Simple purification method for a Vibrio vulnificus hemolysin by a hydrophobic column chromatography in the presence of a detergent

E. G. Oh, Y. Tamanoi, A. Toyoda, K. Usui, Shin-ichi Miyoshi, D. S. Chang, S. Shinoda

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

A Vibrio vulnificus hemolysin (VVH) was purified by two steps of hydrophobic column chromatography on Phenyl-Sepharose HP. The first chromatography was carried out at pH 6.0. In this pH condition, VVH efficiently bound to the column, but the hemolysin fraction eluted was accompanied with colored substance(s). To eliminate this colored substance, the second chromatography was carried out at pH 9.8 in the presence of 1% 3- [(3-cholamidopropyl) dimethylammonio]-1-propanesulfonate (CHAPS), a zwitterionic detergent. Homogeneity of the hemolysin thus obtained was shown by polyacrylamide gel electrophoresis. The specific activity increased 33,600 times and the yield was 35%. The method is simple and useful to supply enough VVH for study of the role of the hemolysin in the infection by V. vulnificus or on the mechanism of action of the hemolysin.

Original languageEnglish
Pages (from-to)975-978
Number of pages4
JournalMicrobiology and Immunology
Volume37
Issue number12
Publication statusPublished - 1993

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Vibrio vulnificus
Hemolysin Proteins
Detergents
Chromatography
Polyacrylamide Gel Electrophoresis
Vibrio hemolysin

ASJC Scopus subject areas

  • Immunology and Microbiology(all)
  • Microbiology
  • Microbiology (medical)

Cite this

Simple purification method for a Vibrio vulnificus hemolysin by a hydrophobic column chromatography in the presence of a detergent. / Oh, E. G.; Tamanoi, Y.; Toyoda, A.; Usui, K.; Miyoshi, Shin-ichi; Chang, D. S.; Shinoda, S.

In: Microbiology and Immunology, Vol. 37, No. 12, 1993, p. 975-978.

Research output: Contribution to journalArticle

Oh, E. G. ; Tamanoi, Y. ; Toyoda, A. ; Usui, K. ; Miyoshi, Shin-ichi ; Chang, D. S. ; Shinoda, S. / Simple purification method for a Vibrio vulnificus hemolysin by a hydrophobic column chromatography in the presence of a detergent. In: Microbiology and Immunology. 1993 ; Vol. 37, No. 12. pp. 975-978.
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AU - Tamanoi, Y.

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AU - Usui, K.

AU - Miyoshi, Shin-ichi

AU - Chang, D. S.

AU - Shinoda, S.

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N2 - A Vibrio vulnificus hemolysin (VVH) was purified by two steps of hydrophobic column chromatography on Phenyl-Sepharose HP. The first chromatography was carried out at pH 6.0. In this pH condition, VVH efficiently bound to the column, but the hemolysin fraction eluted was accompanied with colored substance(s). To eliminate this colored substance, the second chromatography was carried out at pH 9.8 in the presence of 1% 3- [(3-cholamidopropyl) dimethylammonio]-1-propanesulfonate (CHAPS), a zwitterionic detergent. Homogeneity of the hemolysin thus obtained was shown by polyacrylamide gel electrophoresis. The specific activity increased 33,600 times and the yield was 35%. The method is simple and useful to supply enough VVH for study of the role of the hemolysin in the infection by V. vulnificus or on the mechanism of action of the hemolysin.

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