Kinetics and Spectrofluorimetric studies of alkaline phosphatase in presence of ribose

Alkaline phosphatase from bovine intestinal mucosa is a homodimeric metalloenzyme that hydrolyze non-specifically phosphate monoesters at alkaline pH to produce inorganic phosphate and alcohol. This enzyme has many application in the diagnosis, immunology and molecular cell biology. Hence, change in activity and a main goal. The results of kinetic studies indicated this sugar conformation of BIALP is important and follow as effected on enzyme activity, increased Km and decreased Vmax so ribose acted as a mix inhibitor and bind at a site that distinct from the substrate active site. Also obtained results from fluorescence studies showed in presence of ribose fluorescence emission reduced and occurred in high wavelengths (redshift) so ribose changed plot microenvironment of BIALP’s Tryptophan residues and acted as a quencher. The slope of Stern_Volmer increased with increasing temperature and fluorescence quenching constants (kq) at two temperature were much lower than maximal dynamic quenching constant (2×10+10 M-1S-1) so dynamic quenching were occurred.

Keywords:

(BIALP) , ribose , Kinetics studies , Fluorescence studies Alkaline phosphatase from bovine intestinal mucosa