Molecular Engineering of the Geobacillus stearothermophilus α-Amylase and Cel5E from Chlostridium thermocellim; In Silico Approach
Article 3, Volume 16, Issue 3, Summer 2018, Page 185-192  XMLPDF (3367 K)
Document Type: Research Paper
DOI: DOI:10.15171/IJB.1284
Authors
Ibrahim Torktaz1, 2; Jafar Hemmat  2; Ali Asghar Karkhane1; Garshasb Rigiorcid 3; Amin Rostami4; Jafar Khezri1; Reza Behroozi1
1National Institute of Genetic Engineering and Biotechnology, Tehran, Iran
2Biotechnology Department, Iranian Research Organization for Science and Technology (IROST) Tehran-Iran.
3Department of Genetics, Faculty of Basic Science, University of Shahrekord, Shahrekord, Iran
4Department of Basic Sciences, Faculty of Paramedical Sciences, Shahid Beheshti University of Medical Sciences, Tehran, Iran
Abstract
Background: Considering natural thermal stability, Geobacillus stearothermophilus amylase and Cel5E from Clostridium thermocellum are good candidates for industrial applications. To be compatible with the industrial applications, this enzyme should be stable in the high temperatures, so any improvement in their thermal stability is valuable.
Objectives: Using in silico approach and identifying point mutations in the structure amylase of G. stearothermophilus and Cel5E from C. termocellum we tried to increase thermal stability of the enzymes along with their catalytic activity to reach a new industrial amylase with higher thermostability and an improved function.
Materials and Methods: In this study we predicted the 3D structure of the enzymes, then simulated the molecular docking study using MolDock, PLANTS, and Lamarkian genetic algorithm as scoring functions for the docking and in silico engineering of the protein aiming to increase the thermal stability and catalytic activity.
Results: A series of thermal stability increasing point mutations were exerted around the active site of the enzyme, then by docking procedure, the binding affinity was measured and finally a list of mutations which theoretically improved the increased thermal stability as well as catalytic activity were proposed.
Conclusions: Based on the in silico results obtained the modified enzymes seems to be suitable candidates for considering in both laboratory and industrial scales.
Keywords
Amylase; Catalytic activity; Cel5E; Geobacillus stearothermophilus; Industry; Thermal stability
Main Subjects
Bio-informatics
Supplementary Files