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Purification of α-hemolysin from an overproducing E. coli strain

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Summary

The genetic determinant of the α-hemolysin encoded by plasmid pHly152 has been cloned in both orientations in plasmid pBR322 giving rise to plasmids pSU157 and pSU158.

E. coli strains carrying either of these recombinant Hly plasmids produced about 20 times more hemolysin activity than the parental plasmid pHly152, when grown in minimal medium supplemented with hemoglobin. Thus high hemolytic activity is not lethal to the cells, contrary to previous assumptions.

α-hemolysin was purified from culture supernatants of strain SU100 (pSU157) by ammonium sulfate precipitation and gel filtration in Sephacryl S-200 in the presence of 6 M urea. When purified α-hemolysin preparations were subjected to electrophoretic analysis in denaturing conditions, a single 107 kdal polypeptide was observed. This probably corresponds to the α-hemolysin protein, since an isogenic E. coli strain carrying plasmid pSU161, an Hly- mutant derivative of pSU157, did not synthesize the 107 kdal polypeptide.

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Authors and Affiliations

  1. Departmento de Bioquimica, Facultad de Medicina, Universidad de Santander, Santander, Spain

    M. I. Gonzalez-Carreró, J. C. Zabala & F. de la Cruz

  2. Hospital Nacional Marques de Valdecilla, Santander, Spain

    J. M. Ortiz

Authors
  1. M. I. Gonzalez-Carreró
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  2. J. C. Zabala
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  3. F. de la Cruz
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  4. J. M. Ortiz
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Communicated by A. Böck

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Gonzalez-Carreró, M.I., Zabala, J.C., de la Cruz, F. et al. Purification of α-hemolysin from an overproducing E. coli strain. Molec Gen Genet 199, 106–110 (1985). https://doi.org/10.1007/BF00327518

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  • DOI: https://doi.org/10.1007/BF00327518

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