Introdutction:The present study was designed to screen out the different proteins from Prosopis cineraria (Leaves), known as Shammi (Family Fabaceae) and characterize them by IR, NMR and mass spectroscopy and evaluation of their antimicrobial potential against various microbial species. Mehtods: Different extracts of leaf were treated with different reagents for the presence and absence of metabolites. Antimicrobial effect of the proteins were explored against Staphylococcus aureus, Pseudomonas aeruginosa and Aspergillus tubingensis. The disc diffusion method was used for deter mination of antimicrobial activity. Analytical analysis included IR, NMR and Mass spectroscopy. Reults: Phytochemical analysis of different extracts showed the presence of different primary and secondary metabolites The range of zone of inhibition were found to be from 1.4±0.01mm to 14.2±0.18 mm. The maximum antimicrobial activity was found for 50 KDa protein against Pseudomonas aeruginosa.