A stepwise approach to the understanding of extracellular enzyme activity in soil. I. Effect of electrostatic interactions on the conformation of a beta-D-glucosidase adsorbed on different mineral surfaces.
The enzymatic activity of sweet almond beta-D-glucosidase adsorbed on various mineral surfaces was studied. Our aim was to elucidate the mechanism responsible for the observed changes in catalytic activity. The results of the investigation are discussed with reference to the hypotheses generally proposed to explain the well-documented shift in optimal pH of the activity of adsorbed enzymes. By separate determinations of enzymatic activity in a mineral suspension and of its supernatant solution, and comparison with a control without mineral added, we obtained accurate measurements of the catalytic activity of the adsorbed enzyme alone. Different pH profiles of activity profiles were found when the enzyme was adsorbed onto montmorillonite, kaolinite and goethite. The activity profiles, were also found to vary with ionic strength, the pH at which enzyme adsorbed onto the mineral surface, and in the case of goethite, on the nature of the anions in the buffer. Our observations cannot be adequately explained by assuming a more acidic microenvironment at the mineral surface. We postulate that on some mineral surfaces a conformational change is induced in the adsorbed protein, which reduces its catalytic activity. We contend that such conformational changes are due primarily to electrostatic forces.
D000327:Adsorption; D001546:Bentonite; D002021:Buffers; D005959:Glucosidases / Q000302:isolation & purification*; D006863:Hydrogen-Ion Concentration; D008903:Minerals / Q000032:analysis*; D011487:Protein Conformation; D012987:Soil / Q000032:analysis*; D013379:Substrate Specificity; D013499:Surface Properties; D001617:beta-Glucosidase / Q000302:isolation & purification*
D002021:Buffers; D008903:Minerals; D012987:Soil; D001546:Bentonite; D005959:Glucosidases; D001617:beta-Glucosidase