Surface Effect of Assembling Enzyme and Modulation of Surface Enzyme Activity with Electric Potential Stress

Abstract

The fluorescent marker of rhodamine B amine is successfully used to evaluate the immobilization capability onto silicon-based patterns fabricated by semiconductor manufacturing. Only the silicon dioxide surface, by means of fluorescent observation, can immobilize the rhodamine molecule by the sequential linkage of (3-aminopropyl) triethoxysilane (APTES) and glutaraldehyde. The phenol sulfotransferase enzyme is also successfully immobilized onto the silicon dioxide surface by the linking molecules of APTES and sulfosuccinimidyl 4-(N-maleimidomethyl)-cyclohezane-1-carboxylate in the home-made apparatus. The enzyme activity of the sulfotransferase is determined from the absorbance of 4-nitrophenol at 400 nm wavelength. The surface immobilized enzyme remains its activity for catalytic reaction at least 120-min duration. The surface saturation effect on the activity of immobilized enzyme is explained and ascribed to the surface diffusion effect of electric double layers. We can success control the surface immobilized enzyme by electric potential stress. The activity of enzyme is reduced under negative potential, while is enhanced under positive potential. The electric potential can induce the enzyme structure variation and modulate the enzyme activity due to the electrostatic effect.