Use of immobilized amyloglucosidase as chiral selector in chromatography. Immobilization and performance in liquid chromatography

The enzyme amyloglucosidase was immobilized on oxidized DIOL silica and use d to separate enantiomers of amino alcohols. The influence of pore size On enantioselectivity was studied and an optimum in the separation factors was found using 500 A DIOL silica as the starting material. About twice the am ount of the protein could be immobilized on the 500 Angstrom DIOL silica ma terial as on the 300 and 1000 Angstrom materials. The immobilization proced ure was easy to reproduce and no significant difference in the chromatograp hic behavior was observed between two amyloglucosidase columns produced in- house. The effect of solute structure on enantioselective retention was stu died using a set of 10 closely related amino alcohols. High separation fact ors (alpha > 2) were obtained and the efficiency of the amyloglucosidase co lumns was greater than 25 000 plates/m based on the last eluted enantiomer.