DL-3-Chloroalanine is primarily used in biochemical research as an inhibitor of pyridoxal phosphate-dependent enzymes, particularly alanine racemase and other aminotransferases. It acts as a substrate analog, interfering with amino acid metabolism in microbial and mammalian systems. This compound is valuable in studies exploring bacterial cell wall biosynthesis, as it can disrupt peptidoglycan formation by inhibiting the production of D-alanine, a key component. Due to this mechanism, it has been investigated for its potential antimicrobial properties and as a lead compound in developing new antibiotics. Additionally, DL-3-Chloroalanine is employed in enzyme kinetics studies to understand reaction mechanisms involving amino acid racemization and transamination. Its ability to cross cell membranes makes it useful in cellular and metabolic labeling experiments.