1887

Abstract

SUMMARY: An approach has been made to the problem of how the synthesis of coenzymes is regulated. Two aspects of the problem have been studied, especially as they concern the synthesis of flavins by bacteria:

(1) How are coenzymes prevented from being synthesized as fast as amino acids or nucleic acid bases?

(2) How is coenzyme synthesis adjusted to the often changing physiological needs of bacteria?

Evidence is presented that flavins cannot inhibit the activity of enzymes in the flavin biosynthetic pathway of , but the amount of these enzymes can be made to vary by a factor of at least two. Repression might, therefore, account for the low rate of flavin synthesis. The possibility that repression rather than feedback inhibition also accounts for the low rate of synthesis of other coenzymes is discussed.

Flavin synthesis is not as precisely adjusted to the physiological needs of bacteria as are syntheses of major metabolites for the following reasons:

(1) Flavins are greatly overproduced by bacteria during exponential growth; the ratio of flavins excreted to flavins retained in the cells is between 0.8 and 8 for all strains and cultural conditions tested.

(2) Flavin synthesis is not tightly geared to growth; thus, flavin synthesis goes on uninterrupted for more than an hour when the growth rate of or is abruptly reduced from a rapid rate to zero; also growth goes on uninterrupted for over an hour when the flavin supply is abruptly cut off from rapidly growing lactic acid bacteria. Evidently the control mechanism in the flavin pathway is not very sensitive to physiological needs. This conclusion probably applies to other coenzymes as well.

Some incidental findings of interest from other points of view were:

(1) Although internal flavins can get out of , external flavins apparently cannot enter. This could account for the absence of flavinless mutants.

(2) After brief treatment with penicillin, becomes permeable to external flavins while remaining both impermeable to inulin and capable of synthesizing flavins.

(3) Less than 4 % of the intracellular flavins of are free, in a form that can be extracted with -butanol (5 %, v/v), toluene (0.05 %, v/v), cetyl trimethylammonium bromide (0.001 %, w/v) or distilled water. The remaining flavins are bound, in a form that can be extracted with trichloracetic acid (5 %, w/v). The intracellular concentration of free flavins in is estimated to be less than 4 x 10M.

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1962-06-01
2024-04-18
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References

