Objective: The current investigation involves the purification, characterization of the lectin from the leaves of Pimenta dioica (L.) Merr. (Myrtaceae) a medicinal plant, and its application in bacterial typing.Methods: A lectin was purified from the leaves by cation exchange chromatography. SDS PAGE revealed the molecular weight of the purified lectin. Biochemical characterization was carried out by performing various tests. Hemagglutination inhibition was conducted to detect the sugar specificity. Additionally, bacterial agglutination was performed to predict whether the purified lectin was able to agglutinate the bacterial strains.Results: SDS PAGE analysis revealed the lectin to be a tetramer in the range of 43-66 kDa. The purified lectin agglutinated human, avian, and mouse erythrocytes, and was inhibited by 125 mmol of mannose and xylose. The lectin was stable at 0-60 ° C for 30 min and was unaffected by either 2-Mercaptoethanol (2-ME) or Dithiothreitol (DTT) (50-250µM). A pH of 6.0–8.0 was found optimum for its activity and was nearly independent of metal ions. The purified lectin contained about 20% carbohydrate as estimated by Anthrone method. Purified lectin agglutinated the Gram-negative Escherichia coli and Proteus vulgaris.Conclusion: The isolated lectin was found to possess significant hemagglutinating activity. Due to its ability to agglutinate Gram negative bacteria such as Escherichia coli and Proteus vulgaris, it could be used for bacterial typing and for the design of bacterial filters.