Novel Compounds From Methanogens: Characterization of Component B of the Methylreductase System and Mobile Factor
An enzymic assay for the detection and quantitation of component B based on cell-free extract of Methanobacterium thermoautotrophicum stripped of small molecular weight cofactors by anaerobic column chromatography was developed. Component B was purified to homogeneity from boiled cell extract of M. thermoautotrophicum by anaerobic chromatography on Sephadex DEAE-A25, Sephadex G10, and Sephadex G25-F columns. Component B was determined to have a molecular weight of 1,044 by fast atom bombardment mass spectrometry, and a molecular formula of C(,32)H(,47)N(,3)O(,26)P(,2)Na(,4) was proposed for the sodium salt of the compound. A primary amine was present in component B, but no amino acid was detected. Component B was determined to have a monoester phosphate and a diester phosphate, and one mol of aldopentose per mol of component B. The UV absorbance spectrum revealed the presence of two absorbance peaks in component B: one at 267 nm and the other at 233 nm. It was concluded that component B contained a nucleotide moiety.
University of Illinois - Urbana-Champaign
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University of Illinois at Urbana-Champaign