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In-Silico Side-Directed Mutagenesis of Oxidoreductase from Anoxybacillus sp. SK3-4

Received: 27 June 2018     Accepted: 11 July 2018     Published: 9 August 2018
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Abstract

Precisely, mutagenesis can introduce mutations into the target gene by using mismatch primers which are partially complementary to the template strand of DNA using polymerase chain reaction (PCR). Oxidoreductase enzymes are generally proteins that involved in oxidation-reduction reactions in biological systems. For this study, primary sequence analyses of oxidoreductase protein from Anoxybacillus sp. SK3-4 was conducted with the aim of generating functional information and theoretically improve catalytic stability of the protein by in-silico mutagenesis. The primary sequence of a novel protein with 386 amino acid residues was analyzed using Expasy-tool for translation of the amino acid sequence into a nucleotide gene sequence. Important catalytic binding sites of the protein were predicted using 3DLigandSite program, Pheres2 and Protein Bioedit servers for generating functional information of the protein. Site-directed mutagenesis (SDM) was used against the novel protein (oxidoreductase), in which two site mutations were created based on rational design. Amino acids; leucine (L) and histidine (H), involved in substrate and metal binding sites in the protein were substituted for isoleucine (I) and arginine (R) i.e. L138I and H280R, to check for significant change in the functional stability of the protein, thereby increasing the efficiency of the enzyme to help speed up the rate of chemical reactions.

Published in International Journal of Microbiology and Biotechnology (Volume 3, Issue 2)
DOI 10.11648/j.ijmb.20180302.14
Page(s) 51-56
Creative Commons

This is an Open Access article, distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution and reproduction in any medium or format, provided the original work is properly cited.

Copyright

Copyright © The Author(s), 2018. Published by Science Publishing Group

Keywords

Anoxybacillus sp. SK3-4, Oxidoreductase, Catalytic Stability, Site-Directed Mutagenesis

References
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Cite This Article
  • APA Style

    Musa Ahmed Abubakar, Umar Shittu, Musa Hassan Muhammad, Saif Al-Sheyab Ahmad. (2018). In-Silico Side-Directed Mutagenesis of Oxidoreductase from Anoxybacillus sp. SK3-4. International Journal of Microbiology and Biotechnology, 3(2), 51-56. https://doi.org/10.11648/j.ijmb.20180302.14

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    ACS Style

    Musa Ahmed Abubakar; Umar Shittu; Musa Hassan Muhammad; Saif Al-Sheyab Ahmad. In-Silico Side-Directed Mutagenesis of Oxidoreductase from Anoxybacillus sp. SK3-4. Int. J. Microbiol. Biotechnol. 2018, 3(2), 51-56. doi: 10.11648/j.ijmb.20180302.14

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    AMA Style

    Musa Ahmed Abubakar, Umar Shittu, Musa Hassan Muhammad, Saif Al-Sheyab Ahmad. In-Silico Side-Directed Mutagenesis of Oxidoreductase from Anoxybacillus sp. SK3-4. Int J Microbiol Biotechnol. 2018;3(2):51-56. doi: 10.11648/j.ijmb.20180302.14

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  • @article{10.11648/j.ijmb.20180302.14,
      author = {Musa Ahmed Abubakar and Umar Shittu and Musa Hassan Muhammad and Saif Al-Sheyab Ahmad},
      title = {In-Silico Side-Directed Mutagenesis of Oxidoreductase from Anoxybacillus sp. SK3-4},
      journal = {International Journal of Microbiology and Biotechnology},
      volume = {3},
      number = {2},
      pages = {51-56},
      doi = {10.11648/j.ijmb.20180302.14},
      url = {https://doi.org/10.11648/j.ijmb.20180302.14},
      eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.ijmb.20180302.14},
      abstract = {Precisely, mutagenesis can introduce mutations into the target gene by using mismatch primers which are partially complementary to the template strand of DNA using polymerase chain reaction (PCR). Oxidoreductase enzymes are generally proteins that involved in oxidation-reduction reactions in biological systems. For this study, primary sequence analyses of oxidoreductase protein from Anoxybacillus sp. SK3-4 was conducted with the aim of generating functional information and theoretically improve catalytic stability of the protein by in-silico mutagenesis. The primary sequence of a novel protein with 386 amino acid residues was analyzed using Expasy-tool for translation of the amino acid sequence into a nucleotide gene sequence. Important catalytic binding sites of the protein were predicted using 3DLigandSite program, Pheres2 and Protein Bioedit servers for generating functional information of the protein. Site-directed mutagenesis (SDM) was used against the novel protein (oxidoreductase), in which two site mutations were created based on rational design. Amino acids; leucine (L) and histidine (H), involved in substrate and metal binding sites in the protein were substituted for isoleucine (I) and arginine (R) i.e. L138I and H280R, to check for significant change in the functional stability of the protein, thereby increasing the efficiency of the enzyme to help speed up the rate of chemical reactions.},
     year = {2018}
    }
    

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  • TY  - JOUR
    T1  - In-Silico Side-Directed Mutagenesis of Oxidoreductase from Anoxybacillus sp. SK3-4
    AU  - Musa Ahmed Abubakar
    AU  - Umar Shittu
    AU  - Musa Hassan Muhammad
    AU  - Saif Al-Sheyab Ahmad
    Y1  - 2018/08/09
    PY  - 2018
    N1  - https://doi.org/10.11648/j.ijmb.20180302.14
    DO  - 10.11648/j.ijmb.20180302.14
    T2  - International Journal of Microbiology and Biotechnology
    JF  - International Journal of Microbiology and Biotechnology
    JO  - International Journal of Microbiology and Biotechnology
    SP  - 51
    EP  - 56
    PB  - Science Publishing Group
    SN  - 2578-9686
    UR  - https://doi.org/10.11648/j.ijmb.20180302.14
    AB  - Precisely, mutagenesis can introduce mutations into the target gene by using mismatch primers which are partially complementary to the template strand of DNA using polymerase chain reaction (PCR). Oxidoreductase enzymes are generally proteins that involved in oxidation-reduction reactions in biological systems. For this study, primary sequence analyses of oxidoreductase protein from Anoxybacillus sp. SK3-4 was conducted with the aim of generating functional information and theoretically improve catalytic stability of the protein by in-silico mutagenesis. The primary sequence of a novel protein with 386 amino acid residues was analyzed using Expasy-tool for translation of the amino acid sequence into a nucleotide gene sequence. Important catalytic binding sites of the protein were predicted using 3DLigandSite program, Pheres2 and Protein Bioedit servers for generating functional information of the protein. Site-directed mutagenesis (SDM) was used against the novel protein (oxidoreductase), in which two site mutations were created based on rational design. Amino acids; leucine (L) and histidine (H), involved in substrate and metal binding sites in the protein were substituted for isoleucine (I) and arginine (R) i.e. L138I and H280R, to check for significant change in the functional stability of the protein, thereby increasing the efficiency of the enzyme to help speed up the rate of chemical reactions.
    VL  - 3
    IS  - 2
    ER  - 

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Author Information
  • Department of Science Laboratory Technology, Kano State Polytechnic, Kano, Nigeria

  • Department of Biology, Isa Kaita College of Education, Dutsin-ma, Katsina, Nigeria

  • Department of Microbiology & Biotechnology, Federal University Dutse, Gigawa, Nigeria

  • Department of Applied Biological Sciences & Arts, Jordan University of Science and Technology, Irbid, Jordan

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