Examination of the effect of pH and temperature on the activity of nanosystem containing chondroitinase ABCI based on hydroxyapatite

Chondroitinase ABCI is a bacterial lyase that degrades glycosaminoglycans and promotes axonal growth and functional improvement. However the stability and maintenance of this enzyme is very limited. One of the strategies to overcome this limitation is to immobilize the enzyme. In this research, chondroitinase ABCI (cABCI) from Proteus Vulgaris was immobilized on hydroxyapatite nanoparticles. Hydroxyapatite is a non-toxic ceramic biomaterial that has a high surface area, which is beneficial for loading a large amount of enzyme. Therefore, to increase the stability of chondroitinase ABCI, immobilization on hydroxyapatite nanoparticles for 4 hours through physical adsorption in  phosphate buffer pH 5, 6.8, and 8 at 4◦C was carried out. Enzyme immobilization on hydroxyapatite nanoparticles was then confirmed by field emission gun-scanning electron microscopy and UV-spectroscopy, before and after immobilization. Then, in order to obtain the optimal pH and temperature, the activity of the nanosystem was investigated at three pH and temperatures (4°C, 25°C, and 37°C). Results revealed higher activity at pH 5 and temperature 4 ◦C than the other pH and temperatures for the nanosystem. Based on the obtained results, which show the stability of the nanosystem at all three temperatures compared to the free enzyme, this nanosystem could be a potential candidate for clinical applications in future.