The role of proline substitutions within flexible regions on thermostability of luciferase.Improving the stability of firefly luciferase has been a critical issue for its wider industrial applications.  Studies about hyperthermophile proteins show that flexibility could be an effective indicator to find out weak spots to engineering thermostability of proteins.  Proline is the most rigid residue and can be introduced to rigidify flexible regions to enhance thermostability of proteins.  In the results, H489P mutant shows improved thermostability while maintaining its catalytic efficiency compared to that of wild type luciferase.  D476P mutant shows decreased thermosatbility and the reason for this is elucidated at the molecular level.  Thermostability analysis shows that S307P mutation has decreased kinetic stability and enhanced thermodynamic stability.
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