Identification of Lampyris turkestanicus luciferase Stabilizing Mutations Using Bioinformatics Software and Web Servers and Molecular Dynamics Simulation

Subject Areas : bioinformatics

Alireza Khondabi ¹ (Department of Genetics and Biotechnology, School of Biological Science, Varamin-Pishva Branch, Islamic Azad University, Varamin,Iran)
hassan sahebjamee ² (Department of Biophysics, School of Biological Science, Varamin-Pishva Branch, Islamic Azad University, Varamin,Iran)
Fahimeh Baghbani-Arani ³ (Department of Genetics and Biotechnology, School of Biological Science, Varamin-Pishva Branch, Islamic Azad University, Varamin,Iran)

Received: 2021-08-06 Accepted : 2021-09-09 Published : 2020-11-21

Keywords: Stability, Mutation, molecular dynamics, Luciferase,

Abstract :

The conversion of chemical energy to light by a living organism, bioluminescence, is an alluring process that catalyzes by enzymes generically called luciferases. This enzyme has wide applications as a reporter gene in in vivo imaging. In spite of wide range of luciferase applications, some inherent properties limit further application and development of this technology, including the low stability of the enzyme, a low turnover number, and a high Km for the substrate ATP.Due to the low stability of luciferase enzyme, the present study was performed to evaluate the effect of single amino acid mutations on the stability of luciferase enzyme of Iranian species Lamphyris Turkestanius using bioinformatics web servers and molecular dynamics simulation.For this purpose, to determine the stability, the mutations were tested using I-Mutant-2, Pop-music, Cupsat, istable, MUpro and Protparam and foldx web servers under Yasara software. Web servers and foldx predicted that two mutations, Q35L and H9M, would stabilize the structure. The web server Polyphen-2 predicted that these two mutations would not have a damaging effect on enzyme function. To further confirm these two mutations, molecular dynamics simulations were performed. The results of RMSD and RMSF and the radius of gyrus and hydrophobicity also showed that these mutations may cause increase the stability of this enzyme.