1887

Abstract

SUMMARY: type A (13124) was grown in bottles (static culture) and in fermenters (batch and continuous culture) in a proteose peptone medium supplemented with phosphate, cysteine, vitamins and different carbohydrates. The lag phase was shorter and the growth rate higher when the medium was pre-reduced. The final yields of bacteria, phospholipase C, and theta-haemolysin were also significantly higher in the pre-reduced medium. The optimum pH for maximum bacterial yield was 7·0 and the optimum temperature for growth was 37°C. The formation of phospholipase C was optimum between pH 6·0 and 7·0 and for theta-haemolysin between pH 7·0 and 7·5. The optimum temperaure for synthesis of both phospholipase C and theta-haemolysin was 37°C. Culture under controlled conditions gave more reproducible production of these two proteins than experiments in bottles. The cultures reached the stationary phase before phospholipase C activity started to decline, while theta-haemolysin activity was stable. The strain was also grown in continuous culture at a dilution rate of 0·4 h, at pH 7·0 and 37°C, with a yield of 1·2 mg dry wt/ml in the complex medium supplemented with glucose (5 g/l). After the continuous feed was started the activity of phospholipase C declined very rapidly to zero, while theta-haemolysin continued to be produced at a constant level. Thus continuous culture offered no benefit over batch culture for reproducible laboratory-scale production of phospholipase C.

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1974-09-01
2024-03-29
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References

