Temperature and pH effects with immobilized electric eel acetylcholinesterase.
Kinetic studies were made with 2 forms of immobilized acetylcholinesterase: enzyme trapped in polyacrylamide gel which was cut into slices; and enzyme attached to the inner surface of nylon tubing. Rates were measured at substrate concentrations which were low and high with reference to the Michaelis constant, and over the temperature range 16-40 degrees C. Low activation energies (1.7-2.7 kcal mol-1) were obtained at low substrate concentrations, indicating diffusion control. At high substrate concentrations the Arrhenius plots were non-linear and the activation energies substantially higher, and there is less diffusion control. With enzyme-polyacrylamide slices, there was a continuous increase in rate with increasing pH, in contrast to the bell-shaped behavior with free enzyme. A theoretical treatment suggests that this is due to the lowering of local pH as a result of the acid released in the hydrolysis.
Temperature and pH effects with immobilized electric eel acetylcholinesterase. Kinetic studies were made with 2 forms of immobilized acetylcholinesterase: enzyme trapped in polyacrylamide gel which was cut into slices; and enzyme attached to the inner surface of nylon tubing. Rates were measured at substrate concentrations which were low and high with reference to the Michaelis constant, and over the temperature range 16-40 degrees C. Low activation energies (1.7-2.7 kcal mol-1) were obtained at low substrate concentrations, indicating diffusion control. At high substrate concentrations the Arrhenius plots were non-linear and the activation energies substantially higher, and there is less diffusion control. With enzyme-polyacrylamide slices, there was a continuous increase in rate with increasing pH, in contrast to the bell-shaped behavior with free enzyme. A theoretical treatment suggests that this is due to the lowering of local pH as a result of the acid released in the hydrolysis.