Evolved enzymes in the metabolism of biological poly-acids: Applications in otolaryngological biocatalysis

This study explores evolved Hyaluronidase, Lipase, and Elastase's identification, characterization, and therapeutic potential to enhance tissue regeneration and drug delivery systems in otolaryngology. Hyaluronidase variant H5 exhibited a turnover number (k_cat) of 1500 min^-1, a 200 % increase over wild-type (500 min^-1), demonstrating superior hyaluronic acid degradation. Similarly, lipase variant L2 reached 1200 min^-1 (400 min^-1 wild-type), and elastase variant E3 showed a turnover of 2200 min^-1 (1000 min^-1 wild-type). Kinetic analyses revealed improved Km and Vmax values across variants, with Hyaluronidase Variant H5 achieving Km = 1.5 μM and Vmax = 3000 μM/min. Molecular Dynamics (MD) simulations indicated structural stability (average RMSD ~1.5 Å for H5) and strong hydrogen bonding (180 bonds), enhancing catalytic efficiency. In vitro assays demonstrated a 40 % enhancement in tissue regeneration and increased epithelial cell proliferation (100 % for Hyaluronidase Variant H5 vs. 60 % wild-type). In vivo studies in rabbits revealed a 30 % reduction in recovery time post-sinus surgery and a 50 % reduction in scar tissue formation. These findings underscore the potential of evolved enzymes in advancing drug delivery (DD) and tissue repair (TR), with implications for broader applications in wound healing and inflammatory diseases.