Semirational Engineering of <i>Clostridium thermosuccinogenes</i> <scp>l</scp> -Rhamnose Isomerase by Targeting Flexible Regions Adjacent to the Active Pocket for <scp>d</scp> -Allose Biosynthesis

d-allulose. Molecular dynamics (MD) simulations revealed that modulation of loop flexibility enhanced the enzyme turnover rate. By fine-tuning loop flexibility while maintaining the structural integrity of the enzyme, this engineered mutant reduces the risk of enzyme inactivation during prolonged high-temperature catalysis. Collectively, these results highlight its potential for industrial d-allose production and provide valuable insights for the modification of L-RIs and related enzymes.