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

doi:10.1111/j.1348-0421.1993.tb01732.x

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 journal › Article › peer-review

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 language	English
Pages (from-to)	975-978
Number of pages	4
Journal	MICROBIOLOGY and IMMUNOLOGY
Volume	37
Issue number	12
DOIs	https://doi.org/10.1111/j.1348-0421.1993.tb01732.x
Publication status	Published - Dec 1 1993

Keywords

Hemolysin
Hydrophobic chromatography
Vibrio vulnificus

ASJC Scopus subject areas

Microbiology
Immunology
Virology

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10.1111/j.1348-0421.1993.tb01732.x

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

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.",

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AU - Oh, Eun gyoung

AU - Tamanoi, Yumi

AU - Toyoda, Atsuko

AU - Usui, Kaori

AU - Miyoshi, Shin ichi

AU - Chang, Dong Suck

AU - Shinoda, Sumio

PY - 1993/12/1

Y1 - 1993/12/1

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.

AB - 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.

KW - Hemolysin

KW - Hydrophobic chromatography

KW - Vibrio vulnificus

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

Abstract

Keywords

ASJC Scopus subject areas

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