  1. Bolton E. T., Britten R. J., Cowie D. B., Roberts R. B. 1955; Biophysics. Yearb. Carneg. Instn 54:75
    [Google Scholar]
  2. Brooke M. S., Ushiba D., Magasanik B. 1954; Some factors affecting the excretion of orotic acid by mutants of Aerobacter aerogenes. J. Bact 68:534
    [Google Scholar]
  3. Burch H. B. 1957; Fluorimetric assay of FAD, FMN and riboflavin. Methods in Enzymology III:960 Colowick S. P., and Kaplan N. O. New York: Academic Press;
    [Google Scholar]
  4. Dagley S., Johnson A. R. 1956; Appearance of amino acids and peptides in culture filtrates of micro-organisms growing in mineral salt media. Biochim. biophys. Acta 21:270
    [Google Scholar]
  5. Davis B. D. 1950; Studies on nutritionally deficient bacterial mutants isolated by means of penicillin. Experientia 6:41
    [Google Scholar]
  6. Eagle H., Agranoff B. W., Snell E. E. 1960; The biosynthesis of meso-inositol by cultured mammalian cells, and the parabiotic growth of inositol-dependent and inositol-independent strains. J. biol. Chem 235:1891
    [Google Scholar]
  7. Enari T. M. 1958; Studies on the uptake of cobalt and iron and effect on the production of riboflavin by Candida guiltiermondii. Ann. Acad. Sci. fennicae Ser. A, sec. 2 90:1
    [Google Scholar]
  8. Ennis H. L., Gorini L. 1961; Control of arginine biosynthesis in strains of Escherichia coli not repressible by arginine. J. mol. Biol 3:439
    [Google Scholar]
  9. Euler H., von & Adler E. 1934; Freies Flavin und gebundenes Flavin (Flavin-Enzym) in tierischen Organen und Flüssigkeiten und in Hefe. Arkiv. Kemi Min. Geol. B 11:28
    [Google Scholar]
  10. Gale E. F., Folkes J. B. 1953; The assimilation of amino-acids by bacteria, 15. Actions of antibiotics on nucleic acid and protein synthesis in Staphylococcus aureus. Biochem. J 53:493
    [Google Scholar]
  11. Goodwin T. W., McEvoy D. 1959; Studies on the biosynthesis of riboflavin, 5. General factors controlling flavinogenesis in the yeast Candida flareri. Biochem. J 71:742
    [Google Scholar]
  12. Goodwin T. W., Pendlington S. 1954; Studies on the biosynthesis of riboflavin. Nitrogen metabolism and flavinogenesis in Eremothecium ashbyii. Biochem. J 57:631
    [Google Scholar]
  13. Hanawalt P., Setlow R. 1960; Effect of monochromatic light on macromoleeular synthesis in Escherichia coli.. Biochim. biophys. Acta 41:283
    [Google Scholar]
  14. Imsande J. 1961; Pathway of diphosphopyridine nucleotide biosynthesis in Escherichia coli. J. biol. Chem 236:1494
    [Google Scholar]
  15. Imsande J., Pardee A. B. 1962; Regulation of pyridine nucleotide biosynthesis in Escherichia coli.. J. biol. Chem237 (in the Press)
    [Google Scholar]
  16. Kearney E. B., Englard S. 1951; The enzymatic phosphorylation of riboflavin. J. biol. Chem 193:821
    [Google Scholar]
  17. Kitay E., Snell E. E. 1948; Effect of size of inoculum on the apparent vitamin requirements of lactic acid bacteria. Proc. Soc. exp. Biol. Med 68:648
    [Google Scholar]
  18. Knight B. C. J. G. 1945; Growth factors in microbiology. Some wider aspects of nutritional studies with micro-organisms. Vitam. & Horm 3:105
    [Google Scholar]
  19. Lascelles J. 1961; Synthesis of tetrapyrroles by micro-organisms. Physiol. Rev 41:417
    [Google Scholar]
  20. Lascelles J., Woods D. D. 1952; The synthesis of ‘folic acid’ by Bacterium coli and Staphylococcus aureus and its inhibition by sulphonamides. Brit. J. exp. Path 33:288
    [Google Scholar]
  21. Lederberg J. 1950; Isolation and characterization of biochemical mutants of bacteria. Meth. med. Res 3:5
    [Google Scholar]
  22. Lowry O. H., Rosebrough N. J., Farr A. L., Randall R. J. 1951; Protein measurement with the Folin phenol reagent. J. biol. Chem 193:265
    [Google Scholar]
  23. Maas W. K. 1959a; Effect of penicillin on the uptake of amino acids in bacteria. Biochem. biophys. Res. Comm 1:13
    [Google Scholar]
  24. Maas W. K. 1959b; The biosynthesis of pantothenic acid. Congr. int. Biochem. Symp 11:161
    [Google Scholar]
  25. MacCormick D. B., Gregory M. E., Snell E. E. 1961; Pyridoxal phosphokinases I. Assay, distribution, purification and properties. J. biol. Chem 236:2076
    [Google Scholar]
  26. Maley G. F., Plaut G. W. E. 1959; The isolation, synthesis and metabolic properties of 6,7-dimethyI-8-ribityIlumazine. J. biol. Chem 234:641
    [Google Scholar]
  27. McIlwain H. 1946a; The magnitude of microbial reactions involving vitamin-like compounds. Nature, Lond. 158:898
    [Google Scholar]
  28. McIlwain H. 1946b; The metabolism and functioning of vitamin-like compounds, 2. A comparison of pantothenate metabolism by proliferating and non-proliferating bacteria. Biochem. J 40:269
    [Google Scholar]
  29. Mitchell P., Moyle J. 1956; Osmotic function and structure in bacteria. Bacterial Anatomy. Symp. Soc. gen. Microbiol 6:150
    [Google Scholar]
  30. Morris J. G. 1959; The synthesis of vitamin B6 by some mutant strains of Escherichia coli. J. gen. Microbiol 20:597
    [Google Scholar]
  31. Moyed H. S. 1960; False feedback inhibition: inhibition of tryptophan biosynthesis by 5-methyltryptophan. J. biol. Chem 235:1098
