Tilapia mossambica is a eurythermal tropical fish. We studied the effect of temperature on the kinetics of brain Acetylcholinesterase (AChE) during adaptation to sublethal temperatures by acclimating the fish to 37 °C, and controls to 25 °C. Electrophoresis showed the presence of two AChE bands that did not change in position or intensity with acclimation period or temperature. The apparent Km was 0.23 ± 0.01 mM ATChI and remained relatively constant over the in vitro assay temperature range 10 °C to 40 °C. Biochemical characterization suggested that the enzyme is a 'eurytolerant protein' in its stability of kinetic and thermal properties over a wide temperature range. Thermal stability and arrhenius plots suggested that the AChE was made up of two forms that differed in their thermal properties.The two molecular forms of acetylcholinesterase were purified from the brain of T. mossambica. Molecular weight studies revealed that the two forms were size isomers: a monomer of 59 KDa and a tetramer of 244 KDa. They differed in their Kms, thermal stabilities and energies of activation. We suggest that biochemical adaptation to temperature in the brain acetylcholinerase system of the fish Tilapia mossambica is based on the aggregation-dissociation of these size isomers.