Galactose Oxidase Immobilized on Silica in an Analytical Determination of Galactose-containing Carbohydrates

Abstract

Galactose oxidase from Fusarium graminearum IMV-1060 adsorbed on, and covalently bound to, silica carriers has been used for analytical determinations of D-galactose and galactose-containing sugars. Using a flowing oxygen electrode of the Clark-type, sensor system for enzymatic analysis of water solutions of galactose-containing carbohydrates was made. Measurements were taken both in the pulse and continuous modes of a substrate flowing through a column with an immobilized biocatalyst. The linear measurement ranges for galactose-containing carbohydrates concentrations were determined.

Practical Synthesis of (20S)‐10‐(3‐Aminopropyloxy)‐7‐ethylcamptothecin, a Water‐Soluble Analogue of Camptothecin

“…Similarly, Kondaka et al. have shown that GAOx is strongly adsorbed on silica surface, which is also negatively charged at pH 7, with an enhanced enzyme activity compared to the free enzyme 18. Laponite was therefore selected as immobilization matrix for GAOx…”

Selective Oxidation of 5‐Hydroxymethylfurfural to 2,5‐Diformylfuran in Biphasic Media using Immobilized Galactose Oxidase: Proof of Concept and Limitations

“…[22] In another approach, GalOx was immobilized onto silica beads via adsorption and covalent attachment. [23] GalOx immobilized on mesoporous silica particles displayed increased thermal resistance to 60 °C. [24] Immobilization of GalOx on chemically-modified porous silica beads enabled its use in a packed bed reactor.…”

Probing structural and catalytic characteristics of galactose oxidase confined in nanoscale chemical environments

“…Moreover, the exact role of the semi-reduced state is still not fully understood, although it is likely to be physiologically relevant. 5 (c) The enzyme has an unusually high isoelectric point (pI) of 12,6 leading to a signicantly positively charged state in almost all buffers commonly used.…”

“…9 To construct GAOX based biosensors, immobilization of this enzyme on different supports including macroporous silica has been attempted. [10][11][12] Immobilization of enzymes on a support material is essential in many practical applications such as industrial biocatalysis, biosensors, and biofuel cells. Study on the immobilization of enzymes started almost a century ago 13 and has since then been extensively conducted.…”

“…In an early attempt, macroporous silica was used to immobilize GAOX for analytical detection of galactosecontaining carbohydrate. 12 Due to incompatible pore sizes (too large, 40-60 nm), 47 however, enzyme leaching was a serious problem and the experimental results were inconclusive. To the best of our knowledge, immobilization of galactose oxidase on mesoporous silica has not been reported before.…”