The effect of Osmolytes (sucrose and glucose) on bovine intestine Alkaline Phosphatase activity

Introduction: One of the most important purposes of enzyme engineering is to increase the thermal and kinetic stability of enzymes. One of the best methods to increase the stability and activity of enzymes is modification of the solvent environment. Materials and methods: The purpose of the present study was to try for increasing the activity of bovine intestine alkaline phosphatase (BIALP) by modification of the solvent environment. So, we study how sucrose and glucose could effect on the conformation, and the function of BIALP. The influence of this modifier explored using fluorescence spectroscopy and kinetics studies. Results: We have determined the thermodynamic parameters, kinetic constant enthalpy change (ΔH°), entropy change (ΔS°) and Gibbs free energy change (ΔG°) to understand BIALP stability and activity. Fluorescence spectroscopy revealing the static mode of the quenching. The number of binding sites was 1. Fluorescence spectroscopy revealed that the hydrogen bonding and Vander Waals forces played a major role in stabilizing the complex. Activity of this complex was enhanced due to the higher H-bond formation and the lower surface hydrophobicity after sucrose and glucose modification. Conclusion: Sucrose and glucose acted as enhancers for BIALP activity. The result reveals the ability of sucrose and glucose to protect the native structural conformation of BIALP. These results explicitly explain that stabilizing sucrose and glucose is preferentially excluded from the surface of BIALP, since water has a higher tendency toward favorable interactions with functional groups of BIALP than with sucrose and glucose

کلیدواژه ها:

BIALP ، Sucrose ، Glucose ، Fluorescence spectrophotometry ، Entropy change ، Enthalpy change