This Brief highlights different approaches used to create stable cellulase and its use in different fields. Cellulase is an industrial enzyme with a broad range of significant applications in biofuel production and cellulosic waste management. Cellulase 7a from Trichoderma reesei is the most efficient enzyme in the bio hydrolysis of cellulose. In order to improve its thermal stability, it can be engineered using a variety of approaches, such as hydrophobic interactions, aromatic interactions, hydrogen bonds, ion pairs and disulfide bridge creation.
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1 Introduction of Cellulase and Its Application |
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1 | (6) |
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1.1 Biofuel as a Suitable Alternative for Fossil Fuel |
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1.2 Enzyme Stability Is One of the Limits in Biofuel Production |
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1.3 A Procedure to in Silico Engineering of Cellulase |
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1.4 Cellulase 7a Selected as a Target |
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1.5 Advantages of Cellulose in Silico Engineering |
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2 Literature Review of Cellulase and Approaches to Increase Its Stability |
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2.1 Cellulose and Cellulases Structure |
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2.2 Applications of Cellulase |
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2.2.1 Application of Cellulase in Biofuel Production |
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2.3 Trichoderma reesei Known as the Crowned King of Cellulolytic Fungi |
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2.4 Catalytic Mechanisms of Glycoside Hydrolases |
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2.5 An Introduction to Cellulase Family 7 |
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2.6 Cellulose Biodegradation Procedures |
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2.7 Protein Engineering as a Solution |
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2.7.1 Rational Design Approach |
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2.8 Advantages of Enzyme Thermo Stabilization |
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2.9 Some Thermo Stabilize Mutations |
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2.9.1 Introducing Disulfide Bonds as a Strategy to Increase Stability |
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2.10 Molecular Dynamic Simulation as a Strong Approach to Evaluating Structure Stability |
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3 Methodology of Mutant Creation and Molecular Dynamic Simulation |
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3.1 Obtaining Protein Structure File |
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3.2.1 Removing Water Molecule and Ligands |
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3.3 Generation of Mutated pdb Files |
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3.4 Molecular Dynamic Simulation Steps |
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3.4.1 pdb2gmx (Topology File Generation) Tool |
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3.4.2 Unit Cell Defining the by the Tools, editconf and Adding Solvent |
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3.4.4 Energy Minimization as a Critical Step |
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3.4.5 Equilibration of Temperature and Pressure |
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3.5 Tools for Analysis of Molecular Dynamics Trajectory |
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4 Result and Discussion of Molecular Dynamic Simulation of Created Mutants |
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4.1 Nominated Residues for Mutation |
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4.2 Native Protein Structure and Stability Analysis |
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4.3 Mutated Protein Number 1 (E385C and A392C) Structure and Stability Analysis |
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4.4 Mutated Protein Number 2 (Y321C and A333C) Structure and Stability Analysis |
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4.5 Mutated Protein Number 3 (T383C and T399C) Structure and Stability Analysis |
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4.6 Mutated Protein Number 4 (A187C) Structure and Stability Analysis |
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4.7 Mutated Protein Number 5 (D257C and L346C) Structure and Stability Analysis |
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5 Conclusion of Simulation Analysis of Mutants |
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5.1 Conclusion of MD Simulation of Created Mutants |
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5.2 Future Work of the Disulfide Bond Engineering to Cellulase |
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Dr. I. S. Amiri, received his B. Sc (Hons, Applied Physics) from Public University of Orumiyeh, Iran in 2001 and a gold medalist M. Sc. from Universiti Teknologi Malaysia (UTM), in 2009. He was awarded a PhD degree in nano photonics in 2013. He has published more than 200 journals/conferences and books in Optical Soliton Communications, Laser Physics, Photonics, Optics, Nanophotonics, Nonlinear fiber optics, Quantum Cryptography, Optical Tweezers, Nanotechnology, Biomedical Physics and Biotechnology Engineering. Now he is a visiting research fellow in University of Malaya, 50603 Kuala Lumpur, Malaysia.
Mr. B. Barati received his B. Sc (cellular and molecular Genetics) from Azad University of Tonekabon, Iran in 2011 and he is recently graduated from M. Sc. in Biotechnology from Universiti Teknologi Malaysia (UTM), in 2013. His research interests are in the field of protein engineering, enzyme production, Genetic engineering, drug design, molecular dynamic simulation and bioinformatics.
Lisainfo e-raamatute kohta