Edible legume sprouts have been proposed as a promising plant-based source of the enzyme diamine oxidase (DAO), which plays a key role in degrading histamine at an intestinal level and preventing the development of histamine intolerance symptoms. However, the temperature and humidity conditions required for seed germination can also favor the rapid growth of yeast and mold, potentially compromising sprout yield and quality. The aim of this study was to evaluate the influence of different seed disinfection treatments on both the germination rate and DAO enzymatic activity in sprouts of four Leguminosae species. Seed disinfection with 70% ethanol for either 5 or 15 min slightly increased the germination rates of chickpea and soybean sprouts without affecting DAO activity, regardless of treatment duration. However, in lentil and green pea sprouts, ethanol disinfection caused a statistically significant reduction in histamine-degrading capacity. In contrast, treating seeds with sodium hypochlorite for 15 min increased germination rates by up to 14% and preserved DAO activity in all legume sprouts tested. These results indicate that incorporating a seed disinfection step during legume sprouting may affect both the DAO enzymatic activity and germination rate.