4.7 Article

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

Journal

JOURNAL OF MOLECULAR LIQUIDS
Volume 349, Issue -, Pages -

Publisher

ELSEVIER
DOI: 10.1016/j.molliq.2021.118111

Keywords

Hyaluronidase; PAMAM dendrimer; Enzyme immobilization; Hyaluronic acid; Enzymatic degradation; Inhibitory activity

Funding

  1. Research Council of University of Isfahan
  2. Biotechnology Development Council of the Islamic Republic of Iran [970704]
  3. Australian Research Council Centre Discovery Early Career Researcher Award (DECRA) [DE180100688]

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In this study, hyper-branched poly (amide amine) (PAMAM) nanoparticles were conjugated with hyaluronidase (Hyal) to create a stable nano-biocatalyst for hyaluronic acid (HA) degradation. The results showed that the Hyal-PAMAM nanocomplex had higher enzymatic activity at optimum pH, temperature, and storage time compared to free enzyme. In addition, computational screening further confirmed the experimental results. This strategy of combining Hyal and PAMAM holds great promise for applications in biomedical, sensing, and industrial catalysis.
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 upsilon(max) and lower K-m, 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. (C) 2021 Elsevier B.V. All rights reserved.

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