corrin and cobyrinic-acid

It has been known for the past 20 years that two pathways exist in nature for the de novo biosynthesis of the coenzyme form of vitamin B12, adenosylcobalamin, representing aerobic and anaerobic routes. In contrast to the aerobic pathway, the anaerobic route has remained enigmatic because many of its intermediates have proven technically challenging to isolate, because of their inherent instability. However, by studying the anaerobic cobalamin biosynthetic pathway in Bacillus megaterium and using homologously overproduced enzymes, it has been possible to isolate all of the intermediates between uroporphyrinogen III and cobyrinic acid. Consequently, it has been possible to detail the activities of purified cobinamide biosynthesis (Cbi) proteins CbiF, CbiG, CbiD, CbiJ, CbiET, and CbiC, as well as show the direct in vitro conversion of 5-aminolevulinic acid into cobyrinic acid using a mixture of 14 purified enzymes. This approach has resulted in the isolation of the long sought intermediates, cobalt-precorrin-6A and -6B and cobalt-precorrin-8. EPR, in particular, has proven an effective technique in following these transformations with the cobalt(II) paramagnetic electron in the dyz orbital, rather than the typical dz2. This result has allowed us to speculate that the metal ion plays an unexpected role in assisting the interconversion of pathway intermediates. By determining a function for all of the pathway enzymes, we complete the tool set for cobalamin biosynthesis and pave the way for not only enhancing cobalamin production, but also design of cobalamin derivatives through their combinatorial use and modification.

Topics: Anaerobiosis; Bacillus megaterium; Bacterial Proteins; Biosynthetic Pathways; Corrinoids; Electron Spin Resonance Spectroscopy; Models, Chemical; Molecular Structure; Nuclear Magnetic Resonance, Biomolecular; Vitamin B 12

The synthesis of cobyrinic acid derivatives by reduction of dehydrocobyrinates is largely unexplored. It is, however, a rational path to B(12) analogues that lack specific substituents of the corrin moiety of natural B(12) derivatives. The partial syntheses of four epimeric 7-decarboxymethyl-cobyrinates is described, which is achieved by reduction of Δ7-dehydro-7-de[carboxymethyl]-cobyrinate with zinc or with the 'prebiotic' reducing agent formic acid. A direct and remarkably efficient route was found to 7-decarboxymethyl-cobyrinates, which are cobyrinic acid derivatives in which the c-side chain at ring B of vitamin B(12) is missing. The structures of the hexamethyl-7-decarboxymethyl-cobyrinates were characterized and the stereochemical and conformational properties at their newly saturated ring B were analyzed. The stereochemical outcome of the reduction was found to depend strongly on the reaction conditions. In 7-decarboxymethyl-cobyrinates, both peripheral carbon centres of ring B carry a hydrogen atom, and the characteristic quaternary carbon centre at C7 of the cobyrinic acid moiety of vitamin B(12) is lacking. The still highly substituted 7-decarboxymethyl-cobyrinates are readily dehydrogenated in the presence of dioxygen, furnishing 7-de[carboxymethyl]-Δ(7)-dehydro-cobyrinate as the common, unsaturated oxidation product. The noted stability of vitamin B(12) and of other Co(III)-cobyrinates in the presence of air is a consequence of their highly substituted corrin macrocycle, a finding of interest in the context of chemical rationalizations of the B(12) structure.

Topics: Carbon; Corrinoids; Molecular Structure; Vitamin B 12