The role of hydration in enzyme activity and stability: 1. Water adsorption by alcohol dehydrogenase in a continuous gas phase reactor.
The adsorption of water by alcohol dehydrogenase from baker's yeast (YADH) has been measured in a continuous-flow gas reactor at varying temperatures. Adsorption isotherms in the presence of gaseous organic substrates are compared to those from organic-free gas mixtures. Almost no effect of the hydrophobic molecule on total water adsorption was observed. A rarely mentioned multilayer isotherm model from the 1930s, the Huttig's isotherm, has been found to fit the experimental data with extremely good accuracy. The model enables the calculation of both the heat of adsorption of water to the enzyme and the total amount of water necessary for monolayer coverage. The heat of adsorption of water in the first layer is approximately -16 kcal/mol. This tight binding of water, which is much higher than the heat of condensation of pure water, helps to explain the kinetic properties of YADH-catalyzed reactions on vapor phase substrates. While the monolayer coverage is temperature independent, the enzyme demonstrates hysteresis when transitioning between adsorption and desorption. The hysteresis observed in water sorption studies may also explain previously reported properties of the enzyme. (c) 1996 John Wiley & Sons, Inc.
Yang
F
F
Department of Chemical Engineering & Center for Biotechnology and Bioengineering, University of Pittsburgh, Pittsburgh, Pennsylvania, 15261.
Russell
A J
AJ