Carboxypeptidase G, derived from Pseudomonas sp., is primarily utilized in medical and biotechnological applications. It plays a critical role in cancer therapy, specifically in the treatment of methotrexate toxicity. Methotrexate, a chemotherapy drug, can accumulate to toxic levels in patients with impaired renal function. This enzyme effectively cleaves methotrexate into non-toxic metabolites, aiding in its rapid elimination from the body. Additionally, it is employed in enzymatic assays and research studies to investigate peptide and protein structures due to its specificity in cleaving C-terminal glutamate residues. Its application extends to the development of targeted drug delivery systems, where it is used to activate prodrugs selectively at tumor sites, enhancing therapeutic efficacy while minimizing side effects.