| The use of many enzymes in industrial applications requires high thermostability. Researchers used diversified bioinformatic tools to simulate natural evolution in enzyme design. They are able to choose between three different, apparently equally successful, strategies: rational design, directed evolution and semi-rational methods.In this paper, we studied the thermostability of CALB and got three different mutated enzymes, CALB5, CALB7 and M168, which are more thermostable than CALB. First, we used homology modeling to predict the 3D-structure of enzymes. After that, we performed molecular docking to simulate the connection between receptor and ligand. Finally, we tried Molecular Dynamics Simulation to simulate the moving trajectory of enzymes in water solution and calculated the parameters related to thermostability.We analyzed the mechanism of improved thermostability in the aspects of 3D-structure, Total Energy and RMSD by using computer-assisted methods.We testedβ-glucuronidase(GUS), Aspergillus niger phytase (PhyA), E.coli AppA2 phytase, Bacillus subtilis cold shock proteins(Csp) and human adenosine A2a receptor(Asp) containing 32 mutants using Molecular Dynamics Simulations and analyzed the relationship between thermostability and Total Energy & RMSD. Our prediction achieved 88% accuracy and a correlation coefficient is more than 0.8.On the other hand, we calculated the B-factor of all amino acids in CALB using B-FITTER and changed several sites with higher B-factor. Finally, we designed a mutant with higher thermostability called CALBM. We discovered CALBM is more thermostable experimentally compared with CALB.Bioinformatics and the development of computer science have opened up new prospects for biological. The use of large-scaled high-performance computers and related software made it possible to perform biological experiments by computer-assisted methods which greatly improve the experimental efficiency and reduce cost effectively in the future.
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