Cross-linked glucose isomerase crystals as a liquid chromatographic separation material

Cross-linked crystals of glucose isomerase (CLGI) were characterized as a l iquid chromatographic separation material. The experiments were done with c rystals having an average diameter of 83 mu m. Porosity (epsilon(p)) and po re size distribution of the CLGI crystals were measured with size exclusion chromatography using D2O and polyethylene glycols as probes. CLGI material was capable of separating (1000 g/mol polyethylene glycols. Fifty two perc ent (epsilon(p) = 0.47) of the total crystal volume was in pores. Pore size measurement showed that CLGI crystals were microporous material, having an average apparent pore diameter of 29 +/- 0.08 Angstrom. CLGI material sepa rated n-alcohols C-1 to C-8 based on the hydrophobic interaction between th e protein material and the carbon chain of the alcohols. Height equivalent to a theoretical plate (HETP, in millimeters) ranged from 1.6 to 0.89 for t he C-1 to C-7 n-alcohol series. Despite the large crystal size, CLGI as a c hirally active phase effectively separated D- and L-arabitol (R-s = 0.58) a nd showed potential for chiral separation of amino acids. (C) 2000 Elsevier Science Inc. All rights reserved.