1887

Abstract

The use of a lysine-overproducing strain of in food or feed fermentations may lead to the production of lysine-rich products. The availability of functional genes and information on the regulation of lysine biosynthesis are required to develop a lysine-overproducing strain. The genome sequence of revealed putative lysine biosynthetic genes, some of which may produce isozymes. This study examined the functionality of the genes and the regulation of the first four enzymes of lysine biosynthesis, together with homoserine dehydrogenase, in . The genes were expressed in , and the regulation of the enzymes was studied in cell extracts of both recombinant and . Among seven lysine biosynthetic genes studied (aspartokinase genes, and ; aspartate semialdehyde dehydrogenase genes, and ; dihydrodipicolinate synthase genes, and ; and the dihydrodipicolinate reductase gene, ) plus two homoserine dehydrogenase genes ( and ), the products of six genes, i.e. , , , , and , showed obvious enzyme activities . The product of one of the homoserine dehydrogenase genes, , exhibited both homoserine dehydrogenase and aspartokinase activities. However, the aspartokinase activity was mainly due to ThrA2 and was inhibited by -lysine and repressed by -threonine, and the homoserine dehydrogenase activity was mainly due to Hom2 and was inhibited by -threonine. The aspartate semialdehyde dehydrogenase, dihydrodipicolinate synthase and dihydrodipicolinate reductase were not regulated by the end-products of the pathway.

Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.28092-0
2006-01-01
2024-04-19
Loading full text...

Full text loading...

/deliver/fulltext/micro/152/1/105.html?itemId=/content/journal/micro/10.1099/mic.0.28092-0&mimeType=html&fmt=ahah

