Cloning and Expression of the UDP-Galactose-4-Epimerase Gene (galE) Constituting the gal/lac Operon of Lactococcus lactis ssp. lactis ATCC7962

  • Lee, Jung-Min, Choi, Jae-Yeon (Department of Food Science and Technology, Seoul National University) ;
  • Lee, Jong-Hoon (Department of Foods and Biotechnology, Kyonggi University) ;
  • Chang, Hae-Choon (Department of Foods and Biotechnology, Kyonggi University) ;
  • Chung, Dae-Kyun (Department of Food and Nutrition, Chosun University) ;
  • Kim, Jeong-Hwan (Institute and Department of Genetic Engineering, KyungHee University) ;
  • Lee, Hyong-Joo (Department of Food Science and Technology, Gyeongsang National University) ;
  • Published : 1999.08.01

Abstract

The gene (galE) encoding UDP-galactose-4-epimerase, operative in the galactose metabolic pathway, was cloned together with the $\beta$-galactosidase gene (lacZ) from Lactococcus lactis ssp. lactis ATCC7962 (L. lactis 7962). galE was found to have a length of 981 bps and encoded a protein with a molecular mass of 36,209 Da. The deduced amino acid sequence showed a homology with GalE proteins from several other microorganisms. A Northern analysis demonstrated that galE was constitutively expressed by its own promoter. When galactose or lactose was added into medium, the galE transcription was induced by several upstream promoters. The structure of the gal/lac operon of L. lactis 7962 was partially characterized and the gene order around galE was galT-lacA-lacZ-galE-orfX.

Keywords

References

  1. J. Bacteriol. v.170 Gene organization and structure for the Streptomyces lividans gal operon Adams, C. W.;J. A. Fornwald;F. J. Schmidt;M. Rosenberg;M. E. Brawner
  2. Esherichia coli and Salmonella typhimurium: Cellular and Molecular Biology v.2 Adhya, S.;F. C. Neidhardt(ed.);J. L. Ingraham(ed.);K. B. Low(ed.);B. Magasanik(ed.);M. Schaechter(ed.);H. E. Umbarger(ed.)
  3. Proteins: Structure, Function, and Genetics v.12 The molecular structure of UDP-galactose-4-epimerase from Escherichia coli determined at 2.5 Å resolution Bauer, A. J.;I. Rayment;P. A. Frey;M. Holden
  4. Nucl. Acids Res. v.7 A rapid alkaline extraction procedure for screening recombinant plasmid DNA Bimboim, H. C.;J. Daly
  5. J. Bacteriol. v.117 Lactose and D-galactose metabolism in group N Streptococci: Presence of enzymes for both the D-galactose-l-phosphate and D-galactose-6-phosphate pathways Bissett, D. L.;R. L. Anderson
  6. J. Microbiol. Biotechnol. v.6 Molecular cloning of a β-galactosidase gene from Lactococcus lactis subsp. lactis 7962 Chang, H.-C.;Y.-D. Choi;H.-J. Lee
  7. Mol. Microbiol. v.5 The Rhizobium meliloti exoZ1 exoB fragment of megaplasmid 2: exoB functions as a UDP-glucose-4-epimerase and ExoZ shows homology to NodX of Rhizobium leguminosarum biovar. viciae strain TOM Buendia, A. M.;B. Enenkel;R. Koplin;K. Niehaus;W. Arnold;A. Puhler
  8. J. Bacteriol. v.170 Characterization of a gene cluster for exopolysaccharide biosynthesis and virulence in Erwinia stewartii Dolph, P. J.;D. R. Majerczak;D. L. Colpin
  9. Biotechnol. Lett. v.16 The gene (lacA) encoding galactoside acetyltransferase from Lactococcus lactis Griffin, H. G.;M. J. Gasson
  10. J. Microbiol. Biotechnol v.9 Effect of temperature and carbon source on the expression of β-galactosidase gene of Lactococcus lactis ssp. lactis ATCC7962 Kim, T. Y.;J. M. Lee
  11. Probiotic Researches on Lactic Acid Bacteria. Ann. Meet. Kor. Soc. Appl. Microbiol. Characterization of the gal/lac of Lactococcus lactis ssp. lactis ATCC7962 Lee, H. J.;J. H. Kim;H. C. Chang;D. K. Chung
  12. Biotechnol. Lett. v.19 Cloning and nucleotide sequence of the β-galactosidase gene from Lactococcus lactis ssp. lactis ATCC7962 Lee, J. M.;D. K. Chung;J. H. Park;W. K. Lee;H. C. Chang;J. H. Kim;H. J. Lee
  13. Nature v.227 Clcavage of structural proteins during the assembly of the head of bacteriophage T4 Laemmli, U. K.
  14. Nucl. Acids Res. v.14 Nucleotide sequences of the galE gene and the galT gene of Escherichia coli Lemaire, H. G.;B. Muller-Hill
  15. J. Bacteriol. v.173 Lactose metabolism in Lactobacillus bulgaricus: Analysis of the primary structure and expression of the genes involved Leong-Morgenthalcr, P.;M. C. Zwahlen;H. Hottinger
  16. J. Biol. Chem. v.261 UDP-glucose-4-epimerase from Saccharomyces fragilis. Presence of an essential arginine residue at the substrate-binding site of the enzyme Mukherji, S.;A. Bhaduri
  17. J. Bacteriol. v.171 Lactose transport system of Streptococcus thermophilus: A hybrid protein with homology to the melibiose carrier and enzyme Ⅲ of phosphoenolpyruvate-dapendent phosphotransferase systems Poolman, B.;T. J. Royer;S. E. Mainzer;B. F. Schmit
  18. J. Bacteriol. v.172 Carbohydrate utilization in Streptococcus thermophilus: Characterization of the genes for aldose-1-epimerase(mularotase) and UDP-glucose-4-epimerase Poolman, B.;T. J. Royer;S. E. Mainzer;B. F. Schmidt
  19. Biochem. Biophys. Acta v.457 The bacterial phosphoenolpyruvate: Sugar phosphotransferase system Postman, P.;S. Roseman
  20. Molecular Cloning: A Laboratory Manual Sambrook, J.;E. F. Fritsch;T. Maniatis
  21. J. Bacteriol. v.84 Cell-wall composition and the grouping antigens of Streptoccocci Slade, H. D.;W. C. Slamp
  22. J. Bacteriol. v.144 Galactose fermentation by Streptococcus lactis and Strptococcus cremoris: Pathways, products, and regulation Thomas, T. D.;K. W. Turner;V. L. Crow
  23. Nucl. Acids Res. v.16 Nucleotide sequence of the galactose gene cluster of Kluyveromyce lactis Webster, T. D.;R. C. Dickson
  24. Nucl. Acids Res. v.16 The organization and transcription of the galactose gene of Kluyveromyce lactis Webster, T. D.;R. C. Dickson