Mechanism of Hyaluronic Acid Hydrolysis Catalyzed by Snake Venom Hyaluronidase

. Our simulations also support an induced-fit mechanism for the svHyal/hyaluronic acid complex, with substrate distortion (chair → boat/skew-boat) favoring a conformation that closely mimics the transition state. This distortion, along with a prealignment of Glu112, lowers the activation free energy, enhancing the susceptibility of the glycosidic bond to nucleophilic attack. The results are likely transferable to all svHyal given their high degree of interspecific similarity (>90% sequence identity). This study highlights the importance of understanding mechanistics, including detailed stereoelectronic conformations and subsite-specific interactions, for the design of novel and effective inhibitors with broad clinical and biotechnological applications.