1887

Abstract

In -lysine is synthesized simultaneously via the succinylase and dehydrogenase variant of the diaminopimelate pathway. Starting from a strain with a disrupted dehydrogenase gene, three different-sized DNA fragments were isolated which complemented defective mutants in the succinylase pathway. Enzyme studies revealed that in one case the dehydrogenase gene had apparently been reconstituted in the heterologous host. The two other fragments resulted in desuccinylase activity; one of them additionally in succinylase activity. However, the physical analysis showed that structural changes had taken place in all fragments. Using a probe derived from one of the fragments we isolated a 3.4 kb Hl DNA fragment without selective pressure (by colony hybridization). This was structurally intact and proved functionally to result in tenfold desuccinylase overexpression. The nucleotide sequence of a 1966 bp fragment revealed the presence of one truncated open reading frame of unknown function and that of encoding -succinyl diaminopimelate desuccinylase (EC 3.5.1.18). The deduced amino acid sequence of the gene product shares 23% identical residues with that from The gene now available is the first gene from the succinylase branch of lysine synthesis of this biotechnologically important organism.

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1994-12-01
2024-12-07
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References

  1. Abe S., Takayama K., Kinoshita S. Taxonomical studies on glutamic acid-producing bacteria. J Gen Appl Microbiol 1967; 13:279–301
    [Google Scholar]
  2. Baril C., Richaud C., Fourne G., Baranton G., Saint Girons I. Cloning of dapD, aroD and asd of Leptospira interrogans serovar icterohaemorrhagiae, and nucleotide sequence of the asd gene. J Gen Microbiol 1992; 138:47–53
    [Google Scholar]
  3. Bouvier J., Richaud C., Higgins W., Bogler O., Stragier P. Cloning, characterization, and expression of the dapE gene of Escherichia coli. J Bacteriol 1992; 174:5265–5271
    [Google Scholar]
  4. Cordes C., Mockel B., Eggeling L., Sahm H. Cloning, organization and functional analysis of ilvA, ilvB and ilvC genes from Corynebacterium glutamicum. Gene 1992; 112:113–116
    [Google Scholar]
  5. Cremer J., Treptow C., Eggeling L., Sahm H. Regulation of enzymes of lysine biosynthesis in Corynebacterium glutamicum. J Gen Microbiol 1988; 134:3221–3229
    [Google Scholar]
  6. Cremer J., Eggeling L., Sahm H. Cloning the dap A dapB cluster of the lysine-secreting bacterium Corynebacterium glutamicum. Mol & Gen Genet 1990; 220:478–480
    [Google Scholar]
  7. Cremer J., Eggeling L., Sahm H. Control of the lysine biosynthetic sequence in Corynebacterium glutamicum as analyzed by overexpression of the individual corresponding genes. Appl Environ Microbiol 1991; 57:1746–1752
    [Google Scholar]
  8. Denich K., O'Hanley P., Lalonde G. Cloning and sequence analysis of the DapD gene from Actinobacillus pleuro-pneumoniae. EMBL Database 1991X63201
    [Google Scholar]
  9. Eggeling L. Biology of L-lysine overproduction by Corynebacterium glutamicum. Amino Acids 1994; 6:261–272
    [Google Scholar]
  10. Eikmanns B. Identification, sequence analysis, and expression of a Corynebacterium glutamicum gene cluster encoding the three glycolytic enzymes glyceraldehyde-3-phosphate dehydrogenase, 3-phosphoglycerate kinase, and triosephosphate isomerase. J Bacteriol 1992; 174:6067–6086
    [Google Scholar]
  11. Gilvarg C. JV-Succinyl-L-diaminopimelic acid. J Biol Chem 1959; 234:2955–2959
    [Google Scholar]
  12. Gornall A.G., Bardawill C.J., David M.M. Determination of serum proteins by means of the biuret reaction. J Biol Chem 1949; 77:751–766
    [Google Scholar]
  13. Gough J.A., Murray N.E. Sequence diversity among related genes for recognition of specific targets in DNA molecules. J Mol Biol 1983; 166:1–19
    [Google Scholar]
  14. Hanahan D. Techniques for transformation of E. coli. In DNA Cloning: a Practical Approach 1985 Edited by Glover D.M. Oxford: IRL Press; 1 pp 109–136
    [Google Scholar]
  15. Hohn B., Collins J. A small cosmid for efficient cloning of large DNA fragments. Gene 1980; 11:291–298
    [Google Scholar]
  16. Ishino S., Muzikami T., Yamaguchi K., Katsumata R., Araki K. Cloning and sequencing of the meso-diaminopimelate-D-dehydrogenase (ddh) gene of Corynebacterium glutamicum. Agric Biol Chem 1988; 52:2903–2909
