Lectins are carbohydrate-binding proteins that play vital roles in many biological processes. In this study, lectins from three Egyptian cultivars (fava bean, lentil, and pea) were isolated by precipitation with different concentrations of ammonium sulfate. The purification process was performed by affinity chromatography using mannose agarose. The highest concentration of purified lectins (1.48 mg/g) was recorded in pea at 90% saturation. SDS-PAGE of the purified lectins revealed bands of low molecular weights (14 to 18 kDa). The complete amino acid sequences of purified lectins were assessed using mass spectrometry (MS), which indicated the presence of the peptides favin, p54, and psl in fava bean, lentil, and pea, respectively. The lectins showed antimicrobial activity. The highest inhibition zone (35 mm) was measured with lectin purified from lentil against Staphylococcus aureus ATCC 6538, followed by pea lectin (33.4 mm) against Pseudomonas aeruginosa ATCC 10145. To the best of our knowledge, the legume lectins in this study are the first lectins to exhibit antifungal activity against Candida albicans, with the maximum inhibition zone (25.1 mm) observed with purified lectins of fava bean. Additionally, the first scanning electron microscope (SEM) images showing agglutination and clumping of microbial cells exposed to tested lectins are provided. These findings proved that Egyptian legume lectins are distinct from other lectins reported in previous studies and demonstrated their potential as antimicrobial agents against human pathogenic microorganisms.