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Cloning and Expression of Manganese Superoxide Dismutase Gene by Food-Grade Vector in Lactococcus lactis

MA Hongdan1, ZHANG Liyuan1, ZHAO Handan1, XU Dandan1, QU Jing2, GUAN Shuyan1,*, WANG Piwu2,*   

  1. 1. College of Life Sciences, Jilin Agricultural University, Changchun 130118, China;
    2. College of Agriculture, Jilin Agricultural University, Changchun 130118, China
  • Online:2015-01-15 Published:2015-01-16

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Abstract:

Open reading frame of manganese superoxide dismutase (MnSOD) from soybean was cloned into the plasmid
pNZ8149 and plasmid pNZS was reformed to construct the food-grade vector. The recombinant plasmids were then
separately transformed into L. lactis NZ3900 by electronic perforation, and the growth ability of the transformants was
detected in Elliker medium. The recombinant plasmids named pNZ-SOD and pNZS-SOD were thus constructed successfully
after PCR amplification, ligation and identification. The expression of L. lactis NZ3900/pNZ-SOD and L. lactis NZ3900/
pNZS-SOD was induced by nisin and their expressed products were analyzed by sodium dodecyl sulfate-polyacrylamide gel
electrophoresis (SDS-PAGE). The SOD enzymatic activities of the transformants L. lactis NZ3900/pNZ-SOD and L. lactis
NZ3900/pNZ-SOD were determined by the inhibition of nitrotetrazolium. The SOD enzymatic activity of the transformant
L. lactis NZ3900/pNZS-SOD (with two thirds of extracellular SOD) was 1.6 times higher than that of the transformant
L. lactis NZ3900/pNZS-SOD and 13.5 times higher than that of the parent strain. Hence, the expression of MnSOD was
enhanced in L. lactis NZ3900/pNZS.

Key words: manganese superoxide dismutase (MnSOD), food-grade vector, Lactococcus lactis

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