Preparation and properties of thermoreversible, phase-separating enzyme-oligo(N-isopropylacrylamide) conjugates.

A thermoreversible N-isopropylacrylamide (NIPAAm) oligomer with an N-hydroxysuccinimide (NHS) ester functional end group has been prepared for coupling to an enzyme, beta-D-glucosidase, to form a thermoreversible, phase-separating polymer-enzyme conjugate. This conjugate can be used for separation, recovery, and recycle of an enzyme simply by applying small temperature changes to the reaction medium. In contrast to the random polymer-enzyme conjugates previously reported by us and others in the literature, in this study the conjugate is formed by a single, end attachment of each oligomer chain to the enzyme. Preliminary studies show that the conjugated enzyme exhibits very high retention of activity, even higher than native enzyme, and shows improved thermal stability compared to native enzyme.