cobyric-acid and cobinamide

Vitamin B12 plays a key role in many metabolic processes occurring in all mammals. Over the years its biological role has been extensively studied generating a lot of interest in the chemistry of this vital molecule. This established a variety of new methodologies for the synthesis and analysis of new cobalamin derivatives as well as creative purification techniques. This tutorial review summarizes all the advancements made in this area, providing a deeper insight into vitamin B12 chemistry.

Topics: Cobalt; Cobamides; Lactams; Lactones; Vitamin B 12

The existence of a pathway for salvaging the coenzyme B(12) precursor dicyanocobinamide (Cbi) from the environment was established by genetic and biochemical means. The pathway requires the function of a previously unidentified amidohydrolase enzyme that converts adenosylcobinamide to adenosylcobyric acid, a bona fide intermediate of the de novo coenzyme B(12) biosynthetic route. The cbiZ gene of the methanogenic archaeon Methanosarcina mazei strain Göl was cloned, was overproduced in Escherichia coli, and the recombinant protein was isolated to homogeneity. HPLC, UV-visible spectroscopy, MS, and bioassay data established adenosylcobyric as the corrinoid product of the CbiZ-catalyzed reaction. Inactivation of the cbiZ gene in the extremely halophilic archaeon Halobacterium sp. strain NRC-1 blocked the ability of this archaeon to salvage Cbi. cbiZ function restored Cbi salvaging in a strain of the bacterium Salmonella enterica, whose Cbi-salvaging pathway was blocked. The salvaging of Cbi through the CbiZ enzyme appears to be an archaeal strategy because all of the genomes of B(12)-producing archaea have a cbiZ ortholog. Reasons for the evolution of two distinct pathways for Cbi salvaging in prokaryotes are discussed.

Topics: Aminohydrolases; Cobamides; Genes, Archaeal; Genetic Complementation Test; Halobacterium; Methanosarcina; Molecular Sequence Data; Mutation; Recombinant Proteins; Salmonella enterica; Species Specificity; Vitamin B 12

The cyanoaquo and aquocyano stereoisomers of several putative vitamin B12 precursors are reversibly formed and can be separated using analytical high-performance liquid chromatographic methods. The behavior of these stereoisomers varies somewhat depending on the type of column used and the chromatographic conditions employed. Both reversed-phase and ion-exchange columns can be used to observe the reversible formation and separation of the stereoisomers of (H2O,CN)cobyric acid, cobinamide and the cobinic acid pentaamide-1, -2 and -3 structural isomers. The greatest differences in retention times are seen when the pH of the eluting buffer is less than 4.0 and the buffer contains no KCN.

Topics: Chromatography, High Pressure Liquid; Chromatography, Thin Layer; Cobamides; Corrinoids; Hydrogen-Ion Concentration; Spectrophotometry, Ultraviolet; Stereoisomerism; Vitamin B 12

The preparation of the diaquo- forms of the incomplete corrinoids, cobyric acid, cobinamide, and three isomeric cobinic acid pentaamides, from the corresponding aquocyanocorrinoids has been accomplished. The compounds were characterized by electronic absorption spectra, pka values, and HPLC/TLC behavior. Upon titration, the acids exhibited three pKa values, but only the pKa corresponding to the carboxylic acid involved the transfer of a full equivalent of protons. In addition, the progress of diaquo- to dihydroxo-titration, and the spectrum of the final products indicated a complex process. These diaquocorrinoids appear to be unstable, and should be stored in solution, frozen, under an inert atmosphere.

Topics: Chromatography, High Pressure Liquid; Cobamides; Isomerism; Kinetics; Molecular Structure; Oxidation-Reduction; Vitamin B 12