title: Biosynthesis of vitamin B-12 in anaerobic bacteria. Experiments with Eubacterium limosum on the origin of the amide groups of the corrin ring and of N-3 of the 5,6-dimethylbenzimidazole part.
abstract: The pathway of vitamin B-12 biosynthesis in anaerobic bacteria differs in several respects from the pathway found in aerobic or aerotolerant microorganisms. The aim of this investigation was to elucidate the formation of the 5,6-dimethylbenzimidazole part and the amide groups of vitamin B-12 in anaerobic bacteria. [15N]Ammonium chloride or L-[amido-15N]glutamine or a mixture of [15N]ammonium sulfate and [15N]glycine was added to fermentations with Eubacterium limosum. The vitamin B-12 isolated from these fermentations was methylated and degraded to cobinamide and 1,5,6-trimethylbenzimidazole. The amide groups of cobinamide were hydrolyzed and the amide nitrogen of the side chains a, b, c, d, e and g trapped as benzamide. The 15N incorporation was determined by mass spectroscopy. Thus in the experiment with [15N]ammonium chloride the benzamide and the 1,5,6-trimethylbenzimidazole contained 9.6% 15N, whereas in the experiment with L-[amido-15N]glutamine 37.5% of the molecules were 15N labeled. The 1H-NMR spectrum of 1,5,6-trimethylbenzimidazole revealed that the 15N from the ammonium salts and from glutamine was incorporated into N-3 of the 5,6-dimethylbenzimidazole moiety of vitamin B-12. With a mixture of [15N]ammonium sulfate and [15N]glycine both nitrogens of 5,6-dimethylbenzimidazole became 15N-labeled. These experiments demonstrate that in E. limosum the amide nitrogen of glutamine is not only the precursor of the six amide groups of the corrin ring, but also of N-3 of the 5,6-dimethylbenzimidazole moiety of vitamin B-12.
authors.affiliation: Institut für Biologische Chemie, Universität Hohenheim, Stuttgart, Federal Republic of Germany.
authors.forename: J R
authors.initials: JR
authors.lastname: Vogt
authors.forename: P
authors.initials: P
authors.lastname: Renz
mesh_terms: D000577:Amides / Q000096:biosynthesis*; D000693:Anaerobiosis; D001562:Benzimidazoles / Q000096:biosynthesis*; D045728:Corrinoids; D005051:Eubacterium / Q000378:metabolism*; D005285:Fermentation; D006868:Hydrolysis; D009682:Magnetic Resonance Spectroscopy; D008745:Methylation; D014805:Vitamin B 12 / Q000096:biosynthesis*
chemical_list: D000577:Amides; D001562:Benzimidazoles; D045728:Corrinoids; C015158:5,6-dimethylbenzimidazole; D014805:Vitamin B 12