Biosynthesis of porphyrins and related macrocycles, part 43. Isolation and characterization of intermediates of coenzyme B12 biosynthesis, a cobyrinic acid triamide, the a,c-diamide and their Co-(5′-deoxy-5′-adenosyl) derivatives, from Propionibacterium shermanii

AbstractBackground: Vitamin B12 is synthesized by many different organisms, for example Pseudomonas denitrificans (aerobic) and Propionibacterium shermanii (‘microaerophilic’, or essentially anaerobic). The biosynthetic pathways in these two organisms show strong similarities but also some differences. There have been conflicting reports on where differences between these two organisms lie in the stages beyond the formation of the corrin macrocycle. Characterization of intermediates in the pathway will help resolve these conflicts.Results: A single cobyrinic acid diamide and a single triamide have been isolated from Pr. shermanii.The diamide was shown to be the a,c-isomer. The triamide is not the a,c,g-isomer but it is indistinguishable from the single triamide isolated by other workers from Ps. denitrificans. The Co-(5′-deoxy-5′-adenosyl) derivative of the a,c-diamide was also isolated and fully characterized and the deoxyadenosyl derivative of the foregoing triamide has been shown to be present in the cells.Conclusions: Our results support a unique pathway in Pr. shermanii proceeding from cobyrinic acid towards coenzyme B12, at least as far as the adenosylated triamide intermediate. No evidence was found for multiple alternative pathways. The order of amidations of the carboxyl side-chains of cobyrinic acid up to the triamide stage is the samee in Pr. shermanii and Ps. denitrificans