enrichment of proteins.
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