SummaryThe objective was to evaluate the in vitro inhibition of enzymes related to neurodegeneration (prolyl endopeptidase, PEP; acetylcholinesterase, AChE; butyrylcholinesterase, BChE; lipoxygenase, LOX) and the antioxidant activity from fermented milks (FM) with Lactococcus and Lactobacillus spp. All FM presented antioxidant activity (ABTS and ORAC); however, FM with Lactococcus NRRL B‐50571 (FM‐571) presented higher (P < 0.05) oxidative haemolysis inhibition. Additionally, FM‐571 presented the lowest IC₅₀ for LOX (1.78 mg/mL), PEP (3.16 mg/mL), AChE (3.22 mg/mL) and BChE (2.14 mg/mL). Thus, FM‐571 was selected for in silico analysis for the prediction of binding sites of peptides present in this FM with enzymes active sites. Altogether, thirty peptides showed (P < 0.01) high binding potential to AChE, seventeen to BChE and LOX and sixteen to PEP. Thus, FM‐571 shows potential for the development of dairy products with potential neuroprotective effects.