Simple Purification Method for a Vibrio vulnificus Hemolysin by a Hydrophobic Column Chromatography in the Presence of a Detergent

Eun gyoung Oh, Yumi Tamanoi, Atsuko Toyoda, Kaori Usui, Shin ichi Miyoshi, Dong Suck Chang, Sumio Shinoda

Research output: Contribution to journalArticle

15 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
DOIs
Publication statusPublished - Dec 1 1993

Keywords

  • Hemolysin
  • Hydrophobic chromatography
  • Vibrio vulnificus

ASJC Scopus subject areas

  • Microbiology
  • Immunology
  • Virology

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