    [Google Scholar]
  17. Kalinowski J., Bachmann B., Thierbach G., Piihler A. Aspartokinase genes lysCa. and lysCp overlap and are adjacent to the aspartate β-semialdehyde dehydrogenase gene asd in Corynebacterium glutamicum. Mol & Cen Genet 1990; 224:317–324
    [Google Scholar]
  18. Lennox E.S. Transduction of linked genetic characters of the host by bacteriophage PI. Virology 1955; 1:190–206
    [Google Scholar]
  19. Liebl W., Bayerl A., Stiilner U., Schleifer K.H. High efficiency electroporation of intact Corynebacterium glutamicum cells. FEMS Microbiol Eett 1989; 65:299–304
    [Google Scholar]
  20. Menkel E., Thierbach G., Eggeling L., Sahm H. Influence of increased aspartate availability on lysine formation by a recombinant strain of Corynebacterium glutamicum and utilization of fumarate. Appl Environ Microbiol 1989; 55:684–688
    [Google Scholar]
  21. Mockel B., Eggeling L., Sahm H. Functional and structural analyses of threonine dehydratase from Corynebacterium glutamicum. J Bacteriol 1992; 174:8065–8072
    [Google Scholar]
  22. Moran C.P., Lang N., Le Grice S.F.J., Lee G., Stephens M., Sonenshein A.L., Pero J., Losick R. Nucleotide sequences that signal the initiation of transcription and translation in Bacillus subtilis. Mol & Gen Genet 1982; 186:339–346
    [Google Scholar]
  23. Pätek M., Krumbach K., Eggeling L., Sahm H. Leucine synthesis in Corynebacterium glutamicum : enzyme activities, structure of leuA, and effect of leu A inactivation on lysine synthesis. Appl Environ Microbiol 1994; 60:133–140
    [Google Scholar]
  24. Richaud F., Richaud C., Haziza C., Patte J.-C. Isolement et purification de gènes d'Escherichia coli Kl2 impliqués dans la biosynthèse de la lysine. C R Acad Sei Ser III 1981; 293:507–512
    [Google Scholar]
  25. Richaud C., Richaud F., Martin C., Haziza C., Patte J.-C. Regulation of expression and nucleotide sequence of the Escherichia coli dapD gene. J Biol Chem 1984; 259:14824–14828
    [Google Scholar]
  26. Richaud C., Higgins W., Mengin-Lecreulx D., Stragier P. Molecular cloning, characterization, and chromosomal localization of dapF, the Escherichia coli gene for diaminopimelate epimerase. J Bacteriol 1987; 169:1454–1459
    [Google Scholar]
  27. Sanger F., Nicklen S., Coulson A.R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sei USA 1977; 74:5463–5467
    [Google Scholar]
  28. Schrumpf B., Schwarzer A., Kalinowski J., Pühler A., Eggeling L., Sahm H. A functionally split pathway for lysine synthesis in Corynebacterium glutamicum. J Bacteriol 1991; 173:4510–4516
    [Google Scholar]
  29. Schwinde J.W., Thum-Schmitz N., Eikmanns B.J., Sahm H. Transcriptional analysis of the gap-pgk-tpi-ppc gene cluster of Corynebacterium glutamicum. J Bacteriol 1993; 175:3905–3908
    [Google Scholar]
  30. Sonntag K., Eggeling L., De Graaf A.A., Sahm H. Flux partitioning in the split pathway of lysine synthesis in Corynebacterium glutamicum. Quantification by 13C-and 'H-NMR spectroscopy. Eur J Biochem 1993; 213:1325–1331
    [Google Scholar]
  31. Southern E.M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 1975; 98:503–517
    [Google Scholar]
  32. Weinberger S., Gilvarg C. Bacterial distribution of the use of succinyl and acetyl blocking groups in diaminopimelic acid biosynthesis. J Bacteriol 1970; 101:323–324
    [Google Scholar]
  33. White P.J. The essential role of diaminopimelate dehydrogenase in the biosynthesis of lysine by Bacillus sphaericus. J Gen Microbiol 1983; 129:739–749
    [Google Scholar]
  34. Wu B., Georgopoulos C., Ang D. The essential Escherichia coli msgB gene, a multicopy suppressor of a temperaturesensitive allele of the heat shock gene grpE, is identical to dapE. J Bacteriol 1992; 174:5258–5264
    [Google Scholar]
  35. Yanisch-Perron C., Vieira J., Messing J. Improved Ml 3 phage cloning vectors and host strains: nucleotide sequences of the M13mpl8 and pUC19 vectors. Gene 1985; 33:103–109
    [Google Scholar]
  36. Yeh P., Sicard A.M., Sinskey A.J. General organization of the genes specifically involved in the diaminopimelate-lysine biosynthetic pathway of Corynbacterium glutamicum. Mol & Gen Genet 1988; 212:105–111
    [Google Scholar]
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