Tannase immobilisation by amino-functionalised magnetic Fe3O4-chitosan nanoparticles and its application in tea infusion

The tannase (from Aspergillus niger ) was immobilised by glutaraldehyde conjugation to amino-functionalised chitosan-coated magnetic nanoparticles (Fe 3 O 4 -CS nanoparticles). Fourier-transform infrared spectroscopy and thermo-gravimetric analysis showed that chitosan was coated on the surface of magnetic nanoparticles. Transmission electron microscopy indicated that the synthesised nanoparticles (Fe 3 O 4 -CS) were almost spherical or ellipsoidal with an average diameter of 5.97 ± 1.25 nm. The stability and functionality of free and immobilised tannase were compared. Both forms of tannase exhibited the same optimal temperature of 30 °C, whereas the optimal pH value of immobilised tannase (pH 4.5) was lower than that of the free tannase (pH 5.5). The pH and thermal stabilities of immobilised tannase were significantly better than those of free tannase. Immobilised tannase retained over 50% of its initial activity after repeated utilisation for eight cycles. Furthermore, the immobilised tannase effectively improve the clarity and colour of black and green tea infusions. These results showed that amino-functionalised Fe 3 O 4 -CS nanoparticles are an efficient carrier for immobilising tannase, and immobilised tannase can be used in the clarification of tea infusion.