Engineered Oxalate Decarboxylase Boosts Activity and Stability for Biological Applications Dindo, Mirco Conter, Carolina Uechi, Gen-Ichiro Pampalone, Gioena Ruta, Luana Pey Rodríguez, Ángel Luis Rossi, Luigia Laurino, Paola Magnani, Mauro Cellini, Barbara Oxalate decarboxylase (OxDC) from Bacillus subtilis is a Mn-dependent hexameric enzyme that converts oxalate to carbon dioxide and formate. Recently, OxDC has attracted the interest of the scientific community due to its biotechnological and medical applications for the treatment of hyperoxaluria, a group of pathologic conditions associated with excessive oxalate urinary excretion caused by either increased endogenous production or increased exogenous absorption. The fact that OxDC displays optimum pH in the acidic range represents a big limitation for most biotechnological applications involving processes occurring at neutral pH, where the activity and stability of the enzyme are remarkably reduced. Here, through bioinformatics-guided protein engineering followed by combinatorial mutagenesis and analyses of activity and thermal stability, we identified a double mutant of OxDC endowed with enhanced catalytic efficiency and stability under physiological conditions. The obtained engineered form of OxDC offers a potential tool for improved intestinal oxalate degradation in hyperoxaluria patients. 2025-04-04T09:35:17Z 2025-04-04T09:35:17Z 2025-03-23 journal article ACS Omega 2025, 10, 12, 12375–12384 [https://doi.org/10.1021/acsomega.4c11434] https://hdl.handle.net/10481/103445 eng http://creativecommons.org/licenses/by/4.0/ open access Atribución 4.0 Internacional American Chemical Society