The aim of this work was to study the processes taking place during the enzymatic hydrolysis of seed globulins isolated from narrow-leaf (var. Zeus and Bojar) and yellow (var. Lord and Parys) lupin seeds species cultivated in Poland. In lupin seed globulins hydrolysis studies, there were used enzymes typical for the human gastrointestinal tract, such as: pepsin, pancreatin, trypsin and chymotrypsin. The obtained hydrolysates were assessed based on the results of electrophoretic, immunoblotting, as well as chromatographic separations. The evaluation of lupin seed globulins digestion susceptibility was supported by the bioinformatics analyses. Analysis of hydrolysates showed that the proteins present in globulins are completely hydrolysed by: pepsin, double digestion model (pepsin followed by pancreatin) or chymotrypsin. The high specificity of pancreatin and trypsin action results in limited lupin globulins hydrolysis. Protein fraction resistant to the action of these enzymes was γ-conglutin which also retains its antigenic properties. Its insensitivity to the hydrolysis might be associated with the formation of complexes with flavonoids which were released from other protein connections during digestion, as well as with relatively low number of cleavage sites for trypsin. The analysis of the three-dimensional structure of γ-conglutin enabled a very accurate description of the amino acid residues localisation, at which trypsin hydrolysis should occur.