Food-grade expression of human glutathione S-transferase and Cu/Zn superoxide dismutase in Lactococcus lactis.
A food-grade gene expression system in Lactococcus lactis was established by the combination of a vector containing the lacF gene as the selection marker and a strain WZ103 carrying an in-frame deletion of this gene in the chromosome as the host. The human glutathione S-transferase A1-1 (hGSTA1) and Cu/Zn superoxide dismutase (hSOD) genes were respectively cloned into a food-grade vector under the control of the lactococcal inducible promoter P(lacA). The resulting expression plasmids were separately introduced into the lactose-deficient (Lac(-)) host, and the lactose-utilizing (Lac(+)) transformants were directly selected on a chemically defined medium, using lactose as the sole carbon source. The successful food-grade expression of hGSTA1 and hSOD in the L. lactis WZ103 transformed with these plasmids were analyzed by Western blotting and enzymatic activity assay, respectively.
Xiang
Hua
H
Institute of Microbiology, Chinese Academy of Sciences, Beijing 100080, China.
Wei
Wenzhong
W
Tan
Huarong
H
D003001:Cloning, Molecular; D005516:Food Microbiology; D030841:Food, Genetically Modified; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology; D005982:Glutathione Transferase / Q000096:biosynthesis* / Q000235:genetics*; D006801:Humans; D007763:Lac Operon; D013294:Lactococcus lactis / Q000201:enzymology* / Q000235:genetics*; D015202:Protein Engineering / Q000379:methods; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D013482:Superoxide Dismutase / Q000096:biosynthesis* / Q000235:genetics*
D011994:Recombinant Proteins; D013482:Superoxide Dismutase; D005982:Glutathione Transferase