A process for the recovery and immobilization of starch phosphorylase from starch-based industrial wastewater

摘要

A process for recovery of starch phosphorylase, an important enzyme in the production of glucose 1-phosphate from the wastewater of potato-starch factories, has been developed. Of the 680 units of enzyme present in 1.2 litres of wastewater, 383 units were recovered with a 2.72-fold enrichment in specific activity. Two forms, starch phosphorylase Ⅰ and Ⅱ, obtained by ion-exchange cnromatography, were purified to a specific activity of 0.76 and 0.82 unit/mg of protein respectively, which remained essentially unchanged even after immobilization on ground eggshell. However, the thermal stability of both the immobilized enzymes was dramatically increased. The eggshell-bound enzymes were packed in a water-jacketed glass column and fed with a continuous supply of soluble starch and phosphate buffer to yield glucose 1-phosphate, a cytostatic compound used in the therapy of cardiopathy, which was isolated by DEAE-cellulose ion-exchange chromatography and ethanol precipitation to 96% purity.