Enolase (EC 188.8.131.52, 2-phosphoglycerate dehydratase) catalysis the ninth and penultimate step of glycolysis which is the conversion of 2-phosphoglycerate to phosphoenolpyruvate. Toxoplasma gondii encodes two isoforms of such glycolytic enzymes, enolase 1 and enolase 2 that are expressed exclusively in bradyzoites and tachyzoites, respectively. The brain cysts specific TgEnolase 1 of the slow growing bradyzoites has been implicated as an attributed target in drug development for Toxoplasmosis. Despite its sequence similarity to other enolases, the TgEnolase 1 dimmer structure exhibits distinct surface properties. The EWGWS pentapeptide insertion and insertions in catalytic loops (loop 3) separate TgEnolase from the host human neuron specific enolase (hENO2, PDB code 1TE6). The C’-terminal KSGSPCRSERLCKYNQLMR sequences of each TgEnolase 1 monomers create a positively charged area that may correspond to the unique DNA binding function of TgEnolase1. These features indicate the role of TgEnolase 1 in the important pathogen T. gondii.