  1. Arvidson S., Holme T., Wadström T. 1970; Formation of bacteriolytic enzymes in batch and continuous culture of Staphylococcus aureus . Journal of Bacteriology 104:227–233
    [Google Scholar]
  2. Arvidson S., Holme T., Wadström T. 1971; Influence of cultivation conditions on the production of extracellular proteins by Staphylococcus aureus . Acta pathologica et microbiologica scandinavica 79B:399–405
    [Google Scholar]
  3. Chou G. 1971; Medium for toxin production of Clostridium perfringens in continuous culture. Applied Microbiology 21:794–798
    [Google Scholar]
  4. Gale E. F., van Heyningen W. E. 1942; The effect of the pH and presence of glucose during growth on the production of α and θ toxins and hyaluronidase by Clostridium welchii . Biochemical Journal 36:624–630
    [Google Scholar]
  5. Goldbarg J. A., Rutenberg A. M. 1965; The colorimetric determination of leucine aminopeptidase in urine and serum of normal subjects and patients with cancer and other diseases. Cancer 11:283–287
    [Google Scholar]
  6. Hauschild A. H. W. 1966; Selective effect of pH on the production of extracellular protein by Clostridium perfringens type D. Journal of Bacteriology 92:800–801
    [Google Scholar]
  7. Hauschild A. H. W., Pivnick H. 1965; Effect of carbohydrates on toxinogenesis by Clostridium perfringens type D. Canadian Journal of Microbiology 11:15–22
    [Google Scholar]
  8. Holdeman L. V., Moore W. E. C. 1972 Anaerobe Laboratory Manual Blacksburg, Virginia: Virginia Polytechnic Institute and State University;
    [Google Scholar]
  9. Ispolatovskaya M. V. 1971; Type A Clostridium perfringens toxin. In Microbial Toxins - Bacterial Protein Toxins vol. 2A pp. 109–158 Edited by Kadis S., Montie T. C., Ajl S. J. New York and London: Academic Press;
    [Google Scholar]
  10. Jacob H.-E. 1970; Redox potential. In Methods in Microbiology vol. 2 pp. 91–123 Edited by Norris J. R., Ribbons D. W. London and New York: Academic Press;
    [Google Scholar]
  11. Jayko L. G., Lichstein H. C. 1959; Nutritional factors concerned with growth and lecithinase production by Clostridium perfringens . Journal of Infectious Diseases 104:142–151
    [Google Scholar]
  12. Lowry O. H., Rosebrough N. J., Farr A. L., Randall R. J. 1951; Protein measurement by the Folin phenol reagent. Journal of Biological Chemistry 193:265–275
    [Google Scholar]
  13. Möllby R., Nord C.-E., Wadstrom T. 1973; Biological activities contaminating preparations of phospholipase C (α-toxin) from Clostridium perfringens . Toxicon 11:139–147
    [Google Scholar]
  14. Möllby R., Wadstrom T. 1973; Purification of phospholipase C (alpha toxin) from Clostridium perfringens . Biochimica et biophysica acta 321:569–584
    [Google Scholar]
  15. Moore W. E. C., Cato E. P., Holdeman L. V. 1969; Anaerobic bacteria of the gastrointestinal flora and their occurrence in clinical infections. Journal of Infectious Diseases 119:641–649
    [Google Scholar]
  16. Murata R., Soda S., Yamamoto A., Ito A. 1968; Further investigations on the influence of inorganic cations on growth and toxin production by Clostridium perfringens pb6k . Japanese Journal of Medical Science and Biology 21:55–70
    [Google Scholar]
  17. Murata R., Soda S., Yamamoto A., Sato H., Ito A. 1969; The effect of zinc on the production of various toxins of Clostridium perfringens . Japanese Journal of Medical Science and Biology 22:133–148
    [Google Scholar]
  18. Murata R., Yamamoto A. 1964; A synthetic medium for production of alpha toxin by Clostridium perfringens . Japanese Journal of Medical Science and Biology 17:211–213
    [Google Scholar]
  19. Murata R., Yamada T., Kameyama S. 1956; Production of alpha toxin of Clostridium perfringens. I. Preparation of the reproducible peptone medium for the production of the toxin of high potency. Japanese Journal of Medical Science and Biology 9:81–91
    [Google Scholar]
  20. Murata R., Yamamoto A., Soda S., Ito A. 1965; Nutritional requirements of Clostridium perfringens pb6k for alpha toxin production. Japanese Journal of Medical Science and Biology 18:189–202
    [Google Scholar]
  21. Murphey W. H., Barnaby C., Lin F. J., Kaplan N. O. 1967; Malate dehydrogenases. II. Purification and properties of Bacillus subtilis, Bacillus stearothermophilus and Escherichia coli malate dehydrogenases. Journal of Biological Chemistry 242:1548–1559
    [Google Scholar]
  22. Nakamura M., Cook J. A., Cross W. R. 1968; Lecithinase production by Clostridium perfringens in chemically defined media. Applied Microbiology 16:1420–1421
    [Google Scholar]
  23. Nakamura M., Schulze J. A., Cross W. R. 1969; Factors affecting lecithinase activity and production in Clostridium welchii . Journal of Hygiene 67:153–162
    [Google Scholar]
  24. Pivnick H., Habeeb A. F. S. A., Gorenstein B., Stuart P. F., Hauschild A. H. W. 1964; Effect of pH on toxinogenesis by Clostridium perfringens type C. Canadian Journal of Microbiology 10:329–344
    [Google Scholar]
  25. Ricica J. 1971; Continuous cultivation of micro-organisms. Folia microbiologica (Praha) 16:389–415
    [Google Scholar]
  26. Sargeant K. 1968; Improvement of yields in anaerobic cultures. Chemistry and Industry 1968:85–88
    [Google Scholar]
  27. Schulze J. A., Nakamura M. 1968; Lecithinase activity of mutant and wild strains of Clostridium perfringens . Applied Microbiology 16:1619–1620
    [Google Scholar]
  28. Shemanova G. F., Gorshkova V. I., Borisova O. K., Shakhanina K. L. 1970; Study of the reciprocal relationship of the exo-antigens of Clostridium perfringens type A in the culture medium and the bacterial cells during cultivation. Folia microbiologica (Praha) 15:23–33
    [Google Scholar]
  29. Shemanova G. F., Vlasova E. V., Shamraeva S. A. 1964; Action of the proteinases of Clostridium oedematiens and Clostridium perfringens on homologous and heterologous toxins. Bulletin of Experimental Biology and Medicine 57:457–460
    [Google Scholar]
  30. Smith L. D. S., Holdeman L. V. 1968 The Pathogenic Anaerobic Bacteria Springfield, Illinois: Charles C. Thomas;
    [Google Scholar]
  31. Soda S., Sato H., Murata R. 1969; The effect of calcium on the production of various toxins of Clostridium perfringens . Japanese Journal of Medical Science and Biology 22:175–179
    [Google Scholar]
  32. Tabatabai L. B., Walker H. W. 1970; Oxidation reduction potential and growth of Clostridium perfringens and Pseudomonas fluorescens . Applied Microbiology 20:441–446
    [Google Scholar]
  33. Vinet G., Fredette V. 1970; £tude de la toxinogenese de Plectridium tetani . Canadian Journal of Microbiology 16:1067–1070
    [Google Scholar]
  34. Willis A. T. 1969 Clostridia of Wound Infections London: Butterworths;
    [Google Scholar]
  35. Wretlind B., Heden L., Wadstrom T. 1973; Formation of extracellular haemolysin by Aeromonas hydrophila in relation to protease and staphylolytic enzyme. Journal of General Microbiology 78:57–65
    [Google Scholar]
  36. Zacharias B., Bjorklund M. 1968; Continuous production of Clostridium tetani toxin. Applied Microbiology 16:69–72
    [Google Scholar]
  37. Zwaal R. F. A., Roelofsen B., Comfurius P., van Deenen L. L. M. 1971; Complete purification and some properties of phospholipase C from Bacillus cereus . Biochimica et biophysica acta 233:474–479
    [Google Scholar]
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