Fabrication of gold nanoparticle assembled polyurethane microsphere template in trypsin immobilization

The separation, reusability and high catalytic activity of bioconjugate remain challenging task in proteins bound gold nanoparticles. A facile synthetic route for the fabrication of gold nanoparticle assembled polyurethane microsphere template and immobilization of trypsin on gold/polyurethane surface to form trypsin-nanogold-polyurethane bioconjugate was developed. The bioconjugate was characterized by X-ray diffraction, Field emission scanning electron microscopy, Transmission electron microscopy, UV-visible, Fourier transform infrared, Fluorescence and Circular dichroism spectroscopy. The catalytic studies confirmed retention of approximately 40% of its original activity even after eight consecutive reaction cycles. The bioconjugate is also very effective for its separation from the reaction medium and exhibited significant enhanced stability over a wide range of pH and temperature compared to free trypsin. These findings clearly demonstrate that trypsin immobilized gold nanoparticle assembled polyurethane microsphere acts as an excellent recyclable biocatalyst with enzyme-specific biocompatibility.