Hyaluronidase enzyme conjugated polyamidoamine dendrimer: An efficient and stable nanobiocatalyst for enzymatic degradation of hyaluronic acid

In this work hyper-branched poly (amide amine) (PAMAM) nanoparticles were conjugated with hyaluronidase (Hyal) to produce a robust nano-biocatalyst for hyaluronic acid (HA) degradation. The success enzyme attachment process was confirmed by Fourier transform infrared (FTIR), transmission electron microscopy (TEM), dynamic light scattering (DLS), and UV-Vis. The influence of pH, temperature, and inhibitor on the enzymatic activity of hyaluronidase was also investigated. The optimum pH, temperature and storage time of Hyal-PAMAM nanocomplex were higher than free enzyme. Also, ascorbic acid showed more inhibitory effect on free enzyme, the IC50 values were determined to be around 55±0.7 and 70±0.3 mM for free Hyal and Hyal-PAMAM nanocomplex, respectively. Based on the greater υmax and lower Km, the Hyal-PAMAM showed a better catalytic efficiency for HA degradation. Moreover, the in silico screening of PAMAM/Hyal interactions further confirmed the experimental results. The novel strategy for combining Hyal and PAMAM dendrimer can hold great promise for applications in biomedical, sensing, and industrial catalysis.