Abstract

A hydbrid type of metalloenzyme membrane has been applied to flow-amperometric biosensing of multi-ions based on the apoenzyme reactivation method. The hybrid-enzyme membrane prepared by co-immobilizing metal-depending enzymes, i.e., alkaline phosphatase and ascorbate oxidase, onto a partially aminated polyacrylonitrile membrane is used as the recognition element for zinc(II), cobalt(II) and copper(II) ions. This biosensing system is assembled with the enzyme membrane tightly attached onto a flow-through oxygen electrode for monitoring the enzymic activity. Biosensing of zinc(II), cobalt(II) and copper(II) ions at submillimolar levels has been amperometrically realized through the activation of each of the immobilized metal-free enzymes (apoenzymes).

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