    [Google Scholar]
  32. Newton B. A. 1958; Surface-active bactericides. The Strategy of Chemotherapy. Symp. Soc. gen. Microbiol 8:62
    [Google Scholar]
  33. Nimmo-Smith R. H., Lascelles J., Woods D. D. 1948; The synthesis of ‘folic acid’ by Streptobaderium plantarum and its inhibition by sulphonamides. Brit. J. exp. Path 29:264
    [Google Scholar]
  34. Nurmikko V., Laaksonen S. 1961; Biosynthesis and excretion of vitamin B6 by Escherichia coli during the exponential phase of growth. Acta chem. fenn. B 34:7
    [Google Scholar]
  35. Pardee A. B. 1955; Effect of energy supply on enzyme induction by pyrimidine requiring mutants of Escherichia ccli. J. Bact 69:233
    [Google Scholar]
  36. Pardee A. B. 1957; An inducible mechanism for the accumulation of melibiose in Escherichia coli. J. Bact 73:376
    [Google Scholar]
  37. Pardee A. B. 1959; The control of enzyme activity. The Enzymes (second edition) I, 681 Boyer H., Lardy P. D., Myrbäck K. New York: Academic Press;
    [Google Scholar]
  38. Pardee A. B., Prestidge L. S. 1959; On the nature of the repressor of β-galactosidase synthesis in Escherichia coli. Biochim. biophys. Acta 36:545
    [Google Scholar]
  39. Pardee A. B., Shore V. G., Prestidge L. S. 1956; Incorporation of azatryptophan into proteins of bacteria and bacteriophage. Biochim. biophys. Acta 21:406
    [Google Scholar]
  40. Peel J. L. 1958; The separation of flavins by paper electrophoresis and its application to the examination of the flavin contents of micro-organisms. Biochem. J 69:403
    [Google Scholar]
  41. Plaut G. W. E. 1954; Biosynthesis of riboflavin I. Incorporation of C14-labelled compounds into rings B and C. J. biol. Chem 208:513
    [Google Scholar]
  42. Plaut G. W. E. 1960; Studies on the stoichiometry of the enzymic conversion of 6,7-dimethyl-8-ribityllumazine to riboflavin. J. biol. Chem 235:PC41
    [Google Scholar]
  43. Prestidge L. S., Pardee A. B. 1957; Induction of bacterial lysis by penicillin. J. Bact 74:48
    [Google Scholar]
  44. Roberts R. B., Abelson P. H., Cowie D. B., Bolton E. T., Britten R. J. 1955; Studies of biosynthesis in Escherichia coli. Publ. Carneg. Instn (Wash.) 607:1
    [Google Scholar]
  45. Roe J. H., Epstein J. H., Goldstein N. P. 1949; A photometric method for the determination of inulin in plasma and urine. J. biol. Chem 178:839
    [Google Scholar]
  46. Rushizky G., Riley M., Prestidge L. S., Pardee A. B. 1960; Inactivation of enzyme formation by ultraviolet light I. Action spectra and the size of the sensitive unit. Biochim. biophys. Acta 45:70
    [Google Scholar]
  47. Schrecker A. W., Kornberg A. 1950; Reversible enzymatic synthesis of flavin-adenine dinucleotide. J. biol. Chem 182:795
    [Google Scholar]
  48. Shiota T. 1959; Enzymic synthesis of folic acid-like compounds by cell-free extracts of Lactobacillus arabinosus. Arch. Biochem. Biophys 80:155
    [Google Scholar]
  49. Singer T. P., Kearney E. B., Massey V. 1956; Observations on the flavin moiety of succinic dehydrogenase. Arch. Biochem. Biophys 60:255
    [Google Scholar]
  50. Smith C. G., Smith G. A., Papadoupoulou Z. 1961; Effects of surface active agents on the biosynthesis of riboflavin by Ashbya gossypii. Biochim,. biophys. Acta 47:344
    [Google Scholar]
  51. Snoswell A. M. 1957; Flavokinase of Lactobacillus arabinosus 17.5. Aust. J. exp. Biol, med. Sci 35:427
    [Google Scholar]
  52. Snell E. E. 1950; Microbiological methods in vitamin research. Vitamin Methods I:327 György P. New York: Academic Press;
    [Google Scholar]
  53. Thompson R. C. 1942; Synthesis of B vitamins by bacteria in pure culture. Univ. Tex. Publ. No. 423787
    [Google Scholar]
  54. Troll W., Cannan R. K. 1953; A modified photometric ninhydrin method for the analysis of amino and imino acids. J. biol. Chem 200:803
    [Google Scholar]
  55. van Lanen J. M. 1948; Vitamins in microorganisms—distribution and quantitative syntheses. Vitam. & Horm 6:163
    [Google Scholar]
  56. Warringa M. G. P. J., Giuditta A. 1958; Studies on succinic dehydrogenase, IX. Characterization of the enzyme from Micrococcus lactilyticus. J. biol. Chem 230:111
    [Google Scholar]
  57. Webb M. 1954; The effects of folic acid analogues on the growth and cell division of micro-organisms. The Chemistry and Biology of Pteridines, Ciba Foundation Symposium253 Boston: Little, Brown and Co.;
    [Google Scholar]
  58. Weiss B., Srinivasan P. R. 1959; The biosynthesis of p-aminobenzoic acid. Proc. nat. Acad. Sci., Wash 45:1491
    [Google Scholar]
  59. Wilson A. C., Pardee A. B. 1962; Comparative aspects of metabolic control. Comparative Biochemistry V. Florkin M., Mason H. S. New York: Academic Press.; (In the Press.)
    [Google Scholar]
  60. Wisseman C. L., Smadel J. E., Hahn F. E., Hopps H. E. 1954; Mode of action of chloramphenicol, I. Action of chloramphenicol on assimilation of ammonia and on synthesis of proteins and nucleic acids. J. Bact 67:662
    [Google Scholar]
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