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. 1987 Feb;25(2):222–225. doi: 10.1128/jcm.25.2.222-225.1987

Production and purification of Streptococcus pneumoniae hemolysin (pneumolysin).

K Kanclerski, R Möllby
PMCID: PMC265871  PMID: 3818918

Abstract

Pneumolysin was found to be produced by 112 of 113 clinical isolates of Streptococcus pneumoniae and to be an intracellular hemolysin. A 10-liter-scale fermentor production and purification procedure was developed for this hemolysin. The culture was concentrated by filtration 10 times before centrifugation. The cellular content was purified by ion-exchange chromatography, covalent thiopropyl gel chromatography, and gel filtration. One batch operation resulted in 6 mg of highly purified pneumolysin, with a yield of 66% and a specific activity of 1,400,000 hemolytic units per mg. The pneumolysin had a molecular weight of 53,000 and an isoelectric point of 5.2. The purification method developed will be of value in future studies on this hemolysin.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Cohen B., Halbert S. P., Perkins M. E. Pneumococcal Hemolysin: The Preparation of Concentrates, and Their Action on Red Cells. J Bacteriol. 1942 May;43(5):607–627. doi: 10.1128/jb.43.5.607-627.1942. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Ferrante A., Rowan-Kelly B., Paton J. C. Inhibition of in vitro human lymphocyte response by the pneumococcal toxin pneumolysin. Infect Immun. 1984 Nov;46(2):585–589. doi: 10.1128/iai.46.2.585-589.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Johnson M. K., Allen J. H. Ocular toxin of the pneumococcus. Am J Ophthalmol. 1971 Jul 30;72(1):175–180. doi: 10.1016/0002-9394(71)91610-2. [DOI] [PubMed] [Google Scholar]
  4. Johnson M. K., Boese-Marrazzo D., Pierce W. A., Jr Effects of pneumolysin on human polymorphonuclear leukocytes and platelets. Infect Immun. 1981 Oct;34(1):171–176. doi: 10.1128/iai.34.1.171-176.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Johnson M. K., Hamon D., Drew G. K. Isolation and characterization of pneumolysin-negative mutants of Streptococcus pneumoniae. Infect Immun. 1982 Aug;37(2):837–839. doi: 10.1128/iai.37.2.837-839.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Johnson M. K., Knight R. J., Drew G. K. The hydrophobic character of thiol-activated cytolysins. Biochem J. 1982 Dec 1;207(3):557–560. doi: 10.1042/bj2070557. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Johnson M. K. Properties of purified pneumococcal hemolysin. Infect Immun. 1972 Nov;6(5):755–760. doi: 10.1128/iai.6.5.755-760.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Kalin M., Kanclerski K., Granström M., Möllby R. Diagnosis of pneumococcal pneumonia by enzyme-linked immunosorbent assay of antibodies to pneumococcal hemolysin (pneumolysin). J Clin Microbiol. 1987 Feb;25(2):226–229. doi: 10.1128/jcm.25.2.226-229.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Kallings I., Bengtsson S., Christensen P., Holm S. E., Lind L., Kalin M. Antibiotic sensitivity of Haemophilus influenzae, Streptococcus pneumoniae, Streptococcus pyogenes and Branhamella catarrhalis isolated from upper respiratory tract infections in Sweden. Scand J Infect Dis Suppl. 1983;39:100–105. [PubMed] [Google Scholar]
  10. Kreger A. S., Bernheimer A. W. Physical behavior of pneumolysin. J Bacteriol. 1969 Apr;98(1):306–307. doi: 10.1128/jb.98.1.306-307.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Lorian V., Waluschka A., Popoola B. Pneumococcal beta hemolysin produced under the effect of antibiotics. Appl Microbiol. 1973 Feb;25(2):290–294. doi: 10.1128/am.25.2.290-294.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Paton J. C., Lock R. A., Hansman D. J. Effect of immunization with pneumolysin on survival time of mice challenged with Streptococcus pneumoniae. Infect Immun. 1983 May;40(2):548–552. doi: 10.1128/iai.40.2.548-552.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Paton J. C., Rowan-Kelly B., Ferrante A. Activation of human complement by the pneumococcal toxin pneumolysin. Infect Immun. 1984 Mar;43(3):1085–1087. doi: 10.1128/iai.43.3.1085-1087.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Shumway C. N., Klebanoff S. J. Purification of pneumolysin. Infect Immun. 1971 Oct;4(4):388–392. doi: 10.1128/iai.4.4.388-392.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]

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