Immobilization of bile acid enzymes onto activated amino-ethyl cellulose and polyvinyl alcohol membranes

Abstract

Immobilization of bile acid enzymes named, 3-alpha hydroxysteroid dehydrogenase (3-α HSD) and lipoyl dehydrogenase (Diaphorase) enzymes was done on to activated polymeric membranes like amino-ethyl cellulose and Polyvinyl Alcohol. Immobilization of bile acid enzymes was done onto activated amino-ethyl cellulose by covalent coupling by using NaOH and glutaraldehyde. And, Immobilization of bile acid enzymes was done onto activated polyvinyl alcohol by using terephthaladehyde and 2-(m-aminophenyl)-1-3-dioxolane. Percent of enzyme coupled was found 65% and 60% for enzymes coupled onto activated amino-ethyl cellulose and activated polyvinyl alcohol respectively. Observed percent of analytical recovery for bile acid estimation was <98% for activated polymeric membrane bound bile acid enzymes. Improved % of retention activities were observed for activated polymeric membranes bound bile acid enzymes in our present method even after the regular use of more than 4 months (16 weeks) when preserved at 4 dgree C. Hence, our present method of immobilization of bile acid enzymes onto activated polymeric membranes were found to be more cost effective as compared to other reported enzyme based methods.