Plant Protease inhibitors (PIs) are widely studied defense proteins of the Plants. In seed and other tissues of the plant, protease inhibitors are found which represents up to 10% of the total protein (Casaretto and Corcuera 1995). PI inhibits tryptic and chymotryptic gut proteinases which affect protein digestion in the insects. This leads to the overproduction of gut proteases to compensate for the inhibited activity leading to deficiency of essential amino acids. This creates physiological stress and affects growth of the insect pests. The different Vigna Accessions as Vigna hainiana, Vigna aconitifolia and Vigna sublobata found to have potent Helicoverpa armigera insect gut proteolytic activity. Hence, efforts have been made to isolate and purify the protease inhibitors from these 03 Vigna Species. Ammonium Sulphate salt precipitation, Ion Exchange Chromatography and PAGE techniques have been used for the Purification of Protease Inhibitors. The single protein purified bands of different Vigna such as V. hainiana, V. aconitifolia and V. sublobata showed molecular weight of 8.18 KDa, 10.66 KDa and 10.26 KDa respectively when compared with standard molecular weight markers.