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Biostimulants are sustainable inputs that can be used to reduce chemical fertilizers application, improving nutrient absorption, production and quality of crops. Protein Hydrolysates (PH) are a category of biostimulants used to counteract reductions in production in non-optimal nitrogen (N) conditions. On this basis, a comparison was made between a new PH derived from the Malvaceae botanical family and a commercial PH derived from the Fabaceae botanical family on Lactuca sativa L. var. capitata cv. “Maravilla De Verano Canasta” grown in a cold greenhouse (from 2 October for 42 days) at the Department of Agriculture (Portici, NA), in a container on an inert substrate (quartz sand) with optimal concentrations (8 mM NO3-) and N deficiency (1 mM NO3-) in the nutrient solution. Both PHs improved biometric parameters including production, although to a lesser extent under nitrogen deficiency conditions. PH derived from legumes recorded higher values of Fv/Fm, lutein and β-carotene as well as higher levels of catalase and phenolic acids totals. In contrast, Ph derived from Malvaceae did not influence the content of total phenolic acids but increased the leaf concentration of Ca, Mg and catalase, reducing H2O2. Among the different organic compounds synthesized by plants, phenylpropanoids were mainly accumulated in plants subjected to nutritional stress. In conclusion, the application of IP, although inducing a general improvement in nutrient absorption in optimal conditions, cannot compensate for the negative effects of a severe N deficiency.