IN SITU INVESTIGATION OF TYROSINASE IMMOBILIZATION ON POLY-(ACRYLONITRILE- CO-ACRYLAMIDE) BY QUARTZ CRYSTAL MICROBALANCE
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2021-01-01
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Abstract
In this paper, we report the results of the covalent immobilization of tyrosinase onto polymer based on poly-(acrylonitrile-co-acrylamide) poly-(AN-co-AM) using two methods - in a microfluidic cell with quartz crystal microbalance (QCM) detection and in self-constructed laboratory immobilizer. Poly-(AN-co-AM) is a well-known polymer for enzyme immobilization which is low cost, easy for manipulation and with good chemical stability. This polymer is also biocompatible, insoluble in water, contains suitable functional groups for covalent immobilization, and can easily be deposited as thin layer. The results from Fourier-transform infrared spectroscopy show two sharp absorption peaks at 2243 cm-1 and 1690 cm-1 are attributed to stretching of -CN and -CONH2 groups, respectively, and are related to the covalent binding between the enzyme and the polymer. Spin-coating method was used for thin film deposition. The prepared polymer layer was less than 100 nm thickness. A QCM was used to monitor the amount of protein attached to the sensor surface during the laminar liquid flow in the microfluidic cell. Scanning Electron Microscopy and Atomic Force Microscopy show the stages of covering of quartz resonator surface by enzyme.