Volume 22, Issue 5 (11-2019)                   J Arak Uni Med Sci 2019, 22(5): 68-77 | Back to browse issues page

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Fallahzadeh R, Esfahani K, Akhavan Sepahi A, Kamali N, Bambai B. Increasing the catalytic power of the flavin reductase DszD enzyme using site-directed mutagenesis method in Rhodococcus erythropolis. J Arak Uni Med Sci 2019; 22 (5) :68-77
URL: http://jams.arakmu.ac.ir/article-1-6094-en.html
1- Department of Microbiology, Tehran North Branch of Islamic Azad University, Tehran, Iran.
2- Department of Medical Genetics, Institute of Medical Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran.
3- Department of Medical Genetics, Institute of Medical Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran. , bambai@nigeb.ac.ir
Abstract:   (2923 Views)
Background and Aim The flavin reductase DszD enzyme is a key enzyme for providing required reduction potential in the bacterial desulfurization process. Considering the low speed of desulfurization process because of low catalytic power of this enzyme, it is necessary to increase the catalytic power of flavin reductase for industrial use of this enzyme as biocatalyst.
Methods & Materials The three-dimensional structure of the flavin reductase DszD enzyme was predicted by a CPHmodel server and its amino acid sequence was searched in the protein data bank to identify the homologue molecules. Based on the alignment of the amino acid sequence and the model molecules, the key residues at the flavin mononucleotide substrate were identified. The key residue of asparagine at position 77 was replaced with phenylalanine using the site-directed mutagenesis method. 
Ethical Considerations This study with research ethics code IR.NIGEB.EC.1398.6.24 A has been approved by research ethics committee at National Institute of Genetic Engineering and Biotechnology, Tehran, Iran.
Results The cloning and expression of each of the wild-type and mutant genes were performed separately. The catalytic power of the produced wild-type and mutant enzymes were compared. The catalytic activity measurements showed that the mutant enzyme had a 2.5 fold increase in catalytic power.
Conclusion Replacing phenylalanine with asparagine at position 77 of flavin reductase DszD enzyme leads to an increase in enzyme catalytic power to increase the speed of bacterial desulfurization process.
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Type of Study: Original Atricle | Subject: Basic Sciences
Received: 2019/06/13 | Accepted: 2019/10/14

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