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Molecular characterization of a recombinant managanese superoxide dismutase from Lactococcus lactis M4


Tan, Boon Hooi and Leow, Adam Thean Chor and Foo, Hooi Ling and Abdul Rahim, Raha (2014) Molecular characterization of a recombinant managanese superoxide dismutase from Lactococcus lactis M4. BioMed Research International, 2014. art. no. 469298. pp. 1-9. ISSN 2314-6133; ESSN: 2314-6141


A superoxide dismutase (SOD) gene of Lactococcus lactis M4 was cloned and expressed in a prokaryotic system. Sequence analysis revealed an open reading frame of 621 bp which codes for 206 amino acid residues. Expression of sodA under T7 promoter exhibited a specific activity of 4967 U/mg when induced with 1 mM of isopropyl-β-D-thiogalactopyranoside. The recombinant SOD was purified to homogeneity by immobilised metal affinity chromatography and Superose 12 gel filtration chromatography. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and western blot analyses of the recombinant SOD detected a molecular mass of approximately 27 kDa. However, the SOD was in dimer form as revealed by gel filtration chromatography. The purified recombinant enzyme had a pI of 4.5 and exhibited maximal activity at 25°C and pH 7.2. It was stable up to 45°C. The insensitivity of this lactococcal SOD to cyanide and hydrogen peroxide established that it was a MnSOD. Although it has 98% homology to SOD of L. lactis IL1403, this is the first elucidated structure of lactococcal SOD revealing active sites containing the catalytic manganese coordinated by four ligands (H-27, H-82, D-168, and H-172).

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Additional Metadata

Item Type: Article
Divisions: Faculty of Biotechnology and Biomolecular Sciences
Institute of Bioscience
DOI Number: https://doi.org/10.1155/2014/469298
Publisher: Hindawi Publishing Corporation
Keywords: Molecular characterization; Recombinant managanese; Superoxide dismutase; Lactococcus lactis M4
Depositing User: Nurul Ainie Mokhtar
Date Deposited: 16 Dec 2015 01:41
Last Modified: 16 Dec 2015 01:41
Altmetrics: http://www.altmetric.com/details.php?domain=psasir.upm.edu.my&doi=10.1155/2014/469298
URI: http://psasir.upm.edu.my/id/eprint/34572
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