Pseudomonas species is known to cause multiple nosocomial infections in patients and results in high morbidity and mortality rates (10%). The greatest obstacle in treating patients infected with the Pseudomonas species is the widespread emergence of antibiotic resistance. Hence, there is an urgent need to develop new compounds which can be effective against Pseudomonas species and possibly remain tolerant to drug resistance. The enzyme glutamate racemase plays an important role in cell wall synthesis of bacteria and as a rate limiting step, thus it is an excellent target for the designing of new class of antibacterial agents. The objective of this study is to investigate the variations in sequences of glutamate racemase, a potential drug target across the all 31 species of Pseudomonas. Sequence variability and conservation for functional motif identification is helpful for identifying evolutionarily important residues with functional significance; subsequently these results of variable sites were supported by entropy profile obtained from protein variability server using Shannon entropy. Phylogenetic profile among the different Pseudomonas sp. having fully/highly conserved residues was observed, suggesting possible functional similarities between them. The variation analysis in conserved and non-conserved region of the sequence can be used to predict the binding site for target specific drug discovery.