biostudies-literatureXi KWarshel Institute for Computational BiologyNational Science Foundation of ChinaShenzhen Fundamental Research FundShenzhen-Hong Kong Cooperation Zone for Technology and Innovationthe National Science Foundation of China14628https://www.ebi.ac.uk/biostudies/studies/S-EPMC9736071biostudies-literatureUnknown23(23)Bacteriophage T4 lysozyme (T4L) is a glycosidase that is widely applied as a natural antimicrobial agent in the food industry. Due to its wide applications and small size, T4L has been regarded as a model system for understanding protein dynamics and for large-scale protein engineering. Through structural insights from the single conformation of T4L, a series of mutations (L99A,G113A,R119P) have been introduced, which have successfully raised the fractional population of its only hydrolysis-competent excited state to 96%. However, the actual impact of these substitutions on its dynamics remains unclear, largely due to the lack of highly efficient sampling algorithms. Here, using our recently developed travelling-salesman-based automated path searching (TAPS), we located the minimum-free-enInternational journal of molecular sciencesAutomated Path Searching Reveals the Mechanism of Hydrolysis Enhancement by T4 Lysozyme Mutants.PMC973607131971179JCYJ20200109150003938HZQB-KCZYB-2020056RCYX20200714114645019Xi KZhu LfalseAutomated Path Searching Reveals the Mechanism of Hydrolysis Enhancement by T4 Lysozyme Mutants.Bacteriophage T4 lysozyme (T4L) is a glycosidase that is widely applied as a natural antimicrobial agent in the food industry. Due to its wide applications and small size, T4L has been regarded as a model system for understanding protein dynamics and for large-scale protein engineering. Through structural insights from the single conformation of T4L, a series of mutations (L99A,G113A,R119P) have been introduced, which have successfully raised the fractional population of its only hydrolysis-competent excited state to 96%. However, the actual impact of these substitutions on its dynamics remains unclear, largely due to the lack of highly efficient sampling algorithms. Here, using our recently developed travelling-salesman-based automated path searching (TAPS), we located the minimum-free-en2022-01-01T00:00:00Z2022 Nov2026-04-08T14:25:58.118Z2025-04-04T23:37:09.431ZS-EPMC97360713649895410.3390/ijms232314628