References

  1. Adebawo O. O, Ruiz-Barba J. L, Warner P. J, Oguntimein G. B. O. 1997; Regulation of aspartokinase in Lactobacillus plantarum . J Appl Microbiol 82:191–196 [CrossRef]
    [Google Scholar]
  2. Barlett A. T. M, White P. J. 1986; Regulation of the enzymes of lysine biosynthesis in Bacillus sphaericus NCTC 9602 during vegetative growth. J Gen Microbiol 132:3169–3177
    [Google Scholar]
  3. Barnes I. J, Bondi A, Moat A. G. 1969; Biochemical characterization of lysine auxotrophs of Staphylococcus aureus . J Bacteriol 99:169–174
    [Google Scholar]
  4. Black S. 1962; Conversion of aspartic acid to homoserine. Methods Enzymol 5:820–827
    [Google Scholar]
  5. Black S, Wright N. G. 1955a; Aspartic β -semialdehyde dehydrogenase and aspartic β -semialdehyde. J Biol Chem 213:39–50
    [Google Scholar]
  6. Black S, Wright N. G. 1955b; β -Aspartokinase and β -aspartyl phosphate. J Biol Chem 213:27–38
    [Google Scholar]
  7. Bradford M. M. 1976; A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254 [CrossRef]
    [Google Scholar]
  8. Cohen G. N, Patte J. C. 1963; Some aspects of the regulation of amino acid biosynthesis in a branched pathway. Cold Spring Harbor Symp Quant Biol 27:513–516
    [Google Scholar]
  9. Cremer J, Treptow C, Eggeling L, Sahm H. 1988; Regulation of enzymes of lysine biosynthesis in Corynebacterium glutamicum . J Gen Microbiol 134:3221–3229
    [Google Scholar]
  10. Dower W. J, Miller J. F, Ragsdale C. W. 1988; High efficiency transformation of E. coli by high voltage electroporation. Nucleic Acids Res 16:6127–6145 [CrossRef]
    [Google Scholar]
  11. Eggeling L, Oberle S, Sahm H. 1998; Improved l-lysine yield with Corynebacterium glutamicum : use of dapA resulting in increased flux combined with growth limitation. Appl Microbiol Biotechnol 49:24–30 [CrossRef]
    [Google Scholar]
  12. Graves L. M, Switzer R. L. 1990; Aspartokinase III, a new isozyme in Bacillus subtilis 168. J Bacteriol 172:218–223
    [Google Scholar]
  13. Hegeman G. D, Cohen G. N, Morgan R. 1970; Aspartic semialdehyde dehydrogenase. Methods Enzymol 17A:708–713
    [Google Scholar]
  14. Kindler S. H, Gilvarg C. 1960; N -Succinyl-l- α , ε -diaminopimelic acid deacylase. J Biol Chem 235:3532–3535
    [Google Scholar]
  15. Kleerebezem M, Boekhorst J, van Kranenburg R et al. 2003; Complete genome sequence of Lactobacillus plantarum WCFS1. Proc Natl Acad Sci U S A 100:1990–1995 [CrossRef]
    [Google Scholar]
  16. Misono H, Togawa H, Yamamoto T, Soda K. 1979; meso - α , ε -Diaminopimelate d-dehydrogenase: distribution and the reaction product. J Bacteriol 137:22–27
    [Google Scholar]
  17. Morishita T, Deguchi Y, Yajima M, Sakurai T, Yura T. 1981; Multiple nutritional requirements of lactobacilli: genetic lesions affecting amino acid biosynthetic pathways. J Bacteriol 148:64–71
    [Google Scholar]
  18. Nakayama K, Tanaka H, Hagino H, Kinoshita S. 1966; Studies on lysine fermentation. Part V. Concerted feedback inhibition of aspartokinase and the absence of lysine inhibition on aspartic semialdehyde-pyruvate condensation in Micrococcus glutamicus . Agric Biol Chem 30:611–616 [CrossRef]
    [Google Scholar]
  19. Patte J. C, Le Bras G, Cohen G. N. 1967; Regulation by methionine of the synthesis of a third aspartokinase and of a second homoserine dehydrogenase in Escherichia coli K12. Biochim Biophys Acta 136:245–257 [CrossRef]
    [Google Scholar]
  20. Rosner A, Paulus H. 1971; Regulation of aspartokinase in Bacillus subtilis . J Biol Chem 246:2965–2971
    [Google Scholar]
  21. Selli A, Crociani F, Di Gioia D, Fava F, Crisetig G, Matteuzzi D. 1994; Regulation of dihydrodipicolinate synthase and diaminopimelate decarboxylase activity in Bacillus stearothermophilus . Ital J Biochem 43:29–35
    [Google Scholar]
  22. Sundharadas G, Gilvarg C. 1967; Biosynthesis of α , ε -diaminopimelic acid in Bacillus megaterium . J Biol Chem 242:3983–3988
    [Google Scholar]
  23. Theze J, Margarita D, Cohen G. N, Borne F, Patte J. C. 1974; Mapping of the structural genes of the three aspartokinases and of the two homoserine dehydrogenases of Escherichia coli K-12. J Bacteriol 117:133–143
    [Google Scholar]
  24. Tosaka O, Takinami K. 1978; Pathway and regulation of lysine biosynthesis in Brevibacterium lactofermentum . Agric Biol Chem 42:95–100 [CrossRef]
    [Google Scholar]
  25. Umbarger H. E. 1978; Amino acid biosynthesis and its regulation. Annu Rev Biochem 47:533–606 [CrossRef]
    [Google Scholar]
  26. Vrljic M, Kronemeyer W, Sahm H, Eggeling L. 1995; Unbalance of l-lysine flux in Corynebacterium glutamicum and its use for the isolation of excretion-defective mutants. J Bacteriol 177:4021–4027
    [Google Scholar]
  27. Weinberger S, Gilvarg C. 1970; Bacterial distribution of the use of succinyl and acetyl blocking groups in diaminopimelic acid biosynthesis. J Bacteriol 101:323–324
    [Google Scholar]
  28. White P. J. 1976; The regulation of diaminopimelate decarboxylase activity in Escherichia coli strain w. J Gen Microbiol 96:51–62 [CrossRef]
    [Google Scholar]
  29. White P. J. 1983; The essential role of diaminopimelate dehydrogenase in the biosynthesis of lysine by Bacillus sphaericus . J Gen Microbiol 129:739–749
    [Google Scholar]
  30. Yamakura F, Ikeda Y, Kimura K, Sasakawa T. 1974; Partial purification and some properties of pyruvate-aspartic semialdehyde condensing enzyme from sporulating Bacillus subtilis . J Biochem 76:611–621
    [Google Scholar]
  31. Yugari Y, Gilvarg C. 1962; Coordinated end-product inhibition in lysine synthesis in Escherichia coli . Biochim Biophys Acta 62:612–614 [CrossRef]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.28092-0
Loading
/content/journal/micro/10.1099/mic.0.28092-0
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error