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Purification and analysis of cobamides of Methanobacterium bryantii by high-performance liquid chromatography

✍ Scribed by William B. Whitman; Ralph S. Wolfe

Publisher
Elsevier Science
Year
1984
Tongue
English
Weight
379 KB
Volume
137
Category
Article
ISSN
0003-2697

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✦ Synopsis

Cobamides from Methanobacterium brvantii were purified by isocratic reverse-phase highperformance liquid chromatography at neutral pH in readily extractable buffers. A gradient adaptation of this chromatography system on C-18 PBondapak columns in buffers containing aqueous methanol and 100 mM LiCl readily separated Coa-(5-hydroxybenzimidazoyl)-Coflcyanocobamide, Cool-(5-hydroxybenzimidazoyl)-Cog-adenosylcobamide. cyanocobalamin, adenosylcobalamin, and methylcobalamin. Aquacobalamin was not resolved from cyanocobalamin by this procedure.

The gradient procedure was also useful in following the fate of tritiated cyanocobalamin in cell-free extracts of M. bryznfii.

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