Storage proteins from
Sphenostylis stenocarpa
and
Phaseolus lunatus
were fractionated, and their
in vitro
bioactivities were investigated. Albumin, globulin, prolamin, and glutelin constituents of the respective seeds were successively fractionated using the modified Osborne method. Phenylmethylsulfonyl fluoride (1 mM) was used as a protease inhibitor. The antioxidant, anti-inflammatory, and acetylcholinesterase-inhibitory activities of the protein fractions were evaluated using different appropriate techniques. Globulin was the predominant fraction, with a yield of 43.21±0.01% and 48.19±0.03% for
S. stenocarpa
and
P. lunatus
, respectively, whereas prolamin was not detected in both seeds. The protein fraction markedly scavenges hydroxyl radicals, nitric oxide radicals, and 2,2-diphenyl-1-picryldydrazyl radicals with concomitant high free radical-reducing power. Albumin and globulin fractions elicited the highest acetylcholinesterase-inhibitory potential of 48.75% and 49.75%, respectively, indicating their great application potential in managing neurodegenerative diseases. In this study, the albumin, globulin, and glutelin fractions of these underutilized legumes showed great analeptic bioactivities, which could be utilized as health-promoting dietary supplements/products.