In this work, we studied the immobilization of enolase, a glycolytic enzyme, onto silicon wafers, amino-terminated surfaces, polystyrene (PS) and poly(methyl methacrylate) (PMMA) by means of in situ ellipsometry and contact angle measurements. The adsorption kinetics of enolase on the substrates revealed a continuous increase of the adsorbed amount as a function of the time. This effect might be attributed to the cooperativity phenomena, which is common to enzymes and proteins. The mean diffusion coefficient (D) value determined for enolase amounted to 10(-7) and 10(-9) cm(2)/s, in NaCl 10(-3) mol/L and Tris-HCl buffer, respectively, The adsorption isotherms revealed a preferential adsorption of enolase onto amino-terminated surfaces and PMMA films. The enolase biofilms presented hydrophobic character (theta(a) = 74degrees +/- 3degrees), indicating the exposure of the enolase hydrophobic segments to the air.