Cloning of pdh Genes Encoding Subunits of Pyruvate Dehydrogenase Complex from Lactobacillus reuteri ATCC 55739

  • Nam, Su-Jin (Division of Applied Life Science, Graduate School, Gyeongsang National University) ;
  • Park, Jae-Yong (Division of Applied Life Science, Graduate School, Gyeongsang National University) ;
  • Kim, Jung-Kon (Division of Applied Life Science, Graduate School, Gyeongsang National University) ;
  • Ha, Yeong-Lae (Division of Applied Life Science, Graduate School, Gyeongsang National University) ;
  • Yun, Han-Dae (Division of Applied Life Science, Graduate School, Gyeongsang National University) ;
  • Kim, Jeong-Hwan (Institute of Agriculture and Life Science, Gyeongsang National University)
  • 발행 : 2004.02.01

초록

A 2-D gel protein analysis of Lactobacillus reuteri ATCC 55739 produced spots corresponding to subunits of the pyruvate dehydrogenase complex, as identified by N-terminal protein sequencing. Oligonucleotide probes specific for the subunits of the pyruvate dehydrogenase complex were synthesized ,md used to screen a L. reuteri genomic library to clone the structural genes. Two positive clones were isolated and identified as having the same 2.2 kb insert. A pdhB encoding the $\beta$-subunit of El subunit (pyruvate dehydrogenase component) of the pyruvate dehydrogenase complex was located in the middle of the insert. Furthermore, a 5' truncated pdhA encoding the $\alpha$-subunit of the E1 subunit and a 3' truncated pdhC encoding the E2 subunit (dihydrolipoamide acetyltransferase) were also located upstream and downstream of the pdhB, respectively.

키워드

참고문헌

  1. Adler, L. A. and S. Arvidson. 1988. Cloning and expression in Escherichia coli of genes encoding a multiprotein complex involved in secretion of proteins from Staphylococcus aureus. J. Bacteriol. 170: 5337-5343
  2. Bolotin, A., P. Wincker, S. Mauger, O. Jaillon, K. Malarme, J. Weissenbach, S. D. Ehrlich, and A. Sorokin. 2001. The complete genome sequence of the lactic acid bacterium Lactococcus lactis ssp. lactis IL1403. Genome Res. 11: 731-753
  3. Gottschalk, G. 1986. Bacterial Metabolism, pp. 18-20. 2$^nd$ ed. Springer-Verlag, New York, U.S.A
  4. Hammes, W., N. Weiss, and W. Holzapfel. 1992. The genera Lactobacillus and Carnobacterium, pp. 1535-1594. In Balows, A., H. G. Truper, M. Dworkin, W. Harder, and K.-H. Schleifer. (eds.), The Prokaryotes, vol II, Springer- Verlag, New York. U.S.A
  5. Harris, R. A., B. Huang, and P. Wu. 2001. Control of pyruvate dehydrogenase kinase gene expression. Adv. Enzyme Regul. 41: 269-288
  6. Hawkins, C. F., A. Borges, and R. N. Perham. 1990. Cloning and sequence analysis of the genes encoding the alpha and beta subunits of the E1 component of the pyruvate dehydrogeanse multienzyme complex of Bacillus stearothermophilus. Eur. J. Biochem. 191: 337-346
  7. Hemila, H., A. Palva, L. Paulin, S. Arvidson, and I. Palva. 1990. Secretory S complex of Bacillus subtilis: Sequence analysis and identity to pyruvate dehydrogenase. J. Bacteriol. 172: 5052-5063
  8. Hong, I.-P., H.-K. Jang, S.-Y. Lee, and S.-G. Choi. 2003. Cloning and characterization of a bifunctional cellulosechitosanase gene from Bacillus licheniformis NBL 420. J. Microbiol. Biotechnol. 13: 35-42
  9. Kim, T. Y., R. J. Park, H. C. Chang, D. K. Chung, J. H. Lee, H. J. Lee, and J. H. Kim. 2000. Cloning and characterization of the Lactococcus lactis ssp. lactis ATCC 7962 ptsHI operon. J. Microbiol. Biotechnol. 10: 829-835
  10. Liu, S., J. C. Baker, and R. E. Roche. 1995. Binding of the pyruvate dehydrogenase kinase to recombinant constructs containing the inner lipoyl domain of the dihydrolipoyl acetyltransferase component. J. Biol. Chem. 270: 793-800
  11. Matteri, A., A. de Kok, and R. N. Perham. 1992. The pyruvate dehydrogenase multienzyme complex. Curr. Opin. Struct. Biol. 2: 877-887
  12. Park, J.-Y., S.-J. Park, S. J. Nam, Y.-L. Ha, and J. H. Kim. 2002. Cloning and characterization of the L-lactate dehydrogenase gene (ldhL) from Lactobacillus reuteri ATCC 55739. J. Microbiol. Biotechnol. 12: 716-721
  13. Park, R. J., K.-H. Lee, S. J. Kim, J. Y. Park, S. J. Nam, H. D. Yun, H. J. Lee, H. C. Chang, D. K. Chung, J. H. Lee, Y. H. Park, and J. H. Kim. 2002. Isolation of Lactococcus lactis strain with b-galactosidase activity from kimchi and cloning of lacZ gene from the isolated strain. J. Microbiol. Biotechnol. 12: 157-161
  14. Ravindran, S., G. A. Radke, J. R. Guest, and T. E. Roche. 1996. Lipoyl domain based mechanism for the integrated feedback control of the pyruvate dehydrogenase complex by enhancement of pyruvate dehydrogenase kinase activity. J. Biol. Chem. 271: 653-662
  15. Sambrook, J. and D. B. Russel. 2001. Molecular Cloning, 3$^rd$ edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, U.S.A
  16. Wallbrandt, P., V. Tegman, B. H. Jonsson, and A. Wieslander. 1992. Identification and analysis of the genes coding for the putative pyruvate dehydrogenase enzyme complex in Acholeplasma laidlawii. J. Bacteriol. 174: 1388-1396