Encapsulation of Enzymes in Biodegradable Tubular Structures

Abstract

Enzymes were directly encapsulated in microtubes fabricated by coelectrospinning. Coelectrospinning makes it possible to separate between an organic phase, which may be harmful to enzyme activity, and an aqueous one. In this report, enzymes were incorporated in an aqueous core solution of poly(ethylene oxide) (PEO) while the shell solution was made of polycaprolactone (PCL) dissolved in a mixture of chloroform and dimethylformamide (DMF). Indeed, this separation was found to preserve the enzymatic activities of two enzymes during and after electrospinning, as ascertained when the fiber mats were placed in an aqueous assay environment. The shell morphology was altered by blending PCL with poly(ethylene glycol) (PEG). This blending induced pores in the shell which in turn pronouncedly affected the movement of molecules into or out of the fibers. An enzyme of about 80 kDa, alkaline phosphatase (AP), diffused through the pores into the surrounding medium. The fibers can thus act as release devices. However, a larger enzyme, β-galactosidase (465 kDa), remained in the fibers without any leaching. Enzyme assays showed that the substrates of both enzymes diffused efficiently into the fibers and were cleaved by the encapsulated enzyme, and the reaction product then diffused outside. With encapsulated β-galactosidase, the fiber acted as an efficient enzymatic microreactor and the relative enzymatic activity reached a level of about 50%.