Simple Purification Method for aVibrio vulnificusHemolysin by a Hydrophobic Column Chromatography in the Presence of a Detergent

Autor:	Yumi Tamanoi, Shin Ichi Miyoshi, Eun gyoung Oh, Dong Suck Chang, Kaori Usui, Atsuko Toyoda, Sumio Shinoda
Rok vydání:	1993
Předmět:	Chromatography biology Toxin Elution Detergents Immunology Hemolysin Vibrio vulnificus biology.organism_classification medicine.disease_cause Microbiology Hemolysin Proteins Column chromatography Biochemistry Vibrionaceae Chaps Virology medicine Polyacrylamide gel electrophoresis Vibrio
Zdroj:	Microbiology and Immunology. 37:975-978
ISSN:	0385-5600
DOI:	10.1111/j.1348-0421.1993.tb01732.x
Popis:	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.
Databáze:	OpenAIRE
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