Stabilization of dimeric β-glucosidase from Aspergillus niger via glutaraldehyde immobilization under different conditions.

Publication date: Available online 22 December 2017 Source:Enzyme and Microbial Technology Author(s): Perla Guadalupe Vazquez-Ortega, Maria Teresa Alcaraz-Fructuoso, Juan A. Rojas-Contreras, Javier López-Miranda, Roberto Fernandez-Lafuente The dimeric enzyme β-glucosidase from Aspergillus niger has been immobilized on different amino-agarose beads at pH 5 and 7, exploiting the versatility of glutaraldehyde. The stability of the free enzyme depended on enzyme concentration. Immobilization via ion exchange improved enzyme stability/activity, depending on the immobilization pH. However, the enzyme was desorbed in 75 mM NaCl at pH 7 and some stability/enzyme concentration dependence still existed. Treatment: of these biocatalysts with glutaraldehyde increased enzyme stability (e.g. at pH 5, after incubation under conditions where the enzyme just ionically exchanged was fully inactivated, the activity of the glutaraldehyde treated enzyme remained unaltered). Immobilization on glutaraldehyde pre-activated supports yielded a higher increase in enzyme activity, but the stabilization was lower. While when measuring the enzyme activity at pH 4 there were no changes after immobilization, all immobilized enzymes were more active than the free enzyme at pH 6 and 7 (2–3 times). The Ki/Km ratio did not significantly decrease in any immobilized biocatalysts, and in some cases it worsened in a significant way (by a 9 fold factor using preactivated supports). The new biocatalysts ...
Source: Enzyme and Microbial Technology - Category: Biotechnology Source Type: research