Enzyme modification by polymers with solubilities that change in response to photoirradiation in organic media.

Abstract

We have synthesized a hybrid subtilisin the solubility of which can be regulated by photoirradiation through coupling with a photoresponsive copolymer that carries spiropyran groups in its side chains. The copolymer was synthesized by polymerization of methacrylate, methacrylic acid, and spiropyran-carrying methacrylate. It was then covalently bonded to the amino groups of subtilisin Carlsberg via its carboxyl groups using a carbodiimide coupling agent. The hybrid subtilisin was perfectly soluble in toluene and efficiently catalyzed transesterification. After ultraviolet irradiation, the hybrid subtilisin precipitated and was easily and quantitatively recovered by centrifugation. Recovered hybrid subtilisin, resolubilized by visible light irradiation, retained its original transesterification activity even after several cycles of precipitation and solubilization.