International Journal of Microbiology and Biotechnology
Volume 3, Issue 2, June 2018, Pages: 51-56
Received: Jun. 27, 2018;
Accepted: Jul. 11, 2018;
Published: Aug. 9, 2018
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Musa Ahmed Abubakar, Department of Science Laboratory Technology, Kano State Polytechnic, Kano, Nigeria
Umar Shittu, Department of Biology, Isa Kaita College of Education, Dutsin-ma, Katsina, Nigeria
Musa Hassan Muhammad, Department of Microbiology & Biotechnology, Federal University Dutse, Gigawa, Nigeria
Saif Al-Sheyab Ahmad, Department of Applied Biological Sciences & Arts, Jordan University of Science and Technology, Irbid, Jordan
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.
Musa Ahmed Abubakar,
Musa Hassan Muhammad,
Saif Al-Sheyab Ahmad,
In-Silico Side-Directed Mutagenesis of Oxidoreductase from Anoxybacillus sp. SK3-4, International Journal of Microbiology and Biotechnology.
Vol. 3, No. 2,
2018, pp. 51-56.
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