A protease inhibitor (PI) was isolated and purified from halo-alkaliphilic Streptomyces spp. VL J2. SDS-PAGE of purified PI revealed it to be a heterodimer of two unidentical subunits of 27.5 and 11.08 kDa, which corroborates well with intact molecular mass of 38.5 kDa obtained by GPC and MALDI -TOF. Inhibitory activity was confirmed by activity staining and reverse zymogram studies and the inhibitor was found to retain activity at <50 degrees C and pH 2-9.5. It showed presence of two isoforms with isoelectric point of 5.5 and 5.7. The inhibition of trypsin and chymotrypsin indicated it to be a serine protease type belonging to serpin family. The stoichiometry of trypsin-inhibitor interaction was 1:2. Modification of amino acid showed presence of arginine and free sulfhydryl group at active site. Kinetic studies revealed the non-competitive type of inhibition of trypsin with low Ki value (9.4 x 10(-9) M). Activity against H. armigera showed significant decline and delay in larval (51%), pupal weights and periods, respectively, prominent physical abnormalities and reduced nutritional indices as a function of treatment. The results suggest that the purified PI has promising pesticidal activity and could serve as a potential candidate gene for transgenic plant research. (C) 2016 Elsevier Ltd. All rights reserved.