Engineering of β-Agarase with Enhanced Thermostability via Multitool Consensus Prediction and Structure-Guided Screening

) at 50 °C than the wild-type. Molecular dynamics simulations showed that mutations strengthened hydrophobic interactions, salt bridges, and hydrogen bond networks, while optimizing surface charge. The M3 variant performed well in high-temperature agarose hydrolysis, mainly producing neoagarotetraose (NA4) and neoagarohexaose (NA6), showing great industrial potential. This framework improves the efficiency of enzyme thermostability engineering and provides a general approach for industrial enzymes modification.