1887

Abstract

Lipoic acid () and 2-oxoglutarate dehydrogenase () mutants of 12 exhibit a requirement for exogenous succinate during aerobic growth on glucose minimal medium. Reversion studies have shown that this requirement can be suppressed by -linked mutations which inactivate succinate dehydrogenase. Biochemical and genetic studies confirmed that the succinate dehydrogenase gene () is affected and that suppression is mediated by the same intergenic and indirect mechanism that generates succinate independence in partial revertants of lipoamide dehydrogenase mutants ( Creaghan & Guest, 1977 ).

A series of isogenic strains containing all combinations of mutations affecting 2-oxoglutarate dehydrogenase (), succinate dehydrogenase (), isocitrate lyase () and fumarate reductase () in a background lacking succinate semialdehyde dehydrogenase, was constructed to assess the importance of these enzymes as sources of endogenous succinate (succinyl-CoA) during aerobic and anaerobic growth on glucose. Only strains combining a deficiency in 2-oxoglutarate dehydrogenase with the presence of an active succinate dehydrogenase required succinate for aerobic growth. In all mutants, including the triple mutant (), the succinate requirement was suppressed by inactivating succinate dehydrogenase. The aerobic growth rates of succinate-independent strains were most affected by lack of isocitrate lyase but only two mutants ( and ) grew faster with added succinate: the growth yields were lowered by deficiencies in isocitrate lyase and also succinate dehydrogenase. It is concluded that very little succinate is needed for biosynthesis during aerobic growth on glucose and the requirement for relatively high concentrations of succinate (2 m) by mutants lacking 2-oxoglutarate dehydrogenase or related functions stems from the presence of active succinate dehydrogenase. Anaerobically, either isocitrate lyase or fumarate reductase is essential for succinate-independent growth on glucose.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-107-1-1
1978-07-01
2024-04-19
Loading full text...

Full text loading...

/deliver/fulltext/micro/107/1/mic-107-1-1.html?itemId=/content/journal/micro/10.1099/00221287-107-1-1&mimeType=html&fmt=ahah

References

  1. Amarasingham C. R., Davis B. D. 1965; Regulation of α-ketoglutarate dehydrogenase formation in Escherichia coli . Journal of Biological Chemistry 240:3664–3668
    [Google Scholar]
  2. Bachmann B. J., Low K. B., Taylor A. L. 1976; Recalibrated linkage map of Escherichia coli k12. Bacteriological Reviews 40:116–167
    [Google Scholar]
  3. Brice C. B., Kornberg H. L. 1968; Genetic control of isocitrate lyase activity in Escherichia coli . Journal of Bacteriology 96:2185–2186
    [Google Scholar]
  4. Cox G. B., Newton N. A., Gibson F., Snoswell A. M., Hamilton J. A. 1970; The function of ubiquinone in Escherichia coli . Biochemical Journal 117:551–562
    [Google Scholar]
  5. Creaghan I. T., Guest J. R. 1972; Amber mutants of the α-ketoglutarate dehydrogenase gene of Escherichia coli k12. Journal of General Microbiology 71:207–220
    [Google Scholar]
  6. Creaghan I. T., Guest J. R. 1977; Suppression of the succinate requirement of lipoamide dehydrogenase mutants of Escherichia coli by mutations affecting succinate dehydrogenase activity. Journal of General Microbiology 102:183–194
    [Google Scholar]
  7. Dover S., Halpern Y. S. 1972; Utilisation of y-aminobutyric acid as the sole carbon and nitrogen source by Escherichia coli k12 mutants. Journal of Bacteriology 109:835–843
    [Google Scholar]
  8. Guest J. R. 1974; Gene-protein relationships of the α-keto acid dehydrogenase complexes of Escherichia coli k12 : chromosomal location of the lipoamide dehydrogenase gene. Journal of General Microbiology 80:523–532
    [Google Scholar]
  9. Guest J. R., Creaghan I. T. 1973; Gene-protein relationships of the α-keto acid dehydrogenase complexes of Escherichia coli k12: isolation and characterization of lipoamide dehydrogenase mutants. Journal of General Microbiology 75:197–210
    [Google Scholar]
  10. Guest J. R., Creaghan I. T. 1974; Further studies with lipoamide dehydrogenase mutants of Escherichia colik12. Journal of General Microbiology 81:237–245
    [Google Scholar]
  11. Herbert A. A., Guest J. R. 1968; Biochemical and genetic studies with lysine + methionine mutants of Escherichia coli: lipoic acid and α-ketoglutarate dehydrogenase-less mutants. Journal of General Microbiology 53:363–381
    [Google Scholar]
  12. Herbert A. A., Guest J. R. 1969; Studies on α-keto acid dehydrogenase mutants of Escherichia coli . Molecular and General Genetics 105:182–190
    [Google Scholar]
  13. Lambden P. R., Guest J. R. 1976; Mutants of Escherichia coli k12 unable to use fumarate as an anaerobic electron acceptor. Journal of General Microbiology 97:145–160
    [Google Scholar]
  14. Lennox E. S. 1955; Transduction of linked genetic characters of the host by bacteriophage P1.. Virology 1:190–206
    [Google Scholar]
  15. Porra R. J., Jones O. T. G. 1963; Studies on ferrochelatase. 1. Assay and properties of ferro-chelatase from a pig-liver mitochondrial extract. Biochemical Journal 87:181–192
    [Google Scholar]
  16. Spencer M. E., Guest J. R. 1973; Isolation and properties of fumarate reductase mutants of Escherichia coli . Journal of Bacteriology 114:563–570
    [Google Scholar]
  17. Spencer M. E., Lebeter V. M., Guest J. R. 1976; Location of the aspartase gene (aspA) on the linkage map of Escherichia coli k12. Journal of General Microbiology 97:73–82
    [Google Scholar]
  18. Stouthamer A. H. 1973; A theoretical study on the amount of ATP required for synthesis of microbial cell material. Antonie van Leeuwenhoek 39:545–565
    [Google Scholar]
  19. Vinopal R. T., Fraenkel D. G. 1974; Pheno-typic suppression of phosphofructokinase mutations in Escherichia coli by constitutive expression of the glyoxylate shunt. Journal of Bacteriology 118:1090–1100
    [Google Scholar]
  20. Vise A. B., Lascelles J. 1967; Some properties of a mutant strain of Escherichia coli which requires lysine and methionine or lipoic acid for growth. Journal of General Microbiology 48:87–93
    [Google Scholar]
  21. Vogel H., Bonner D. M. 1956; A convenient growth medium for Escherichia coli and some other micro-organisms. Microbial Genetics Bulletin 13:43–44
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-107-1-1
Loading
/content/journal/micro/10.1099/00221287-107-1-1
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